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Administrative data

Description of key information

Male and female Wistar rats (20 animals per sex and dose group) received 22 daily oral (gavage) doses of 0 or 1000 mg test item per kg bw over a period of 30 days and were partially (10 animals per sex and dose group) observed for another 14 days without treatment.


Cage side observations, food consumption, body weight development, haematology, clinical biochemistry, urine analysis, macroscopic investigations, organ weights and histopathology of selected organs did not reveal any substance related toxic effects. The NOAEL in this study was 1000 mg/kg bw.


 


This OECD 422 study is a valid investigation of the toxicological effects resulting from repeated oral-gavage administration of the test item. The test item was administered in vehicle (1.0% CMC / 0.05% Tween 80 in highly purified water) at dosages of 100, 300, and 1000 mg/kg body weight/day, animals in control groups received the vehicle only. Test item was administered to male rats for 32 days and to female rats for 14 days prior to pairing, through the pairing and gestation periods until the F1 generation reached day 4 post partum.


Under the conditions of this study, no adverse effects were found in males or females up to the highest dose level of 1000 mg/kg bw/day.


 


A 90-day toxicity study in Fischer F344 rats was performed with the close analogue PY 74. Groups of 10 male and 10 female rats received 0, 50, 200 and 1000 mg/kg/day by oral gavage in corn oil for 90 days. The study included additional control and high dose groups of 6 males and 6 females each analyzed after a 4-week recovery period. All animals survived until scheduled termination without any toxic signs in life. In haematology, clinical biochemistry and urinalysis some scattered significant differences, altogether without a clear dose response and all only minor in severity, were noted. Post mortem examination including histopathology did not reveal any test substance related toxic alterations. The test substance caused elevated liver weights in the high dosed females. As there were no corresponding histopathological or clinical-biochemical alterations found, the most likely interpretation is an adaptive response by enzyme induction. No parallel trend was present in the males. The effects did not persist until the end of the recovery period. No other test substance-related effect was noted. The No-observed-adverse-effect-level (NOAEL) of the test item was 1000 mg/kg bw/day in both sexes.


 


Inhalation


Based on the results observed in a subchronic inhalation toxicity study according to OECD TG 413, it is concluded that the No Observed Adverse Effect Concentration (NOAEC) of test item was found to be 0.03 mg/L (30 mg/m³) when exposed for 6 hours/day, 5 days/week, for 13 weeks by flow-past nose-only inhalation route to Sprague Dawley rats. The nominal dose of 0.03 mg/L (6 hrs/day, 5 days/week) corresponded to an actual exposure concentration in males and females

Key value for chemical safety assessment

Toxic effect type:
dose-dependent

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Referenceopen allclose all

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 19 FEB 1979 to 03 APR 1979
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: basic data given, comparable to guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
Deviations:
not applicable
GLP compliance:
no
Remarks:
study was performed previous to GLP implementation
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Strain: WISKf (SPF 71)
- Source: in house breeding; Hoechst, breeding colony, Hattersheim
- Weight at study initiation: males: 73.8 +/- 2.29 g (69 g - 79 g); females: 73.6 +/- 2.41 g (69 g - 78 g)
- Fasting period before study: no
- Housing: 5 animals/sex/cage
- Diet: Standard diet Altromin 1324 (Altromin GmbH, Lage, Germany), ad libitum
- Water: tap water, ad libitum
- Acclimation period: no data


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-28
- Humidity (%): 25-50


Route of administration:
oral: gavage
Vehicle:
other: suspension in starch mucilage
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
- the test item suspension was freshly prepared every day before dosing
- 10% test item suspension in 2% starch mucilage, which was permanently stirred during the dosing procedure
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
30 days
Frequency of treatment:
daily except on weekends, i.e. 22 doses over a period of 30 days
Remarks:
Doses / Concentrations:
0, 1000 mg/kg bw
Basis:
actual ingested
No. of animals per sex per dose:
20
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: no lethality and no signs of toxicity were observed after single oral application of 5000 mg/kg bw
Positive control:
none
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: Yes
- Time schedule for examinations: one week before the first treatment, on the days of treatment, and two times per week (monday and thursday) during the recovery period

FOOD CONSUMPTION AND COMPOUND INTAKE (no feeding study): Yes
- Food consumption for each animal determined: Yes
- mean daily diet consumption calculated as % body weight: Yes

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: No data

WATER CONSUMPTION: Yes

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: weekly
- Dose groups that were examined: all

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at the end of the treatment period and at the end of the recovery period
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: 5 males and 5 females of each groupd
- Parameters examined: hematocrit, hemoglobin, erythrocytes count, total and differential leucocyte counts, median cell volume, median cell hemoglobin, platelet count, prothrombin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at the end of the treatment period and at the end of the recovery period
- Animals fasted: No data
- How many animals: 5 males and 5 females of each group
- Parameters examined: serum alkaline phosphatase (SAP), serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT)

URINALYSIS: Yes
- Time schedule for collection of urine: at the end of the treatment period
- Metabolism cages used for collection of urine: yes
- Animals fasted: yes
- Parameters examined: appearance, colour, protein, glucose, hemoglobin, bilirubin, pH-value, sediment


NEUROBEHAVIOURAL EXAMINATION: No
- weekly examinations for neurologic disturbances

Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes (liver, spleen, adrenals, kidney)
Other examinations:
ORGAN WEIGHTS:
- liver, spleen, adrenals, kidney
Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
no effects observed
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
SGPT: statistically significant decrease in females, fully reversible during the recovery period, no histopathological findings, therefore not regarded to be adverse
Urinalysis findings:
no effects observed
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not examined
Details on results:
CLINICAL SIGNS AND MORTALITY
- no effects

BODY WEIGHT AND WEIGHT GAIN
- no differences between treated animals and controls

FOOD CONSUMPTION
-no effects

WATER CONSUMPTION
- no effects

OPHTHALMOSCOPIC EXAMINATION
- not examined

CLINICAL CHEMISTRY
- SGPT statistically significantly decreased in females at the end of the treatment period, effects were fully reversible at the end of the recovery period; these effects were not regarded to be adverse as no histopathological findings were observed
- no effects in males or on other endpoints in females

HAEMATOLOGY
- no pathological findings

URINALYSIS
- no effects
- no discolouration of the urine observable

NEUROBEHAVIOUR
- no neurologic disturbances

ORGAN WEIGHTS
- no effects

GROSS PATHOLOGY
- no effects

HISTOPATHOLOGY: NON-NEOPLASTIC
- no effects

Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other:
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Critical effects observed:
not specified

The test item was excreted via faeces but not via urine (no discolouration of the urine).

Conclusions:
The test item did not reveal any substance related toxic effects under the conditions tested. The NOAEL in this subacute oral study was 1000 mg/kg bw in male and female rats.
Executive summary:
Male and female Wistar rats (20 animals per sex and dose group) received 22 daily oral (gavage) doses of 0 or 1000 mg test item per kg bw over a period of 30 days and were partially (10 animals per sex and dose group) observed for another 14 days without treatment.

Cage side observations, food consumption, body weight development, haematology, clinical biochemistry, urine analysis, macroscopic investigations, organ weights and histopathology of selected organs did not reveal any substance related toxic effects. The NOAEL in this study was 1000 mg/kg bw.

Endpoint:
short-term repeated dose toxicity: oral
Remarks:
combined repeated dose and reproduction / developmental screening
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 09 FEB 2012 and 03 JUL 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study (OECD 422) and in compliance with GLP.
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS

- Animals: Rat, RccHanTM: WIST(SPF)
- Rationale: Recognized by international guidelines as a recommended test system.
- Breeder: Harlan Laboratories, Inc., Maasheseweg 87c, 5800 AN Vernay / Netherlands
- Number of Animals: 44 males (11 per group) and 44 females (11 per group)
- Age (at Start of Treatment): 11 weeks
- Body Weight Range (at Start of Treatment): 315 to 369 g (males) and 187 to 243 g (females)
- Identification: parent animals had cage card and individual animal number (ear tattoo), pups were individually tattooed with Indian ink: on day 1 post partum.
- Randomization: Performed after at least three days of acclimatization using a computer-generated random algorithm. Body weights (recorded on the day of allocation) were taken into consideration in order to ensure similar mean body weights in all groups.
- Acclimatization: Under test conditions after health examination. Only animals without any visible signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS

- Conditions: Standard laboratory conditions. Air-conditioned with 10 - 15 air changes per hour, continuously monitored environmental conditions (temp. range: 22 ± 3 °C; relative humidity range: 30 - 70%). There was a 12 hour fluorescent light / 12-hour dark cycle with music during the light period.
- Accommodation: In groups of three to five animals in Makrolon type-4 cages with wire mesh tops up to the day of randomization and afterwards individually in Makrolon type-3 cages with wire mesh tops and sterilized standard softwood bedding (‘Lignocel’ J. Rettenmaier & Söhne GmbH & CoKG, 73494 Rosenberg / Germany, imported by Provimi Kliba SA, 4303 Kaiseraugst / Switzerland) with paper enrichment (ISO-BLOX from Harlan Laboratories B.V., Netherlands) batch/lot nos. 02105111001, 02105111201, 100099 and S211008A63972). During the pre-pairing period, cages with males were interspersed amongst those holding females to promote the development of regular estrus cycles.
- Diet: Pelleted standard Harlan Teklad 2018C (batch no. 80/11) rodent maintenance diet (Provimi Kliba SA, 4303 Kaiseraugst / Switzerland) was available ad libitum. Results of representative analyses for contaminants were included in the report as an Appendix.
- Water: Community tap-water from Füllinsdorf was available ad libitum in water bottles. Results of bacteriological assay, chemical and contaminant analyses of representative samples were included in the report as an Appendix.
Route of administration:
oral: gavage
Vehicle:
other: 1.0% CMC / 0.05% Tween 80 in highly purified water
Details on oral exposure:
DOSE FORMULATIONS

The dose formulations were prepared weekly using the test item as supplied by the Sponsor.

The test item was weighed into a glass beaker on a tared precision balance. Appropriate amount of the vehicle was weighted and added to the test item (w/w). Dose formulations were mixed on a magnetic stirrer for approximately 2 hours until mixtures were homogeneous. Separate formulations were prepared for each concentration.

Homogeneity of the test item in the vehicle was maintained during the daily administration period using a magnetic stirrer.

STORAGE OF DOSE FORMULATIONS

Dose formulations were stored at room temperature (20 ± 5 °C) in glass beakers.

Based upon the results of stability analyses performed within the non GLP Harlan Laboratories study 14-Day Oral Toxicity (Gavage) Study in the Wistar Rat, dose formulations are stable for at least 7 days if stored at room temperature.

TREATMENT

- Method: Oral, by gavage
- Rationale for Method: Administration by gavage is a common and accepted route of exposure for this type studies.
- Frequency of Administration: Once daily
- Target Dose Levels: 0 mg/kg bw/day (control group), 100 mg/kg bw/day (group 2), 300 mg/kg bw/day (group 3) and 1000 mg/kg bw/day (Group 4)
- Rationale for Dose Level Selection: The dose levels were selected based on a previous dose range-finding toxicity study in Han Wistar rats, Harlan Laboratories Study, using dose levels of 0, 100, 300 and 1000 mg/kg/day, where no adverse effects were observed up to the highest dose level.
- Dose Volume: 10 mL/kg body weight
- Dose Concentrations: 0 mg/mL (control group), 10 mg/mL (group 2), 30 mg/mL (group 3) and 100 mg/mL (group 4)
- Duration of Acclimatization Period: 7 days
- Duration of Treatment Period: 32 days males and approximately 7 weeks females
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
On the first treatment day samples from the control group as well as three samples (top, middle and bottom) of about 2 g of each concentration were taken prior to dosing for analysis of concentration and homogeneity. To confirm the stability (7 days) samples of about 2 g of each concentration were taken from the middle only of each aliquot used on day 7 of the treatment. During the sixth week of the treatment samples of about 0.5 g were taken to repeat the measurement of concentration, homogeneity and stability (7 days). During the last week of the treatment, samples were taken from the middle to confirm concentration. The aliquots for analysis of dose formulations were frozen (-20 ± 5 °C) and stored at -20 ± 5 °C until analysis.

The samples were analyzed by UV-VIS measurement following an analytical procedure developed by Harlan Laboratories. The test item was used as the analytical standard.

The following results were obtained:

Blank samples showed no absorbance and, therefore, it was confirmed that only 1.0% CMC / 0.05% Tween 80 in highly purified water applied within the control experiment.

The application formulations investigated during the study were found to comprise the tets item in the range of 83.0% to 111.3% and, thus, the required content limit of ±20% with reference to the nominal content was met. The homogeneous distribution of tets item in the preparations was approved because single results found did not deviate more than 9.8% (acceptance criterion: <15%) from the corresponding mean.

In addition, the test item was found to be stable in application formulations when kept seven days at room temperature due to recoveries which met the variation limit of 10% from the time-zero (homogeneity) mean, except for group 2 and 3 prepared on 16-Feb-2012 that exceeded the acceptance criteria. In application formulations of group 2 (10 mg/mL), the maximum deviation of time-zero mean was found to be 11.8%, and in application formulations of group 3 (30 mg/mL), the maximum deviation of time-zero mean was found to be 35.8%. However, there are some hints of what could be the cause. On the first and 7th treatment day as well as backup samples (date of preparation 16-Feb-2012), erroneously high amount of samples (about 2 g instead 0.5 g) were taken for analytical measurements making it difficult to work up. That was considered to probably be the reason for the differences as a second time using small amount of sample (0.5 g) shows the homogeneity and stability of the application formulation.

In conclusion, the results indicate the accurate use of the test item and 1.0% CMC / 0.05% Tween 80 in highly purified water as vehicle during this study. Application formulations were found to be homogeneously prepared and sufficient formulation stability under storage conditions was approved.
Duration of treatment / exposure:
MALES

32 days

FEMALES

about 7 weeks
Frequency of treatment:
once daily
Remarks:
Doses / Concentrations:
0 mg/kg bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
100 mg/kg bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
300 mg/kg bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
1000 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose:
11
Control animals:
yes, concurrent vehicle
Details on study design:
MALES

- Acclimatization: 7 days
- First Test Item Administration: Day 1 of pre-pairing
- Pre-Pairing: 14 days
- Pairing: 14 days maximum
- Treatment End: On day before sacrifice
- Blood Sampling: At Termination
- Necropsy: After 32 days of treatment, when no longer needed for assessment of reproductive effects

FEMALES

- Acclimatization: 7 days
- First Test Item Administration: Day 1 of pre-pairing
- Pre-Pairing: 14 days
- Pairing: 14 days maximum
- Gestation: Approximately 21 days
- Treatment End: On day 4 post partum
- Blood Sampling: On day 5 post partum
- Necropsy: On day 5 post partum (pups on day 4 post partum)
Positive control:
not required
Observations and examinations performed and frequency:
VIABILITY/MORTALITY

Twice daily

CLINICAL SIGNS

Daily cage-side clinical observations (once daily, during acclimatization and up to day of necropsy). Additionally females were observed for signs of difficult or prolonged parturition, and behavioral abnormalities in nesting and nursing.

FOOD CONSUMPTION

Males: on days 1 - 4, 4 - 8, 8 - 11 and 11 - 14 during pre-pairing period and weekly during after pairing period.
Females: on days 1 - 4, 4 - 8, 8 - 11 and 11 - 14 during pre-pairing period; on days 0 - 7, 7 14 and 14 - 21 during gestation period and on days 1 - 4 of during lactation period.

No food consumption was recorded during the pairing period.

BODY WEIGHTS

Recorded daily from treatment start to day of necropsy.

DETAILED CLINICAL OBSERVATIONS

Detailed clinical observations were performed outside the home cage in all animals. In males, it was performed once prior to the first administration of the test item and weekly thereafter. In females, it was performed once prior to the first administration of the test item, weekly during the pre-pairing and pairing periods and on days 0, 6, 13 and 20 of the gestation period.

Animals were observed for the following: changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions, and autonomic activity (e.g. lacrimation, piloerection, pupil size, unusual respiratory pattern). Changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies or bizarre behavior were also reported.

FUNCTIONAL OBSERVATIONAL BATTERY

At one time during the study (males shortly before the scheduled sacrifice and females on day 4 post partum) relevant parameters were performed with 5 P generation males and 5 P generation females from each group. This FOB assessment was conducted following the daily dose administration. Animals were observed for the following:
- Cage-side observations: faeces-balls, urine and posture as well as resistance to removal.
- Hand-held observations: muscle tone, constituation, skin, pupile size, palpebral closure, lacrimation, salivation, reaction to handling and general abnormalities.
- Open field observations: level of ambulatory activity including rearing (one minute evaluation), unusual body movements (e.g. spasms, convulsions), gait evaluation, behavior, hair coat, respiration, quantity of faeces-balls and urine.
- Reflexes: blinking, palpebral closure, pinna reflex, extensor thrust response, paw pinch, responsiveness to sharp noise, righting reflex and hearing ability (Preyer’s reflex).
- Measurements / Counts: hind limb / fore limb grip strength, landing foot splay, rectal temperature.

Any abnormal findings were recorded and, where appropriate, graded in severity.

Additionally, locomotor activity was measured quantitatively for the same animals. Activity was measured with an Activity Monitor AMS-0151 (FMI, Germany). Activity of the animals (based on beam count) was recorded for 6-minute intervals over a period of 30 minutes. These data and the total activity over 30 minutes were reported.

CLINICAL LABORATORY INVESTIGATIONS

Blood samples were obtained on the day of the scheduled necropsy from 5 males of each group. Blood samples from 5 lactating females of each group were obtained at the end of the pre-pairing period. Blood samples were drawn from the retro-orbital plexus of all animals under light isoflurane anesthesia. The animals were fasted for approximately 18 hours before blood sampling but allowed access to water ad libitum. The samples were collected early in the working day to reduce biological variation caused by circadian rhythms.

The following hematology parameters were determined:
- Erythrocyte count
- Hemoglobin
- Hematocrit
- Mean corpuscular volume
- Red cell volume distribution width
- Mean corpuscular hemoglobin
- Mean corpuscular hemoglobin concentration
- Hemoglobin concentration distribution width
- Leukocyte count, total
- Differential leukocyte count
- Platelet count
- Prothrombin time (= Thromboplastin time)
- Activated partial Thromboplastin time

The following clinical biochemistry parameters were determined:
- Glucose
- Urea
- Creatinine
- Bilirubin, total
- Cholesterol, total
- Triglycerides
- Aspartate aminotransferase
- Alanine aminotransferase
- Alkaline phosphatase
- Gamma-glutamyl-transferase
- Bile acids
- Sodium
- Potassium
- Chloride
- Calcium
- Phosphorus
- Protein, total
- Albumin
- Globulin
- Albumin/Globulin ratio

URINALYSIS

The following urinalysis parameters were determined in five males of each group, which are allocated to the blood analysis, during the last week of the study using timed urine volume collection:
- Volume (18 hours)
- Specific gravity (relative density)
- Color
- Appearance
- pH
- Nitrite
- Osmolality
- Protein
- Glucose
- Ketones
- Urobilinogen
- Bilirubin
- Blood/Blood cells
Sacrifice and pathology:
TERMINATION AND NECROPSY

Males were sacrificed after treatment for 32 days, when no longer needed for the assessment of reproductive effects. Dams were sacrificed on day 5 post partum. If birth did not occur on the expected date (day 21 post coitum), the dam was sacrificed on day 25 post coitum.

All animals sacrificed were subjected to a detailed macroscopic examination. Specimens of abnormal tissue were fixed in neutral phosphate buffered 4% formaldehyde solution. At the scheduled sacrifice, all animals were sacrificed by an injection of sodium pentobarbital. All P generation animals were exsanguinated. All parent animals were examined macroscopically for any structural changes. For the parent animals, special attention was directed at the organs of the reproductive system.The number of implantation sites and corpora lutea was recorded for all dams with litters. The uteri of non-pregnant females were placed in a solution of ammonium sulfide to visualize possible hemorrhagic areas of implantation sites.

SEMINOLOGY AND SPERMATID COUNT

Sperm analysis was performed on the first 5 males per group.

Motility:
At necropsy of adult males an epididymal sperm sample was obtained from the left cauda epididymidis of each male. The sample was diluted with a pre-warmed (about 35 °C) physiological medium, and shortly after being obtained, one hundred sperm was counted microscopically for determination of percentage of not motile, stationary motile and progressively motile sperm.

Morphology:
A second sperm sample from the left cauda epididymidis was used for morphological assessment after fixation and Eosin staining. 500 sperm per sample was evaluated microscopically and classified into the following categories:
A: Normal, complete sperm
B: Normal head only (tail detached)
C: Complete sperm, misshapen hook
D: Complete sperm, abnormally curved hook
E: Complete sperm, reversed head
F: Abnormal head only (tail detached)

Morphological sperm evaluation was performed only for group 1 and 4 males. In the absence of a treatment-related effect the slides for the group 2 and 3 males were not evaluated.

Sperm, Spermatid Count:
The left caudal epididymis and left testis was taken for determination of homogenization-resistant spermatids and caudal epididymal sperm reserve. These tissues were frozen at -20 ± 5°C pending evaluation. For evaluation the weighed tissues were placed in Triton-X-100 solution and homogenized with a blender (Ultra Turrax) and an ultrasonic water bath. Sperm or spermatid heads were counted microscopically using a modified Neubauer chamber. These evaluations were performed in the first instance only for group 1 and 4 males. In the absence of a treatment-related effect the remaining frozen tissues were not evaluated.

ORGAN WEIGHTS

At the scheduled sacrifice, testes and epididymides from all parental males were weighed separately. In addition, from 5 males and 5 females sacrificed at the end of the study which were selected from each group, the following organs were trimmed from any adherent tissue, as appropriate, and their wet weight taken.
- Adrenal glands (weighed as pairs)
- Brain
- Heart
- Kidneys (weighed as pairs)
- Liver
- Thymus
- Spleen

TISSUE PRESERVATION

The following tissues from all parental males were preserved in neutral phosphate buffered 4% formaldehyde solution:
- Prostate
- Seminal vesicles with coagulating gland
- Testes (in Bouin’s fixative)
- Epididymides (in Bouin’s fixative)

The following tissues from all parental females were preserved in neutral phosphate buffered 4% formaldehyde solution:
- Ovaries

In addition, from 5 males and 5 females per group selected for organ weights, the following tissues were preserved in neutral phosphate buffered 4% formaldehyde solution:
- Gross lesions
- Brain (representative regions including cerebrum, cerebellum and pons)
- Spinal chord
- Small and large intestines (incl. Peyer’s patches)
- Stomach
- Liver
- Kidneys
- Adrenals
- Spleen
- Heart
- Thymus
- Thyroids, and parathyroids if possible
- Trachea and lungs (preserved by inflation with fixative and then immersion)
- Uterus (with vagina)
- Urinary bladder
- Lymph nodes (mesenterial, mandibular)
- Peripheral nerve (sciatic)
- Bone marrow

HISTOTECHNIQUE

All organ and tissue samples to be examined by the principal investigator for histopathology phase were processed, embedded and cut at an approximate thickness of 2 - 4 micrometers and stained with hematoxylin and eosin. Additionally, the testis was stained by PAS-hematoxylin.

HISTOPATHOLOGY

Slides of all organs and tissues listed collected at terminal sacrifice from the animals of the control and high-dose groups were examined by the principal investigator. The same applied to all occurring gross lesions. Special emphasis was made on the stages of spermatogenesis and histopathology of interstitial cell structure.

Histological examination of ovaries was carried out on any females that did not give birth. In addition, microscopic examination of the reproductive organs of all infertile males was made.

A histopathology peer review was performed. A histopathology phase report was provided by the principal investigator for inclusion in the main report as an appendix.
Other examinations:
MATING, GESTATION, LACTATION

During the pairing period, females were housed with sexually mature males (1:1) until evidence of copulation was observed. The females were removed and housed individually if: the daily vaginal smear was sperm positive or a copulation plug was observed. The day on which positive mating was determined (copulation plug or sperm) was designated day 0 post coitum. All dams were allowed to give birth and rear their litters (F1 pups) up to day 4 post partum. Day 0 was designated as the day on which a female had delivered all her pups.

REPRODUCTIVE AND OFFSPRING VIABILITY INDICES

From the on-line recorded reproduction data, the following parameters were calculated: fertility indices, mean precoital time, post-implantation losses, mean litter size, pup sex ratios and viability indices.

LITTER OBSERVATIONS

The litters were examined for litter size, live births, still births and any gross anomalies. The sex ratio of the pups was recorded. Pups were weighed individually (without identification) on days 0 (if possible), 1 and 4 post partum.

POSTMORTEM EXAMINATION OF OFFSPRING

Pups were sacrificed on day 4 post partum. All animals were sacrificedby by an injection of sodium pentobarbital and subjected to a detailed macroscopic examination. Specimens of abnormal tissue were fixed in neutral phosphate buffered 4% formaldehyde solution. Pups dead during the study, except those excessively cannibalized, were examined macroscopically.
Statistics:
The following statistical methods were used to analyze food consumption, body weights, reproduction data, grip strength, landing foot splay, body temperature, locomotor activity, hematology and biochemistry parameters, urinalysis, sperm analyses and organ weights:
- Means and standard deviations of various data were calculated and included in the report.
- The Dunnett-test (many to one t-test) based on a pooled variance estimate was applied if the variables can be assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
- The Steel-test (many-one rank test) was applied instead of the Dunnett-test when the data cannot be assumed to follow a normal distribution.
- Fisher's exact-test was applied if the variables can be dichotomized without loss of information.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Yellow stained faeces was noted in all males and females in dose groups; this was due to the staining properties of the test item.
Mortality:
mortality observed, treatment-related
Description (incidence):
Yellow stained faeces was noted in all males and females in dose groups; this was due to the staining properties of the test item.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Transient reduction in body weight gain was noted in males at the dose levels of 1000 and 300 mg/kg bw/day .
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
Locomotor activity was not affected and functional observational battery gave no indication of a test item-related effect.
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not examined
Details on results:
1. IN-LIVE DATA OF PARENTAL ANIMALS

VIABILITY / MORTALITY

All animals survived scheduled study period.

DAILY CLINICAL SIGNS OR OBSERVATIONS

Feces stained yellow were noted in all males and females in all dose groups starting from day 2 or 3 of the treatment until completion of the study. This observation was due to staining properties of the yellow coloured test item.

No further test item-related clinical signs or observations were noted in males or females at any dose level.

Incidentally, scabs on the neck were noted in one male in the control group.

FINDINGS AT DETAILED WEEKLY CLINICAL OBSERVATIONS

No test item-related findings were noted during weekly detailed clinical observations.

Incidentally, in one female at the dose level of 1000 mg/kg bw/day missing upper incisors on day 6 and 13 of the gestation period were recorded.

No further findings were noted in males or females in any dose group.

FUNCTIONAL OBSERVATIONAL BATTERY

None of the parameters under investigation during the functional observational battery gave an indication of a test item-related effect.

All findings recorded during the tests were considered not to be test item-related. Decreased or increased rearings, increased faeces balls and vocalization were noted in individual animals without dose dependency. At the dose level of 1000 mg/kg bw/day, one female was noted with multiple findings: reduced activity, ruffled fur, decreased rearings and salivation. Because of isolated occurrence, this observation was considered not to be related to the treatment.

Remaining parameters under investigation were similar in all groups.

LOCOMOTOR ACTIVITY

No effects on locomotor activity were noted in males or females at any dose level.

Mean beam counts during the 30 minutes of measurement at the dose levels of 0, 100, 300 and 1000 mg/kg bw/day were respectively: 1079, 1198, 1252 and 1207 in males and 965, 917, 853 and 867 in females.

FOOD CONSUMPTION OF MALES

No effects on food consumption were noted in males at any dose level.

Mean differences in food consumption at the dose levels 100, 300 and 1000 mg/kg bw/day were respectively: ±0.0%, -1.2% and -4.6% during the pre-pairing period and -2.0%, -2.0% and -0.8% during the after pairing period (percentages refer to the respective values in the control group).

FOOD CONSUMPTION OF FEMALES

No effects on food consumption were noted in females at any dose level.

Incidentally, statistically significantly higher food consumption was noted at the dose level of 100 mg/kg bw/day on days 8 - 11 of the pre-pairing period and at the dose level of 300 mg/kg bw/day on days 7 - 14 of the gestation period. In the absence of any dose dependency, these differences were not related to the treatment.

Mean differences in food consumption at the dose levels 100, 300 and 1000 mg/kg bw/day were respectively: +8.0%, ±0.0%, and +3.4% during the pre-pairing period, +1.8%, +6.3% and +3.1% during the gestation period and +11.7%, -3.0% and +3.8% during the lactation period (percentages refer to the respective values in the control group).

BODY WEIGHTS OF MALES

At the dose levels of 1000 and 300 mg/kg bw/day, reduction in body weight gain was noted during the pre-pairing period; mean body weight gain within this period was +10% and +12% at the high- and mid-dose level respectively, compared to +14% in the control group. Reduction in body weight gain was statistically significant at the high-dose level starting from day 3 until the end of pre-pairing period and at the mid-dose level on days 7, 9, 11, 12 and 14 of the pre-pairing period. This effect was considered to be test item-related. During the pairing and after pairing periods, body weight gain was similar at all dose levels.
Body weights of males in all dose groups were similar to the respective control values during the entire study period.
Because the reduction in body weight gain at the high- and mid-dose levels was reversible and did not result in any significant differences in body weights, this observation was considered not to be adverse.

Mean differences in body weight gain at the dose levels of 0, 100, 300 and 1000 mg/kg bw/day were respectively: +14%, +13%, +12% and +10% during the pre-pairing period, +3%, +2%, +3% and 3% during the pairing period and +1%, +1%, +1% and +2% during the after pairing period (percentages refer to the body weight change within the respective period).

BODY WEIGHTS OF FEMALES

Body weights and body weight gain of females were not affected by the treatment with the test item at any dose level.

Mean differences in body weight gain at the dose levels of 0, 100, 300 and 1000 mg/kg bw/day were respectively: 8%, 9%, 8% and 8% during the pre-pairing period, 53%, 53%, 59% and 57% during the gestation period and 6%, 7%, 6% and 6% during the lactation period (percentages refer to the body weight change within the respective period).

2. CLINICAL LABORATORY INVESTIGATIONS

HEMATOLOGY

No test item-related effects on hematology parameters were noted in males or females at any dose level.

At the dose level of 300 mg/kg bw/day in males, amount of large unstained cells (LUC) was statistically significantly lower when compared to the control value. No reduction of this value occurred at the dose level of 1000 mg/kg bw/day and therefore the difference at mid-dose level was not related to the treatment.

CLINICAL BIOCHEMISTRY

No test item-related effects on biochemistry parameters were noted in males or females at any dose level.

At the dose level of 300 mg/kg bw/day in males, potassium concentration was statistically significantly higher when compared to the control value. No increase of this value occurred at the dose level of 1000 mg/kg bw/day and therefore the difference at mid-dose level was not related to the treatment.

In females following parameters were statistically significantly lower if compared to the respective control values: concentration of albumin at the dose level of 100 mg/kg bw/day, concentration of albumin and albumin to globulin ratio at the dose level of 300 mg/kg bw/day and concentration of total protein and albumin at the dose level of 1000 mg/kg bw/day. Changes of these values were not dose depended and all values were within historical control range. Therefore these changes were considered not to be test item-related.

URINALYSIS

No changes in urine parameters were noted in males at any dose level.

3. TERMINAL FINDINGS - PARENTAL ANIMALS

SEMINOLOGY AND SPERMATID COUNT

At the dose levels of 1000 and 300 mg/kg bw/day, statistically significant changes in motility of sperms were noted. Mean count of progressive sperms was reduced at the high dose level and it was 60.4% compared to 75.6% in the control group. This value is within the range of a limited pool of historical controls for OECD 422 (n=4) extended by OECD 416 data (n=5). Mean count of not motile sperms was increased and was 35.5% and 34.2% at the high- and mid-dose dose levels, respectively, compared to 20.2% in the control group. These changes might be test item-related. However no significant dose dependent trend indicated by probability values of <0.05 was determined for any of these changes when performing a linear regression analysis (least squares). In the absence of any findings during necropsy or histopathological examination of male reproductive organs as well as absence of any effects on reproduction, the differences in sperm motility were considered not to be adverse.

No further changes were noted during sperm analysis. At the high-dose level, all morphological categories of sperms were represented with similar frequency to that in the control group whereas sperm count was similar (testis) or slightly higher (epididymidis) when compared to the respective control values.

ORGAN WEIGHTS

No test item-related changes in organ weights were noted in males or females at any dose level.

At the dose level of 100 mg/kg bw/day in males, statistically significantly lower weight of testis was noted. This effect was due to reduced testis weights of one male (no. 15, malformation of testis in this male correlated with histopathological findings). No significant changes of testis weights were noted at the mid- and high dose levels and therefore these changes were not related to the treatment with the test item.

No further significant differences in absolute or relative organ weights were noted in males or females at any dose level.

MACROSCOPICAL FINDINGS

Type and distribution of findings noted during macroscopical examination of males and females did not indicate any test item related effect.

HISTOPATHOLOGY FINDINGS

Under the conditions of this experiment, the test item did not induce histopathological lesions. All findings recorded were within the range of normal background lesions which may be recorded in animals of this strain and age.
There were no abnormal lesions encountered during sperm staging regarding completeness of stages and maturation of cell populations. Individual lesions recorded were within the range of background alterations that may be recorded in this type of study, in rats of this strain and age.

During examination of reproduction organs of infertile males and females, bilateral total tubular degeneration of testes was found in male no. 15 at the dose level of 100 mg/kg bw/day. This lesion was considered to be the reason for infertility of male no.15. No further findings correlated to the infertility were noted in reproduction organs of the remaining infertile males or females.
Key result
Dose descriptor:
NOAEL
Remarks:
for general toxicity (P)
Effect level:
1 000 mg/kg bw/day (actual dose received)
Sex:
male
Basis for effect level:
other: Transiently lower body weight gain noted at the dose levels of 1000 and 300 mg/kg bw/day was considered not to be adverse.
Key result
Dose descriptor:
NOAEL
Remarks:
for general toxicity (P)
Effect level:
1 000 mg/kg bw/day (actual dose received)
Sex:
female
Basis for effect level:
other: No toxicologically relevant findings were noted in females up to the dose level of 1000 mg/kg bw/day, the highest dose level used.
Key result
Dose descriptor:
NOAEL
Remarks:
for reproduction (P)
Effect level:
1 000 mg/kg bw/day (actual dose received)
Sex:
male/female
Basis for effect level:
other: Reduced motility of sperms noted at the dose levels of 1000 and 300 mg/kg bw/day was considered not to be adverse.
Key result
Dose descriptor:
NOAEL
Remarks:
for development (F1)
Effect level:
1 000 mg/kg bw/day (actual dose received)
Sex:
male/female
Basis for effect level:
other: Under the condition of the study, post-implantation loss, body weights of pups and results of examination of pups did not indicate developmental effects up to the dose level of 1000 mg/kg bw/day, the highest dose level used.
Key result
Critical effects observed:
not specified
1. REPRODUCTION, BREEDING AND PUP DATA

 

SUMMARY OF PERFORMANCE 

P Animals Breeding for F1 Litters 

 

Group
(mg/kg/day)

1
(0)

2
(100)

3
(300)

4
(1000)

Female numbers

45-55

56-66

67-77

78-88

Number of females paired

11

11

11

11

Number of females mated

11

11

11

11

Number of non pregnant females (A)

2

1

1

2

Numbers of pregnant females, which did not deliver any pups (B)

0

0

0

1

Number of females which reared their pups until day 4 post partum

9

10

10

8

(A) Female nos. 46,50, 59, 69, 83 and 85

(B) Female no. 87

 

MATING PERFORMANCE AND FERTILITY

 

Mating performance and fertility were not affected by the treatment at any dose level.

  

Percentage of mating was 100% in all groups. Mating of all females was recorded during the first seven days of the pairing period.

 

Mean (median) precoital times were 2.9 (3), 2.6 (3), 2.9 (3) and 2.8 (3) days at the dose levels of 0, 100, 300 and 1000 mg/kg bw/day, respectively.

 

Six females were not pregnant: two in the control group, one at each dose level 100 and 300 mg/kg bw/day and two at the dose level of 1000 mg/kg bw/day. Consequently, fertility indexes (number of females pregnant as percentages of females paired) and conception rate (number of females pregnant as percentages of females mated) were 81.8%, 90.9%, 90.9% and 81.8% at the dose levels of 0, 100, 300 and 1000 mg/kg bw/day, respectively.

 

One female at the high dose level had two implantation sites but delivered no pups. Consequently, gestation index (number of females with living pups as percentages of females pregnant) was 88.9% at the high-dose level and 100% in remaining groups.

 

CORPORA LUTEA COUNT

  

No test item-related effects on corpora lutea count were observed at any dose level.

 

Mean number of corpora lutea per dam was 13.6, 15.5, 17.4 and 14.4 in order of ascending dose levels.

 

At the dose level of 300 mg/kg bw/day, mean number of corpora lutea per dam was statistically significantly higher when compared to the control group. In the absence of any dose dependency, this observation was considered not to be related to the treatment.

 

DURATION OF GESTATION

  

No effects on duration of gestation were observed at any dose level.

 

Mean duration of gestation was 21.6, 21.8, 21.8 and 21.6 days, in order of ascending dose level.

 

IMPLANTATION RATE AND POST-IMPLANTATION LOSS

 

No effects on implantation rate and post-implantation loss were observed at any dose level.

 

In order of ascending dose levels, mean number of implantations was 13.0, 13.0, 14.2 and 14.4 per dam whereas mean incidence of post-implantation loss was 1.0, 1.0, 1.4 and 0.4 per dam.

 

LITTER SIZE AT FIRST LITTER CHECK

 

No effects on litter size were noted at any dose level.

During the first litter check, one dead pup was found in a litter at the dose level of 1000 mg/kg bw/day. Because of isolated occurrence, this finding was considered to be incidental.

 

Mean number of living pups per dam at first litter check was 12.0, 12.2, 12.8 and 14.1 in order of ascending dose levels.

Birth index (number of pups born alive as a percentage of implantations) was 92.3%, 92.2%, 90.1% and 97.0% at the dose level of 0, 100, 300 and 1000 mg/kg bw/day.

 

POSTNATAL LOSS DAYS 0 - 4 POST PARTUM

 

No test item-related effects on postnatal loss were noted at any dose level.

 

In the control group one pup was missing on day 4, at the low-dose level one pup was missing on day 2, at the mid dose level eight pups (from 2 litters) were missing on day 4 and at the high dose level two pups (from two litters) were missing on day 2 of the lactation period. Mean postnatal loss during four days of lactation was 0.1, 0.1, 0.8 and 0.3% at the dose level of 0, 100, 300 and 1000 mg/kg bw/day, respectively. Consequently, viability index (number of pups alive at termination on day 4 p.p. as a percentage of pups born alive) was 99.1, 99.2, 93.8 and 98.2 in order of ascending dose levels.

 

At the dose level of 300 mg/kg bw/day, total number of pups lost was statistically significantly higher and viability index was statistically significantly lower when compared to the respective control values. These observations were due to a loss of seven pups in one litter. Because higher mortality of pups was noted only in one litter and in the absence of increased mortality of pups at the dose level of 1000 mg/kg bw/day, this observation was considered not to be related to the treatment.

 

EXTERNAL EXAMINATION AT FIRST LITTER CHECK AND DURING LACTATION

 

No test item-related findings were noted in pups during first litter check and during lactation at any dose level.

 

In the control group, missing tail tip was noted at first litter check and during lactation in one pup. At the dose level of 300 mg/kg bw/day, all pups in one litter had reduced temperature on day 3 and several pups from this litter had reduced temperature on day 4 of the lactation period. At the dose level of 1000 mg/kg bw/day, one pup, which was dead at first litter check had no milk in the stomach and was partially cannibalized.

 

No further findings were noted in pups at any dose level.

 

SEX RATIOS

 

Pups sex ratio was not affected by exposure to the test item at any dose level.

At first litter check, percentages of male pups were 48%, 48%, 38% and 49% at the dose levels of 0, 100, 300 and 1000 mg/kg bw/day.

 

BODY WEIGHTS TO DAY 4 POST PARTUM

 

Body Weights and body weight gain of pups were not affected by the treatment with the test item at any dose level.

 

Mean body weights of pups on day 1 post partum were: 8.9 g, 9.2 g, 8.9 and 8.2 g and mean differences in body weights during lactation were +47.4%, +47.7%, 43.6% and +38.7%, at the dose levels of 0, 100, 300 and 1000 mg/kg/day, respectively.

 

At the dose level of 1000 mg/kg bw/day, slightly not statistically significantly lower body weight gain of pups was noted. This effect was considered to be due to a higher number of pups at this dose level which was supported by observation that reduction of body weight gain was more pronounced in litters of higher size. Therefore this effect was not test item-related.

 

MACROSCOPICAL FINDINGS

 

No findings were found during necropsy of pups in any dose group.

 

Conclusions:
This study is a valid investigation of the toxicological effects resulting from repeated oral-gavage administration of the test item. The test item was administered in vehicle (1.0% CMC / 0.05% Tween 80 in highly purified water) at dosages of 100, 300, and 1000 mg/kg body weight/day, animals in control groups received the vehicle only. Test item was administered to male rats for 32 days and to female rats for 14 days prior to pairing, through the pairing and gestation periods until the F1 generation reached day 4 post partum.

Under the conditions of this study, no adverse effects were found in males or females up to the highest dose level of 1000 mg/kg bw/day.

All animals survived the scheduled study period.
During the treatment, faeces stained yellow were noted in all males and females receiving test item. This observation was due to staining properties of the test item.

At the dose levels of 1000 and 300 mg/kg bw/day in males, statistically significantly reduced body weight gain was noted during the pre-pairing period. During the remaining study period body weight gain at the high- and mid-dose levels was similar to the control values. Body weights at these dose levels were similar to the control values during the entire study period. Because reduction of body weight gain was reversible and no statistically significant differences in body weights were noted at the high- and mid-dose levels, this effect was considered not to be adverse.

No further test item related observations were noted in males or females at any dose level during the live part of the study.

Terminal examinations revealed changes in motility of sperms at the dose levels of 1000 and 300 mg/kg bw/day. Statistically significant reduction in mean count of progressive sperms and increase in mean count of not motile sperms were noted at the high dose level and statistically significant increase in mean count of not motile sperms was noted at the mid-dose level. No further effects on male reproductive system were noted during the study. Sperm morphology and sperm count at the high-dose level was similar to the control values. Weights of male reproductive organs, macroscopical and histopathological examination of testes and epididymides gave no indication of any treatment-related effect. Further, no indication of effects on reproduction was noted within this study up to and including the highest dose level. For this reason, changes in motility of sperms were considered not to be adverse.

Reproduction and development were not affected by the treatment. Mating performance, fertility, duration of gestation, corpora lutea count, implantation rate, post implantation and postnatal loss or litter size were similar in the control and all dose groups. There were no test item-related findings in pups noted during the first litter check, the first 4 days post partum or during the necropsy, pups body weights and body weight gain were not affected by the treatment at any dose level.

Based on these results, the NOAEL (No Observed Adverse Effect Level) for general toxicity in males and females and for reproduction/developmental toxicity was considered to be 1000 mg/kg bw/day, the highest dose level used.
Executive summary:

The purpose of this study was to generate preliminary information concerning the effects of the test item on the possible health hazards likely to arise from repeated exposure over a relatively limited period of time. In addition it provides information on possible effects on male and female reproductive performance such as gonadal function, mating behavior, conception, development of the conceptus and parturition.

 

The tets item was administered to male rats for 32 days and to female rats for 14 days prior to pairing, through the pairing and gestation periods until the F1 generation reached day 4 post partum.

 

The following dose levels were applied:

 

Group 1:                         0 mg/kg body weight/day (control group)

Group 2:                     100 mg/kg body weight/day

Group 3:                     300 mg/kg body weight/day

Group 4:                    1000 mg/kg body weight/day

 

A standard dose volume of 10 mL/kg body weight with a daily adjustment to the actual body weight was used. Control animals were dosed with the vehicle alone (1.0% CMC / 0.05% Tween 80 in highly purified water).

 

The following results were obtained:

 

MORTALITY AND GENERAL TOLERABILITY OF PARENTAL ANIMALS

 

All animals survived the scheduled study period.

 

During the treatment, yellow stained faeces was noted in all males and females in dose groups. This observation was due to staining properties of the test item.

 

No further test item-related observations were noted during clinical daily or detailed weekly observations in males or females at any dose level.

 

FUNCTIONAL OBSERVATIONAL BATTERY AND LOCOMOTOR ACTIVITY OF PARENTAL ANIMALS

 

None of the parameters under investigation during the functional observational battery gave an indication of a test item-related effect.

 

Locomotor activity was not affected by the treatment with test item in males or females at any dose level.

 

FOOD CONSUMPTION OF PARENTAL ANIMALS

 

No effects on food consumption were noted in males or females at any dose level.

 

BODY WEIGHT OF PARENTAL ANIMALS

 

In males at the dose levels of 1000 and 300 mg/kg bw/day, dose dependent and statistically significant reduction in body weight gain was noted during the pre-pairing period. No differences in body weight gain were noted at any dose level during the remaining study period. Body weights of males in all dose groups were similar to the respective control values during the entire study period. Because reduction in body weight gain at the high- and mid-dose levels was reversible and did not cause significant changes in body weights, this effect was considered not to be adverse.

 

Body weights and body weight gain of females were not affected by the treatment at any dose level.

 

CLINICAL LABORATORY INVESTIGATIONS IN PARENTAL ANIMALS

 

No test item-related effects on hematology and clinical biochemistry parameters were noted in males or females at any dose level.

 

No changes in urine parameters were noted in males at any dose level.

 

REPRODUCTION AND BREEDING DATA

 

Mating performance, fertility, corpora lutea count, duration of gestation, implantation rate and post-implantation loss, litter size or postnatal loss were not affected by the treatment with the test item.

 

SEMINOLOGY AND SPERMATID COUNT IN PARENTAL ANIMALS

 

At the dose level of 1000 mg/kg bw/day, reduction in mean count of progressive sperms and increase in mean count of not motile sperms were noted. At the dose level of 300 mg/kg bw/day, increase in mean count of not motile sperms was noted. A significant dose dependent trend couldn't be established.

In the absence of any findings during necropsy or histopathological examination of male reproductive organs as well as absence of any effects on reproduction, the differences in sperm motility were considered not to be adverse.

 

Sperm morphology and sperm count were not affected by the treatment at any dose level.

 

ORGAN WEIGHTS OF PARENTAL ANIMALS

 

No test item-related changes in organ weights were noted in males or females at any dose level.

 

MACROSCOPICAL FINDINGS AND HISTOPATHOLOGICAL EXAMINATIONS OF PARENTAL ANIMALS

 

Type and distribution of findings noted during macroscopical examination did not indicate any test item related effect.

All findings recorded during histopathological examination were within the range of normal background lesions which may be recorded in animals of this strain and age.

 

FINDINGS IN PUPS AT FIRST LITTER CHECK AND DURING LACTATION

 

No test item-related findings were noted in pups during first litter check and during lactation at any dose level.

 

Pups sex ratio was not affected by exposure to the test item at any dose level.

 

PUP WEIGHTS TO DAY 4 POST PARTUM

 

Body Weights and body weight gain of pups were not affected by the treatment with the test item at any dose level.

 

MACROSCOPICAL FINDINGS IN PUPS

 

No findings were found during necropsy of pups in any dose group.

 

CONCLUSION

 

Based on these results, the NOAEL (No Observed Adverse Effect Level) for general toxicity in males and females and for reproduction/developmental toxicity was considered to be 1000 mg/kg bw/day, the highest dose level used.

 

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: performed in accordance with OECD and GLP guidelines
Reason / purpose for cross-reference:
reference to other study
Qualifier:
according to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.26 (Sub-Chronic Oral Toxicity Test: Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Deviations:
no
Principles of method if other than guideline:
Guideline study.
GLP compliance:
yes (incl. QA statement)
Limit test:
no
Species:
rat
Strain:
Fischer 344
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Strain: F344/DuCrl, SPF
- Source: Charles River Deutschland GmbH, 97633 Sulzfeld, Deutschland
- Age at first administration: ca. 7 weeks
- Weight at first administration: males: 165 g, females: 121 g
- Identification: By tattooing the individual animal number into the right pinna and cage tag.
- Housing: Optimal hygienic conditions. Makrolon cages Type IV (33 cm x 55 cm area, 20 cm height). The animals were caged in groups (3 or 4 animals of one group and one sex per cage). The first 4 animals of a group were housed together in one cage and the remaining ones in groups of 3 in two more cages.
- Diet: Ssniff R/M-H maintenance diet for rats and mice (item V1534-3 ) ad libitum.
- Water: Tap water, acidified with HCl to pH ¿3, from an automatic watering system, ad libitum.
- Nesting material: Nesting material and nibbling wood bricks (sized 10 cm x 2 cm x 2 cm), same material as the bedding material, were offered freshly once a week.
- Acclimation period: 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature: Average of 21.9 °C.
- Humidity: Average of 49 % .
- Air changes: 12 per hour.
- Photoperiod: Artificial light from 6 a.m. to 6 p.m.
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
The test substance was administered after being blended with (respectively dissolved in) the vehicle. Preparations of the test substance were made freshly every day shortly before the administration to the animals.

VEHICLE
- Justification for use and choice of vehicle (if other than water): The test substance was neither soluble in nor miscible with any of the commonly used aqueous vehicles (water, water with modified cellulose, with or without detergent). Therefore corn oil was used as vehicle, which allowed the preparation of suitable suspensions.
- Concentration in vehicle: Appropriate preparations were made to allow a uniform dose volume for all groups.
- Amount of vehicle: 5 mL per kg body weight.
- Purity: nutritional grade
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The test substance was analysed by a validated HPLC-method. A linear calibration line was established. The variance homogeneity was checked over the working range of 10.1 to 51.8 mg/L.
Stability: Determined on Day 1.
Test conditions: Ambient temperature, exposed to light. The concentrations of preparations for the low dosed group were checked. A loss of at most 5 % within 4 h was tolerated. The stability of higher concentrated suspensions is considered better than that of lower ones.
The test substance was found to be sufficiently stable in the preparations for at least 4 h.

Homogeneity: Determined on Days 1 and 37. 3 samples of 1.0 mL each of the preparations for the three dosed groups A, B and C+CS were taken using the syringe plus probe by the technician at the beginning, in the middle and at the end of the dosing procedure for the group concerned. A deviation of the individual samples from the mean of at most ± 10 % was tolerated.
Actual concentration: Determined on Days 1 and 37. The same samples as for the determination of homogeneity were used. A deviation of the mean of the 3 samples from the target concentration of at most ± 10 % was tolerated.
The concentrations and the homogeneity of the test substance in the preparations were within the chosen limits at both sampling terms.
Duration of treatment / exposure:
Day 1 was the day of the first administration of the test substance.
The animals of all groups were treated with the test substance solutions or with the vehicle once a day on 91 (males) or 92 (females) consecutive days.
Animals of the satellite groups KS and CS were kept after cessation of dosing without a further administration for 29 (females) or 30 (males) additional days.
Frequency of treatment:
Once a day.
Remarks:
Doses / Concentrations:
0 mg/kg: control group K and also control satellite group KS
Basis:
other: nominal, gavage
Remarks:
Doses / Concentrations:
50 mg/kg: low dose, group A
Basis:
other: nominal, gavage
Remarks:
Doses / Concentrations:
200 mg/kg: mid dose, group B
Basis:
other: nominal, gavage
Remarks:
Doses / Concentrations:
1000 mg/kg: high dose, group C and also high dose satellite group CS
Basis:
other: nominal, gavage
No. of animals per sex per dose:
10 males and 10 females in the main groups K, A, B and C.
6 males and 6 females in the satellite groups KS and CS.
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The doses chosen were derived from and based on the results of a dose range finding study. Groups of 5 male and 5 female rats each were administered 100 mg or 316 mg or 1000 mg test substance in 5 mL corn oil per kg body weight orally once a day for 14 consecutive days. Investigations performed: Animal observations, body weights and feed consumption, necropsy.
Summarized results: All animals survived until the terminal sacrifice. There were no treatment related observations in life. Body weights and feed consumption were not notably affected. All animals were found to be grossly normal at the post mortem examination.
1000 mg per kg body weight is the highest feasible dose for a subchronic toxicity study and was thus chosen as high dose. The low dose, where no toxic effects are anticipated and the mid dose were chosen by the sponsor.
- Rationale for animal assignment (if not random): randomisation procedure.
- Rationale for selecting satellite groups: A control group KS and a high dose group CS was used to investigate the development of changes after finishing of dosing.
- Post-exposure recovery period in satellite groups: 29 days for females and 30 days for males.
- Bias control: The sequence of animals for clinical observations, blood sampling, blood and urine analyses, necropsy and histopathology was randomised.
Positive control:
None.
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Days 0, 7, 14, 21, 28, 36, 42, 49, 56, 63, 70, 77, 85, 91: all animals of all groups. Days 98, 105, 112, 119: all animals of the satellite groups. Special emphasis was put on skin, fur, eyes, visible mucous membranes, incisors, secretion and excretion, body odour, autonomous activities (e.g. lacrimation, piloerection, pupillar size, abnormal breathing, and body surface temperature), vocalisation, abnormal locomotion, movements and posture, presence of convulsions or paralysis, stereotypes, bizarre behaviour, visible or palpable tissue masses.

BODY WEIGHT: Yes
- Time schedule for examinations: The individual body weights were determined:
Day 1 to 91: Once a week, all animals. Days 91 to 120: Satellite groups.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Determined per cage in weekly intervals in all animals.

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: No

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

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: Day 0: All animals of all groups. Day 86: All animals of all groups. Examination of the right eyes of all animals by adspection and by fundus examination, in case of the presence of lesions, and if considered as necessary, additionally by a slit lamp examination
An additional ophthalmoscopical examination at the end of the recovery period in the animals of groups KS and CS was omitted, as no test substance related alterations were noted at the end of the dosing period.
- Dose groups that were examined: each group.

HAEMATOLOGY: Yes
- Blood samples were taken from the retrobulbar vein plexus of the left eyes in the morning.
- Time schedule for collection of blood: Blood was taken on Day 92 from all male animals of groups K, A, B and C, on Day 93 from all female animals of groups K, A, B and C and of all animals of groups KS and CS on Day 121.
- Anaesthetic used for blood collection: Yes, slight ether anaesthesia
- Animals fasted: Yes. Feed was offered again immediately after the blood sampling.- Parameters determined (abbreviations, when commonly used):
¿ Red blood cell count (RBC)
¿ Haemoglobin concentration (HGB)
¿ Haematocrit (HCT)
¿ Mean corpuscular haemoglobin (MCH)
¿ Mean corpuscular haemoglobin concentration (MCHC)
¿ White blood cell count (WBC)
¿ Mean cell volume (MCV)
¿ Platelet count (PLT)
¿ Differential white blood cell count (% of the different cell species)
¿ Prothrombin time (Quick) as an indicator of blood clotting capacity.

CLINICAL CHEMISTRY: Yes
- Blood samples were taken from the same animals and at the same time as for haematology. Plasma (with Li-heparin as anticoagulant) was obtained by centrifugation of the blood samples after incubation at 37 °C for about 1 hour.
- Parameters determined (abbreviations, when commonly used):
¿ alanin aminotransferase (ALT, GPT)
¿ albumin (ALB)
¿ alkaline phosphatase (AP)
¿ aspartate aminotransferase (AST, GOT)
¿ cholesterol (CHOL)
¿ creatinine (CREA)
¿ gamma-glutamyltransferase (GGT)
¿ glucose (GLU)
¿ potassium (K)
¿ sodium (Na)
¿ total protein (TP)
¿ urea (UREA)

URINALYSIS: Yes
- Time schedule for collection of urine:
Urine was collected overnight by placing the animals in metabolism cages and cooling the urine with ice.
Day 91/92: All males of groups K, A, B and C.
Day 92/93: All females of groups K, A, B and C.
Day 120/121: All animals of groups KS and CS.
- Parameters determined:
¿ total amount (weighed),
¿ colour, turbidity, appearance (visual),
¿ osmolarity (Osmometer)
¿ pH (pH-Meter),
¿ proteins, glucose, ketones, urobilinogen, bilirubin, erythrocytes, haemoglobin (semi-quantitative, test sticks),
¿ sediment analysis (microscopically).
- Animals fasted: Yes. Animals were given continuous access to drinking water, but no feed was offered during the urine collection time.

NEUROBEHAVIOURAL EXAMINATION: Yes.
Assessment of the behaviour, the motor activities, and the sensory reactivity to different stimuli (acoustic, tactile, visual and proprioceptive) was performed outside the home cage in a standard arena.
On Day 87: all male animals of all groups.
On Day 91: all female animals of all groups.
On Day 119: all animals of groups KS and CS.
The eyelid and auricular reflexes (tactile and proprioceptive) were tested by slightly touching the cornea and the interior of a pinna with a nylon cord of approximately 0.6 mm diameter. Shaking of the head was considered as positive response.
Acoustic reactivity was tested by response to a moderate sound (clapping of the hands). Twitching of the body was considered as positive.
Visual reactivity was derived from the reaction to a dark sheet of paper, brought near to the animal without any physical contact. Turning towards the paper was considered as positive.
For the testing of the righting reflex, the animals were held between the front- and hind legs, turned on their backs and dropped from a height of 30 cm onto the bedding material of the standard arena. A landing on all four legs is considered to be a normal reaction.
The measurement of the forelimb grip strength was determined using a Digital Force Gauge DFIS 100.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes .
Animals were killed by inhalation of 80 % CO2 plus 20 % air and subjected to a necropsy including a gross pathological examination immediately after death on
Day 92: groups K, A, B and C (males).
Day 93: groups K, A, B and C (females).
Day 121: groups KS and CS.
The following organs / tissues were taken from all animals and fixed in Bouin's solution:
gross lesions, tissue masses or tumours
adrenal glands
aorta
brain
caecum
coagulating glands
epididymides
eyes
heart
kidneys
large intestine (colon)
liver
lungs
lymph nodes (mandibular, mesenteric)
oesophagus
ovaries
pancreas
pituitary gland
prostate
rectum
salivary glands
sciatic nerve
seminal vesicles
skeletal muscle (thigh)
skin, mammary glands
small intestine (duodenum, ileum, jejunum), prepared as "Swiss Roll"
spinal cord (cervical, thoracal, lumbar)
spleen
sternum with bone marrow
stomach
testes
thymus
thyroid and parathyroid glands
trachea.
urinary bladder
uterus


ORGAN WEIGHTS
Fresh weights of the following organs were determined of all animals at necropsy:
adrenal glands (both together)
brain
epididymides (both together)
heart
liver
kidneys (both together)
ovaries (both together)
spleen
testes (both together)
thymus

HISTOPATHOLOGY: Yes.
Groups K and C: Histopathological examination was performed in all animals of all fixed organs or tissues listed above.
Groups KS, A, B and CS: No histopathological examination was performed in groups KS, A, B and CS, as there was no indication for test substance related effects found in group C.
The tissue trimming was performed according to "Bahnemann et al.: RITA - Registry of Industrial Toxicology Animal Data - Guides for Organ Sampling and Trimming Procedures in Rats"; Exp.Toxicol.Pathol. 47 (1995), p 247 ff. with the following exceptions:
Not all possible sections, as given in the literature, were actually prepared. One section per organ (in paired organs one of each) was made with the following exceptions:
-Brain (3 sections, one at the optic chiasma, the second at the caudal border of the mammillary body, just posterior to the attachment of the pituitary and the third about 2 mm caudal to the transverse fibres of the pons). Representative regions of the brain, especially cerebrum, cerebellum and pons were included in these sections.
-Spinal cord (three sections, a cervical, a thoracal and a lumbar).
-Liver (two sections).
The small intestine (duodenum, jejunum and ileum) was fixed and trimmed to form two "Swiss Rolls" (one of the cranial and one of the caudal part), according to "Moolenbeek and Ruitenberg: The Swiss Roll, a simple technique for histological studies of rodent intestine"; Lab.Animals 15 (1981), p.57-59.
The trimmed samples of organs or tissues, as described above, were embedded in paraffin. Sections of about 5 ¿m were stained with haematoxylin and eosin (H&E). Evaluation of slides was performed using a light microscope Leica-DMRB.
The frozen liver samples were not subjected to an examination for fatty changes in frozen sections, as there was no indication for alterations noted in the H&E stained sections.
To describe the severity of lesions, the following grades were applied, if appropriate: minimal (1), mild (2), moderate (3), marked (4), severe (5).
The term "focal" together with a higher degree of severity also stands for "multifocal".
Other examinations:
None.
Statistics:
Analysis of variance followed by the Scheffé-test used for all data with means and standard deviations determined, comparison of more than two groups.
t-test used for all data with means and standard deviations determined, for comparison of two groups only.
H-test of Kruskal and Wallis followed by the test of Nemenyi used for counted events with scoring or in cases where the requirements for the analysis of variance were not fulfilled.
Chi2-test used for counted events.
Fisher's exact test used for counted events, if the Chi2-Test was not applicable.
Results were analysed separately for males and females. P = 0.05 was chosen in each test. Two tailed test were used. Groups K and KS and C and CS were treated separately for statistical analysis.
Clinical signs:
effects observed, treatment-related
Mortality:
mortality observed, treatment-related
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Haematological findings:
effects observed, treatment-related
Clinical biochemistry findings:
effects observed, treatment-related
Urinalysis findings:
effects observed, treatment-related
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY
Daily observation: No animal died. The faeces of all dosed animals were stained yellow. As the test substance itself is a yellow dye, the stain of the faeces is attributed to excreted test substance. One single high dosed male was found to be stained on parts of the body surface. The stain of the faeces and of the body's surface are not considered to be toxic effects.
Isolated cases of chromodakryorrhoea, alopecia and one broken incisor lack a dose relationship and are not related to the test substance.
Detailed observations: As in the daily observations, isolated cases of chromodakryorrhoea, alopecia and one broken incisor were noted, but lack a dose relationship and are not related to the test substance.

BODY WEIGHT AND WEIGHT GAIN:
There was no statistically significant difference or dose related trend noted in the body weights of both sexes.
In the body weight gain, isolated terms with significant differences in the mid or high dosed groups, with both higher and lower body weight gains, are considered to be incidental significances.

FOOD CONSUMPTION
There were no noteworthy differences or dose related trends noted in the feed consumption of both sexes. The differences between the groups are too small the give an indication for a toxic effect.

OPHTHALMOSCOPIC EXAMINATION: No indications for ocular lesions were noted at the examinations before the first dosing and at the end of the dosing period. An additional examination at the end of the recovery period was therefore omitted.

HAEMATOLOGY
There were no relevant differences during the dosing period. Small but significant changes were observed at the end of the recovery period: RBC and haemoglobin decreased in males; MCV increased in males; MCHC decreased in females. The differences are minor in dimension, were noted only at the end of the recovery period and lack corresponding effects at the end of the dosing period. Thus they are not regarded as test substance related and may rather be incidental.

CLINICAL CHEMISTRY
Alanin aminotransferase (males) lowered in the mid dosed group and elevated in the high dosed group; cholesterol (females) increased in the mid dosed group; urea (females) decreased in the recovery group.These significant group differences lack a dose related trend. Thus none of the significant group differences is attributed to the test substance.

URINALYSIS
In the females, the low dosed group produced significantly less urine than the controls. As there was no dose relationship in the mid and the high dosed group, this is not related to the test substance.
All dosed groups of the females had a significantly lower urinary pH than the controls, but there was no dose related trend in pH. All mean urine pH data were in a range, commonly seen in control rats. Thus they are not regarded as test substance related.
In the males, there were no significant or otherwise noteworthy group differences in urinalysis results.

NEUROBEHAVIOUR: Neither test substance related findings were made nor were there any significant group differences at the functional observations. All results represent a normal pattern of behaviour and normal reactions of rats of the strain and age examined.
There was no significant group difference in the grip strengths.

ORGAN WEIGHTS
The significantly altered thymus, brain and ovarian weights lack a dose relationship and are considered to be incidental significances.
Significantly elevated liver weights of the high dosed females may be possibly test substance related. As to the lack of any indications for hepatotoxicity, the changes in female liver weights may be part of an adaptive response. There was neither significant difference nor a corresponding trend in the males.

GROSS PATHOLOGY
In the vast majority of all dosed animals, the content of the gastrointestinal tract was found to be stained yellow. This is attributed to the presence of the test substance and not regarded as toxic effect.
Isolated cases of small testes, fatty tissue necrosis in mesenteric fat, ovarian cysts are considered to be parts of the background pathology without relation to the test substance.

HISTOPATHOLOGY:
There was no test substance related alteration noted histopathologically.
Some isolated findings in histopathology are regarded as part of the background pathology, inconspicuous in type and incidence.

OTHER FINDINGS: None.
Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: see 'Remark'
Key result
Critical effects observed:
not specified

Only parameters are listed for which a significant difference between at least one treatment group and the respective control group was observed for at least one sex. All parameters not showing significant differences are omitted from the tables. A significant difference to the respective control group is indicated by a grey background.

Haematology results.

parameter (sex)

low dose

(% of the negative controls)

mid dose

(% of the negative controls)

high dose

(% of the negative controls)

high dose
after recovery
(% of the negative controls)

red blood cell count(males)

100

101

101

97

haemoglobin conc.(males)

100

100

101

97

mean cell volume(males)

100

100

100

102

mean corpuscular haemoglobin conc.(females)

105

109

115

99

Clinical biochemical results.

parameter (sex)

low dose

(% of the negative controls)

mid dose

(% of the negative controls)

high dose

(% of the negative controls)

high dose
after recovery
(% of the negative controls)

alanin aminotransferase(males)

84

83

88

102

cholesterol(females)

84

86

95

96

urea(females)

92

94

97

89

Urinalysis findings.

parameter (sex)

low dose

mid dose

high dose

high dose
after recovery

total amount(females, % of the negative controls)

58

76

85

133

pH(females; control: pH 7.1; control recovery: pH 7.3)

6.4

6.7

6.1

7.3

Organ weights.

parameter (sex)

low dose

(% of the negative controls)

mid dose

(% of the negative controls)

high dose

(% of the negative controls)

high dose
after recovery
(% of the negative controls)

thymus(males, absolute weight)

121

113

114

120

thymus(males, organ weight/body weight ratio)

121

115

114

120

thymus(males, organ weight/brain weight ratio)

120

118

115

120

brain(males, absolute weight)

100

96

99

100

liver(females, absolute weight)

105

105

110

103

liver(females, organ weight/body weight ratio)

104

103

109

99

liver(females, organ weight/brain weight ratio)

104

104

111

100

ovaries(females, absolute weight)

107

92

87

110

Conclusions:
A 90-day toxicity study in Fischer F344 rats was performed with the test item. Groups of 10 male and 10 female rats received 0, 50, 200 and 1000 mg/kg/day by oral gavage in corn oil for 90 days. The study included additional control and high dose groups of 6 males and 6 females each analyzed after a 4-week recovery period. All animals survived until scheduled termination without any toxic signs in life. In haematology, clinical biochemistry and urinalysis some scattered significant differences, altogether without a clear dose response and all only minor in severity, were noted. Post mortem examination including histopathology did not reveal any test substance related toxic alterations. The test substance caused elevated liver weights in the high dosed females. As there were no corresponding histopathological or clinical-biochemical alterations found, the most likely interpretation is an adaptive response by enzyme induction. No parallel trend was present in the males. The effects did not persist until the end of the recovery period. No other test substance-related effect was noted. The No-observed-adverse-effect-level (NOAEL) of the test item was 1000 mg/kg bw/day in both sexes.
Executive summary:

Guidelines applied

EC-Guideline 2001/59/EC, B.26.,
OECD-Guideline 408, 21 September 1998.

Dose levels

Based on the results of a 14-day dose range finding study (which was part of the present study), the dose levels for this 90-day oral gavage study were selected to be 0, 50, 200 and 1000 mg/kg/day.

Study outline

The test substance, formulated in corn oil, was administered daily for 90 days orally by gavage to SPF-bred Fischer F344/DuCrl rats. One control group (plus a control satellite group) and 3 test substance treated groups (plus a high dose satellite group) were tested, each consisting of 10 males and 10 females (6 males and 6 females in the satellite groups).

Evaluated parameters

Chemical analyses of formulations were conducted twice in the in-life phase of the study to assess concentration and homogeneity.

The following parameters were evaluated: Clinical signs (daily plus weekly in more detail), functional observations (terminally), ophthalmoscopy (initial and terminal), body weights and feed consumption (weekly), haematology plus clinical biochemistry plus urinalysis (terminal), necropsy, organ weights and histopathology.

Results

Accuracy and homogeneity of test substance preparations in corn oil were demonstrated by analyses.

All animals survived until scheduled termination without any toxic signs in life. In haematology, clinical biochemistry and urinalysis some scattered significant differences, altogether without a clear dose response and all only minor in severity, were noted. Post mortem examination including histopathology did not reveal any test substance related toxic alterations. Liver weights of the females were elevated with a dose dependency, which may indicate an adaptive response rather than a toxic effect.

A frequently noted yellow stain of the gastrointestinal contents and of the faeces is attributed to the presence of the administered test substance itself and not regarded as a toxic effect.

Discussion and Conclusion

The test substance caused elevated liver weights in the high dosed females. As there were no corresponding histopathological or clinical-biochemical alterations found, the most likely interpretation is an adaptive response by enzyme induction. No parallel trend was present in the males.

No other test substance related effect was noted.

The effects did not persist until the end of the recovery period.

The No-observed-adverse-effect-level (NOAEL) of the test substance was at 1000 mg per kg body weight in both sexes.

Endpoint:
sub-chronic toxicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
For details on endpoint specific justification please see read-across report in section 13 or find a link in cross-reference “assessment report”.
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
assessment report
Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: see 'Remark'
Remarks on result:
other: The reported value refers to the source substance and was not converted to the target substance since (a) no adverse effects were observed and (b) the difference of the molecular weights of the target and the source substance is small (less than 20%).
Key result
Critical effects observed:
not specified
Conclusions:
A 90-day toxicity study in Fischer F344 rats was performed with the test item. Groups of 10 male and 10 female rats received 0, 50, 200 and 1000 mg/kg/day by oral gavage in corn oil for 90 days. The study included additional control and high dose groups of 6 males and 6 females each analyzed after a 4-week recovery period. All animals survived until scheduled termination without any toxic signs in life. In haematology, clinical biochemistry and urinalysis some scattered significant differences, altogether without a clear dose response and all only minor in severity, were noted. Post mortem examination including histopathology did not reveal any test substance related toxic alterations. The test substance caused elevated liver weights in the high dosed females. As there were no corresponding histopathological or clinical-biochemical alterations found, the most likely interpretation is an adaptive response by enzyme induction. No parallel trend was present in the males. The effects did not persist until the end of the recovery period. No other test substance-related effect was noted. The No-observed-adverse-effect-level (NOAEL) of the test item was 1000 mg/kg bw/day in both sexes.
Executive summary:

The study used as source investigated C.I. Pigment Yellow 74. The study results of the source compound were considered applicable to the target compound and were used for classification and labelling acc. to REGULATION (EC) No 1272/2008. Justification and applicability of the read-across approach (structural analogue) is outlined in the read-across report in section 13 or find a link in cross reference “assessment report”.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
reliable

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records

Referenceopen allclose all

Endpoint:
sub-chronic toxicity: inhalation
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
05 July 2021 to 09 June 2022
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
See read across document in chapter 13
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
OECD Guideline 413 (90-Day (Subchronic) Inhalation Toxicity Study
GLP compliance:
yes (incl. QA statement)
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: In-house bred animals
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 7-8 weeks at the time of randomization
- Weight at study initiation: Males: 140.05 g to 158.83 g
Females: 132.98 g to 139.96 g
- Fasting period before study: 16 to 18 hours
- Housing: polysulphonate cage (size: L 430 × B 280 × H 210 mm)
- Diet: Altromin Maintenance diet for rats and mice (manufactured by Altromin Spezialfutter GmbH & Co. KG) (except during restraining and exposure period)
- Water (e.g. ad libitum): yes, Deep bore-well water passed through a reverse osmosis unit
- Acclimation period: Males: 05 July 2021 to 11 July 2021
Females: 12 July 2021 to 18 July 2021



ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.6°C to 23.0°C
- Humidity (%): 44% to 67%
- 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: 05 July 2021 To: 14 April 2022
Route of administration:
inhalation: dust
Type of inhalation exposure:
nose only
Vehicle:
air
Mass median aerodynamic diameter (MMAD):
> 1.8 - <= 2.01 µm
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: flow-past, nose-only dynamic inhalation exposure system supplied by CH Technologies, USA
- Method of holding animals in test chamber: restrainer
- Source and rate of air: compressed air, 20 lt/min
- Method of conditioning air: filtered air through filters
- System of generating particulates/aerosols: dust generator
- Temperature, humidity, pressure in air chamber: 21.5 to 23.6 °C, 51.2% to 59.2%, 20 lt/min
- Air flow rate: 20 lt/min
- Air change rate: 12 to 15 air changes per hour
- Method of particle size determination: gravimetric
- Treatment of exhaust air: NaOH

TEST ATMOSPHERE
- Brief description of analytical method used: no
- Samples taken from breathing zone: yes

Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Actual aerosol concentration was determined by gravimetric method i.e. dividing the mass of test item collected on the filter paper by the volume of air passed through the filter paper and time. The 2.40 L/min critical orifice was used to draw the air from the inhalation chamber for five minutes at animal breathing zone. It was carried out for four times during exposure, initially before putting the animals into the chambers to ensure the chamber concentration and every two hours [i.e. 120 minutes (±15 minutes), 240 minutes (±15 minutes) and 360 minutes (±15 minutes)] after equilibration period for each exposure day.
Duration of treatment / exposure:
Animals were exposed to the test item by inhalation continuously for 6 hours per day, 5 days per week, for a total of 65 actual exposures, after equilibration period of the chamber concentration.
Following the 90 days exposure period, the animals in the recovery groups (Satellite 1 - G1R – S1, G2R – S1, G3R- S1, G4R – S1 and G5R – S1) and (Satellite 2 - G1R- S2, G2R – S2, G3R – S2 G4R – S2 and G5R – S2) were not given any treatment and maintained for 90 days (Satellite 1) and 180 days (Satellite 2) post treatment period and observed for reversibility or persistence of toxic effects
Frequency of treatment:
6 hours per day, 5 days per week
Dose / conc.:
0.03 mg/L air
Remarks:
G5 & G5R
Dose / conc.:
0.01 mg/L air
Remarks:
G4 & G4R
Dose / conc.:
0.003 mg/L air
Remarks:
G3
Dose / conc.:
0.001 mg/L air
Remarks:
G2 &G2R
Dose / conc.:
0 mg/L air
Remarks:
G1 & G1R
No. of animals per sex per dose:
10 M and 10 F
Control animals:
yes, sham-exposed
Details on study design:
- Dose selection rationale: As per sponsor provided doses
- Rationale for animal assignment (if not random): randomisation
- Fasting period before blood sampling for clinical biochemistry: yes, overnight
- Rationale for selecting satellite groups: to observe delayed toxic of test item and recovery
- Post-exposure recovery period in satellite groups: yes
Positive control:
No
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily once
- 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

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: week 13 for G1 and G5, week 26 and 39 for recovery group animals G1R, G5R Satelite-I and G1R, G5R Satelite-II
- Dose groups that were examined: G1, G1R, G5, G5R

HAEMATOLOGY: Yes
- Time schedule for collection of blood: day 90 for main group, day 180 and 270 for recovery group
- Anaesthetic used for blood collection: Yes, isoflurane
- Animals fasted: Yes
- How many animals: 100 animals in main group, 100 animals in recovery group
- Parameters checked in table [No.10] were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: day 90 for main group, day 180 and 270 for recovery group
- Animals fasted: Yes
- How many animals: 100 animals in main group, 100 animals in recovery group
- Parameters checked in table [No.11] were examined.

URINALYSIS: Yes
- Time schedule for collection of urine: day 90 for main group, day 180 and 270 for recovery group
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters checked in table [No. 12] were examined.

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: week 13 for main group, week 26 for satellite 1 group, week 39 for satellite 2 group
- Dose groups that were examined: G1, G1R, G5, G5R
- Battery of functions tested: sensory activity / grip strength / motor activity

BRONCHOALVEOLAR LAVAGE FLUID (BALF): Yes
- Time schedule for analysis: day 90 for main group, day 180 and 270 for recovery group
- Dose groups that were examined: all groups including recovery
- Number of animals: 200
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
Statistics:
SPSS Software version 22
Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
No toxicologically significant treatment-related variations were observed in all the tested dose groups when compared with control group. However, the following statistically significant variations were noted.
In main group males, increase in body weight was noted in G5 from day 71 to day 89 which is considered incidental as no concomitant variation was noted in feed consumption
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
No statistically significant treatment-related variations were observed in all the tested dose groups when compared with control group. However, the following statistically significant variations were noted.
In main group females, decrease in feed consumption was noted in G4 during week 10. The observed change in feed consumption was not accompanied by any changes in mean body weight and hence considered to be incidental
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):
No toxicologically significant treatment related changes in haematology parameters were noted. However, the following statistically significant variations were noted.
In main group males, increase in WBC (G2), RBC (G3, G4 and G5), PT (G2, G3, G4 and G5) and absolute neutrophils (G2, G3, G4 and G5); decrease in MCH (G2, G3, G4 and G5), percent lymphocytes (G3 and G4) and percent basophils (G4) were noted. In main group females, increase in MCV and APTT (G4); decrease in WBC (G3, G4 and G5), Haemoglobin (G4), MCHC (G3, G4 and G5), absolute lymphocytes (G3, G4 and G5) and absolute monocytes (G3) was noted.
In recovery group (S1) males, decrease in Haematocrit (G4R-S1), percent basophils and absolute basolphils (G2R-S1 to G5R-S1), percent lymphocytes (G5R-S1), absolute lymphocytes (G3R-S1 and G5R-S1); increase in percent neutrophils (G3R-S1 and G5R-S1) noted. In females decrease in PT (G4R-S1 and G5R-S1) was noted.
In recovery group (S2) males, decrease in PT (G5R-S2) was noted. In females, increase in WBC and absolute lymphocytes (G2R-S2); decrease in PT (G5R-S2) and APTT (G2R-S2) was noted.
Main: The observed variations in WBC and differential counts at the end of treatment period could be secondary due to exposure of test item and was not associated in any microscopic changes in the marrow and hence considered incidental. Variations noted in MCH and MCHC is also considered incidental in the absence of associated variation in haemoglobin concentration (except haemoglobin concentration of G4F which lacks dose dependency). Increase in PT in main group males was minimal in nature and also in the absence of variation in other coagulation parameter, the noted change in considered incidental. All other changes noted in main groups are considered incidental in the absence of dose dependency anddue to random biological response.
Recovery: At the end of recovery period, few variations in differential counts in males (90-day recovery groups) still persisted which were minimal in nature. All other variations noted at the end of recovery period lacked dose dependency, were random in nature, or were not noted in other sex
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
No toxicologically significant changes were observed in clinical chemistry parameters when compared to the vehicle control group. However, the following statistically significant variations were noted.
In main group, males increase in glucose (G4), creatinine (G3 and G4), albumin (G2, G3, G4 and G5), ALT (G2 and G5), calcium (G3, G4 and G5), chloride (G3, G4 and G5) as well as decrease in ALP and phosphorous (G2, G3, G4 and G5) were noted.
In main group females, increase in creatinine (G2, G3 and G4), triglycerides (G2), albumin (G2, G3, G4 and G5), total bilirubin (G4), sodium (G4 and G5) and Chloride (G3, G4 and G5); decrease in phosphorous (G2, G3, G4 and G5) and Globulin (G2 and G3) was noted.
The noted variations in albumin, ALP, ALT, calcium, chloride were not associated with any macroscopic changes at all the dose levels and no microscopic changes were noted at the high dose group in associated organs. All the other changes noted are considred incidental in the absence of dose dependency and the changes could be due to random physiological variation.
In recovery group (S1) males, increase in triglycerides (G5R-S1), sodium (G2R-S1 to G5R-S1), chloride (G4R-S1 and G5R-S1) were noted; in females increase in glucose and BUN (G2R-S1 and G4R-S1) were noted.
In recovery group (S2) males, increase in potassium (G4R-S2) was noted; in females decrease in urea and BUN (G3R-S2) and increase in ALT (G2R-S2), glucose and potassium (G3R-S2) were noted.
Variations in the recovery group were also considered incidental as they lacked dose dependency patterns
Endocrine findings:
not examined
Urinalysis findings:
effects observed, non-treatment-related
Description (incidence and severity):
No adverse treatment-related effects were observed in urinalysis parameters when compared to the control group. However, in main group females, increase in urine volume (G3) was noted. The noted variation is considered incidental due to its isolated occurance and also lacked dose dependency
Behaviour (functional findings):
effects observed, non-treatment-related
Description (incidence and severity):
There were no adverse effects observed during neurological/functional examination. However, in main group females (G5), decrease in movement counts was noted. This variation is considered incidental due to lack of dose responsiveness and also none of the other neuromuscular parameters were affected
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
No adverse changes were noted in fasting body weights, organ weights and its ratios. However, the following statistically significant variations were noted in organ weights and their ratios.
In main group males, increase in absolute weight of adrenals and kidneys (G5), epididymides and heart (G3, G4 and G5), brain (G3 and G5); decrease in absolute weight of thymus (G5), spleen (G4 and G5); increase in relative weight of heart (G3 and G5), epididymides (G3); decrease in relative weight of thymus (G2, G4 and G5), and spleen (G4 and G5) was noted. In females increase in absolute weight of adrenals and spleen (G5), uterus (G3 and G4), decrease in heart (G3), kidneys (G2 and G3), brain (G3), liver (G3 and G5); increase in realative weight of adrenals and spleen (G5), uterus (G3 and G4); decrease in kidneys (G2) and liver (G5) were noted.
In recovery group (S1) males, increase in relative weight of epididymides (G5R-S1) noted. In females decrease in absolute weight of adrenals (G3R-S1) and liver (G2R-S1) were noted.
In recovery group (S2) males, decrease in absolute weight of adrenals and relative weight of adrenals (G2R-S2) were noted.
All the variations noted in the groups were dose-independent, and were not associated with any macroscopic findings, hence the observed changes in absolute organ weights were considered incidental.
Relative weight changes (relative to brain weight)
In main group males, decrease in relative weight of thymus (G2, G4 and G5), spleen (G4 and G5), testies (G5) and lungs (G2 and G5), decrease in epididymides (G3 and G4) and heart (G4 and G5); in females increase in relative weight of adrenal and spleen (G5) and uterus (G3 and G4); decrease in relative weight of kidneys (G2) and Liver (G2 and G5) was noted.
In recovery group (S1) males, increase in relative weight of epididymides (G5R-S1) was noted. In females decrease in relative weight of adrenals (G3R-S1) and liver (G2R-S1) was noted.
In recovery group (S2) females, decrease in relative weight of kidneys (G3R-S2) was noted.
All variations noted in organ weights were not associated with any macroscopic examination and also no microscopic changes were noted in the high dose main group animals and hence considered incidental
Gross pathological findings:
no effects observed
Neuropathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
There were no test item-related, microscopic findings in the study.
Few of the microscopic findings observed in this study such as ultimobranchial cyst(s) in thyroid gland, epithelial cyst(s)in thymus and all other findings were considered incidental as they occurred randomly across the dose groups including concurrent controls and/or were expected for laboratory rats
Histopathological findings: neoplastic:
no effects observed
Key result
Dose descriptor:
NOAEC
Effect level:
> 30 mg/m³ air (analytical)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
behaviour (functional findings)
body weight and weight gain
clinical biochemistry
clinical signs
food consumption and compound intake
food efficiency
gross pathology
haematology
histopathology: non-neoplastic
immunology
mortality
ophthalmological examination
organ weights and organ / body weight ratios
serum/plasma biochemistry
urinalysis
Key result
Critical effects observed:
no

  • SUMMARY OF CLINICAL SIGNS AND MORTALITY RECORD


                                                                                                                          Refer Appendix 1
































































































































































































Group, Sex & Dose


(mg/L)



No. of


Animals



Clinical Signs of Toxicity



Mortality                                (No. of Incidences/


No. of Animals)



G1, M & 0



10



N



0/10



G2, M & 0.001



10



N



0/10



G3, M & 0.003



10



N



0/10



G4, M & 0.01



10



N



0/10



G5, M & 0.03



10



N



0/10



G1, F & 0



10



N



0/10



G2, F & 0.001



10



N



0/10



G3, F & 0.003



10



N



0/10



G4, F & 0.01



10



N



0/10



G5, F & 0.03



10



N



0/10



G1R - S1, M & 0



05



N



0/05



G2R - S1, M& 0.001



05



N



0/05



G3R - S1, M & 0.003



05



N



0/05



G4R - S1, M & 0.01



05



N



0/05



G5R - S1, M & 0.03



05



N



0/05



G1R - S1, F & 0



05



N



0/05



G2R - S1, F & 0.001



05



N



0/05



G3R - S1, F & 0.003



05



N



0/05



G4R - S1, F & 0.01



05



N



0/05



G5R - S1, F & 0.03



05



N



0/05



G1R – S2, M & 0



05



N



0/05



G2R – S2, M& 0.001



05



N



0/05



G3R – S2, M & 0.003



05



N



0/05



G4R – S2, M & 0.01



05



N



0/05



G5R – S2, M & 0.03



05



N



0/05



G1R – S2, F & 0



05



N



0/05



G2R – S2, F & 0.001



05



N



0/05



G3R – S2, F & 0.003



05



N



0/05



G4R – S2, F & 0.01



05



N



0/05



G5R – S2, F & 0.03



05



N



0/05



M: Male; F: Female; N: Normal; R: Recovery; S1: Satellite 1; S2: Satellite 2.


 


 



  • SUMMARY OF BODY WEIGHTS (g) RECORD


  Refer Appendix 2








































































































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Body Weight (g) on Days



1



5



8



12



15



19



22



26



29



G1, M & 0



Mean



175.96



181.61



193.88



204.16



215.88



225.54



237.96



248.86



257.86



±SD



8.76



9.75



11.15



11.52



12.06



12.44



13.92



14.93



14.76



n



10



10



10



10



10



10



10



10



10



G2, M & 0.001



Mean



178.67



185.75



199.35



210.00



222.44



233.11



247.09



257.79



267.58



±SD



9.57



9.28



9.79



9.57



9.18



9.16



10.14



9.28



9.62



n



10



10



10



10



10



10



10



10



10



G3, M & 0.003



Mean



179.30



185.56



196.82



206.48



217.41



227.67



241.18



252.16



261.28



±SD



9.05



8.71



10.45



10.61



12.95



13.88



15.10



16.73



18.34



n



10



10



10



10



10



10



10



10



10



G4, M & 0.01



Mean



179.76



186.47



200.04



210.60



225.07



234.63



248.74



260.15



269.78



±SD



10.27



9.91



10.79



10.96



10.20



10.24



10.49



11.25



11.58



n



10



10



10



10



10



10



10



10



10



G5, M & 0.03



Mean



178.42



184.59



199.02



209.71



222.87



232.04



246.51



257.09



266.58



±SD



8.24



9.02



8.82



8.17



9.39



9.26



10.45



10.73



10.62



n



10



10



10



10



10



10



10



10



10



G1, F & 0



Mean



169.17



172.82



178.30



183.68



190.80



195.57



200.33



203.64



208.25



±SD



7.40



7.37



8.53



10.28



10.01



10.66



11.55



11.27



11.07



n



10



10



10



10



10



10



10



10



10



G2, F & 0.001



Mean



167.60



171.44



178.13



183.92



191.08



195.60



200.77



204.55



209.39



±SD



6.08



6.53



6.93



8.15



10.18



10.96



13.55



15.67



16.01



n



10



10



10



10



10



10



10



10



10



G3, F & 0.003



Mean



168.11



171.34



176.54



182.06



188.93



193.63



198.66



202.57



207.79



±SD



4.20



3.78



5.14



5.78



7.08



8.92



9.67



9.91



9.97



n



10



10



10



10



10



10



10



10



10



G4, F & 0.01



Mean



168.05



171.50



176.61



181.97



189.28



194.60



199.69



203.74



209.20



±SD



4.80



5.82



7.47



7.77



10.19



12.28



13.18



12.47



12.39



n



10



10



10



10



10



10



10



10



10



G5, F & 0.03



Mean



169.10



172.54



177.88



183.21



190.25



194.49



200.14



204.40



209.04



±SD



5.71



6.05



7.45



8.76



10.94



12.19



11.43



13.49



13.71



n



10



10



10



10



10



10



10



10



10



 M: Male; F: Female; SD: Standard Deviation; n: Number of animals.


 



  • TABLE 2 (Contd...). SUMMARY OF BODY WEIGHTS (g) RECORD


  Refer Appendix 2








































































































































































































































































































































































































 Group, Sex & Dose (mg/L)


 

Body Weight (g) on Days



36



43



50



57



64



71



78



85



89



G1, M & 0



Mean



268.53



286.06



302.17



316.94



329.47



358.31



373.62



389.40



396.27



±SD



17.53



18.93



20.67



23.41



23.63



21.47



20.30



21.73



22.30



n



10



10



10



10



10



10



10



10



10



G2, M & 0.001



Mean



278.37



295.90



312.50



329.74



343.41



373.68



388.80



405.15



412.39



±SD



11.14



14.06



19.56



24.75



24.33



22.30



21.11



25.46



24.24



n



10



10



10



10



10



10



10



10



10



G3, M & 0.003



Mean



271.47



290.02



308.39



321.40



335.17



366.09



381.88



396.39



404.29



±SD



18.38



22.47



27.82



30.11



30.68



27.66



27.61



27.22



27.46



n



10



10



10



10



10



10



10



10



10



G4, M & 0.01



Mean



281.18



298.07



318.69



332.01



346.44



374.79



390.05



404.33



413.99



±SD



11.75



9.43



11.62



12.58



12.54



13.61



13.45



13.47



12.21



n



10



10



10



10



10



10



10



10



10



G5, M & 0.03



Mean



278.20



297.51



320.45



329.47



353.83



386.35*



402.21*



418.61*



425.77*



±SD



11.15



13.63



21.28



37.61



22.13



21.27



20.72



20.80



20.70



n



10



10



10



10



10



10



10



10



10



G1, F & 0



Mean



218.52



225.49



231.32



238.59



244.95



255.28



261.20



267.37



272.80



±SD



11.93



11.81



11.31



11.02



11.48



10.91



10.77



11.29



11.18



n



10



10



10



10



10



10



10



10



10



G2, F & 0.001



Mean



217.34



223.70



231.65



237.65



243.84



259.86



262.49



269.14



275.20



±SD



17.44



17.52



16.72



16.55



16.70



18.52



18.70



18.25



18.22



n



10



10



10



10



10



10



10



10



10



G3, F & 0.003



Mean



216.66



222.43



228.13



235.11



242.70



253.07



259.73



266.14



271.06



±SD



9.56



7.55



7.43



8.08



7.95



7.31



6.65



6.53



6.32



n



10



10



10



10



10



10



10



10



10



G4, F & 0.01



Mean



216.09



223.02



230.72



236.68



243.51



256.30



262.37



269.03



274.17



±SD



13.00



12.51



9.24



8.99



8.51



7.94



8.25



8.34



8.89



n



10



10



10



10



10



10



10



10



10



G5, F & 0.03



Mean



216.56



222.52



228.31



234.70



241.05



253.04



259.25



265.37



271.24



±SD



13.13



14.26



14.61



14.74



14.71



14.90



14.12



13.77



13.56



n



10



10



10



10



10



10



10



10



10



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; *: Statistically significant (p<0.05)


 



  • TABLE 2 (Contd...). SUMMARY OF BODY WEIGHTS (g) RECORD


  Refer Appendix 2








































































































































































































































































































































































































 Group, Sex & Dose (mg/L)


 

Body Weight (g) on Days



1



5



8



12



15



19



22



26



29



G1R-S1, M & 0



Mean



180.49



186.10



200.07



209.29



221.44



228.07



234.45



246.19



255.57



±SD



11.74



12.27



13.20



12.69



14.60



14.81



15.23



16.35



16.17



n



5



5



5



5



5



5



5



5



5



G2R-S1, M & 0.001



Mean



183.70



189.91



201.46



211.27



224.05



230.11



237.14



247.06



255.80



±SD



12.15



11.37



14.70



14.31



15.65



16.19



18.23



16.75



16.10



n



5



5



5



5



5



5



5



5



5



G3R-S1, M & 0.003



Mean



180.10



184.76



199.54



209.36



222.04



228.01



235.83



248.19



258.70



±SD



11.20



10.73



12.74



12.85



11.64



12.09



11.36



11.80



13.12



n



5



5



5



5



5



5



5



5



5



G4R-S1, M & 0.01



Mean



180.66



185.81



199.74



210.55



222.34



229.16



236.95



248.76



257.32



±SD



10.73



10.15



11.18



10.43



9.48



9.72



10.40



10.42



8.85



n



5



5



5



5



5



5



5



5



5



G5R-S1, M & 0.03



Mean



179.27



184.58



198.95



209.56



222.12



228.07



235.87



247.43



258.14



±SD



7.75



8.46



10.09



10.20



10.71



10.22



10.63



11.52



11.85



n



5



5



5



5



5



5



5



5



5



G1R-S1, F & 0



Mean



168.63



171.56



179.12



185.86



190.92



195.58



199.96



203.83



207.77



±SD



6.77



7.43



5.63



4.87



5.07



3.05



3.52



3.80



4.42



n



5



5



5



5



5



5



5



5



5



G2R-S1, F & 0.001



Mean



170.49



173.82



178.83



186.21



191.46



196.17



200.47



204.66



208.77



±SD



5.57



5.31



3.64



4.82



5.68



5.74



6.69



7.92



9.24



n



5



5



5



5



5



5



5



5



5



G3R-S1, F & 0.003



Mean



170.52



174.16



182.18



188.83



194.93



199.80



204.01



208.62



212.99



±SD



4.84



3.77



6.41



6.40



7.12



6.18



5.34



6.03



6.53



n



5



5



5



5



5



5



5



5



5



G4R-S1, F & 0.01



Mean



171.44



174.65



181.56



188.18



193.92



198.51



202.72



207.83



212.28



±SD



2.54



1.34



3.37



4.83



4.32



4.17



3.62



2.40



2.51



n



5



5



5



5



5



5



5



5



5



G5R-S1, F & 0.03



Mean



169.40



172.28



179.83



186.50



192.10



196.72



200.83



205.26



209.72



±SD



4.89



5.44



6.96



6.96



7.93



7.21



7.16



8.33



8.20



n



5



5



5



5



5



5



5



5



5



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; R: Recovery; S1: Satellite 1.


 


TABLE 2 (Contd...). SUMMARY OF BODY WEIGHTS (g) RECORD


  Refer Appendix 2








































































































































































































































































































































































































 Group, Sex & Dose (mg/L)


 

Body Weight (g) on Days



36



43



50



57



64



71



78



85



92



G1R-S1, M & 0



Mean



264.87



286.34



306.62



322.94



338.25



365.89



383.08



396.56



409.85



±SD



16.21



28.24



31.99



29.52



29.47



27.26



27.40



25.86



26.26



n



5



5



5



5



5



5



5



5



5



G2R-S1, M & 0.001



Mean



269.43



296.61



316.47



332.01



343.98



373.31



389.80



399.77



413.69



±SD



15.79



13.30



18.81



18.05



18.05



18.09



17.03



18.25



18.56



n



5



5



5



5



5



5



5



5



5



G3R-S1, M & 0.003



Mean



270.43



295.54



315.92



327.24



338.40



350.72



368.98



379.32



392.67



±SD



11.78



11.05



11.82



11.54



11.97



17.00



16.56



17.10



16.91



n



5



5



5



5



5



5



5



5



5



G4R-S1, M & 0.01



Mean



269.80



300.31



316.79



333.06



345.53



372.37



388.88



402.30



416.82



±SD



7.59



7.68



11.90



15.79



14.94



16.34



16.15



16.51



15.85



n



5



5



5



5



5



5



5



5



5



G5R-S1, M & 0.03



Mean



269.00



290.70



310.63



325.42



335.57



349.87



366.79



378.04



392.66



±SD



11.59



15.42



22.82



27.49



27.47



29.94



29.20



29.37



29.72



n



5



5



5



5



5



5



5



5



5



G1R-S1, F & 0



Mean



224.93



230.83



238.35



244.45



251.05



262.76



269.60



276.36



283.29



±SD



4.12



6.60



8.08



7.70



7.70



8.42



9.34



8.56



8.91



n



5



5



5



5



5



5



5



5



5



G2R-S1, F & 0.001



Mean



223.06



231.28



237.06



243.33



249.34



260.60



263.09



268.95



276.30



±SD



12.00



14.27



13.83



14.49



14.36



13.26



11.28



11.06



11.09



n



5



5



5



5



5



5



5



5



5



G3R-S1, F & 0.003



Mean



231.78



239.41



244.42



250.87



256.61



266.80



274.51



280.77



287.53



±SD



7.57



11.30



10.87



10.81



12.00



10.48



11.42



11.80



11.31



n



5



5



5



5



5



5



5



5



5



G4R-S1, F & 0.01



Mean



228.32



237.20



243.45



250.89



258.14



267.74



274.22



279.59



286.74



±SD



7.52



8.52



11.13



11.08



10.54



10.23



10.63



11.01



10.88



n



5



5



5



5



5



5



5



5



5



G5R-S1, F & 0.03



Mean



227.06



231.11



237.22



245.44



250.61



261.49



267.84



274.99



280.89



±SD



11.68



12.14



13.11



12.88



12.71



11.19



11.71



11.25



11.26



n



5



5



5



5



5



5



5



5



5



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; R: Recovery; S1: Satellite 1.


 



  • TABLE 2 (Contd...). SUMMARY OF BODY WEIGHTS (g) RECORD


  Refer Appendix 2










































































































































































































































































































































 Group, Sex & Dose (mg/L)


 

Body Weight (g) on Days



99



106



113



120



127



134



141



G1R-S1, M & 0



Mean



428.38



440.04



451.46



464.13



477.22



490.12



494.91



±SD



23.46



23.34



24.54



23.33



23.46



23.29



23.41



n



5



5



5



5



5



5



5



G2R-S1, M & 0.001



Mean



435.01



447.84



461.87



475.58



488.78



503.18



510.14



±SD



18.84



19.63



18.25



15.23



14.35



13.34



13.86



n



5



5



5



5



5



5



5



G3R-S1, M & 0.003



Mean



407.88



422.83



438.00



452.07



466.09



478.19



485.48



±SD



18.47



19.48



19.71



19.64



19.96



19.26



18.59



n



5



5



5



5



5



5



5



G4R-S1, M & 0.01



Mean



431.07



443.59



457.47



471.96



487.87



500.52



507.79



±SD



16.26



17.26



18.86



17.56



17.52



16.97



17.29



n



5



5



5



5



5



5



5



G5R-S1, M & 0.03



Mean



408.47



422.87



437.51



451.83



466.70



479.05



486.15



±SD



29.71



32.00



31.65



32.07



32.41



33.18



32.74



n



5



5



5



5



5



5



5



G1R-S1, F & 0



Mean



294.00



302.68



311.54



324.00



331.18



335.15



343.13



±SD



8.12



6.73



7.86



7.70



8.03



8.23



8.89



n



5



5



5



5



5



5



5



G2R-S1, F & 0.001



Mean



284.60



291.16



302.64



314.60



321.71



326.46



334.08



±SD



12.04



12.46



11.99



11.89



11.80



12.00



11.89



n



5



5



5



5



5



5



5



G3R-S1, F & 0.003



Mean



294.63



302.31



311.20



321.97



329.55



334.45



342.66



±SD



12.55



12.56



11.48



11.54



11.23



11.35



11.99



n



5



5



5



5



5



5



5



G4R-S1, F & 0.01



Mean



292.87



301.44



311.19



323.52



330.79



335.44



343.69



±SD



10.81



10.20



9.38



9.43



10.12



10.48



10.51



n



5



5



5



5



5



5



5



G5R-S1, F & 0.03



Mean



286.63



293.80



304.53



315.66



322.10



327.35



334.75



±SD



10.95



10.78



12.14



12.22



11.42



11.44



11.39



n



5



5



5



5



5



5



5



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; R: Recovery; S1: Satellite 1.


 


 



  • TABLE 2 (Contd...). SUMMARY OF BODY WEIGHTS (g) RECORD


  Refer Appendix 2











































































































































































































































































































 Group, Sex & Dose (mg/L)


 

Body Weight (g) on Days



148



155



162



169



176



179



G1R-S1, M & 0



Mean



508.67



520.05



534.27



547.45



560.67



571.20



±SD



23.03



24.29



23.96



23.92



23.93



22.03



n



5



5



5



5



5



5



G2R-S1, M & 0.001



Mean



523.17



536.93



550.74



564.28



577.34



587.83



±SD



13.22



13.43



13.92



12.96



12.20



10.38



n



5



5



5



5



5



5



G3R-S1, M & 0.003



Mean



499.32



512.50



527.45



541.63



554.86



562.77



±SD



18.88



18.71



18.40



17.76



17.72



18.01



n



5



5



5



5



5



5



G4R-S1, M & 0.01



Mean



520.35



533.83



547.51



561.61



574.97



582.76



±SD



17.26



16.85



16.14



16.13



16.50



16.13



n



5



5



5



5



5



5



G5R-S1, M & 0.03



Mean



500.18



514.24



527.48



540.70



555.07



565.03



±SD



32.55



32.16



32.20



31.75



31.85



31.18



n



5



5



5



5



5



5



G1R-S1, F & 0



Mean



352.93



363.40



371.02



380.84



410.27



416.00



±SD



9.71



9.84



9.80



10.06



48.80



47.80



n



5



5



5



5



5



5



G2R-S1, F & 0.001



Mean



346.20



357.36



367.19



377.39



386.46



391.64



±SD



12.20



14.50



15.75



15.18



15.55



15.56



n



5



5



5



5



5



5



G3R-S1, F & 0.003



Mean



353.41



363.06



372.31



382.28



390.97



397.44



±SD



11.96



12.02



12.62



11.93



12.32



11.38



n



5



5



5



5



5



5



G4R-S1, F & 0.01



Mean



355.56



368.54



378.84



388.56



397.86



403.29



±SD



10.01



10.61



10.93



10.76



9.98



9.82



n



5



5



5



5



5



5



G5R-S1, F & 0.03



Mean



345.09



355.32



365.84



372.00



381.19



386.60



±SD



11.40



11.45



11.03



14.69



14.07



13.06



n



5



5



5



5



5



5



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; R: Recovery; S1: Satellite 1.


 


 



  • TABLE 2 (Contd...). SUMMARY OF BODY WEIGHTS (g) RECORD


  Refer Appendix 2








































































































































































































































































































































































































 Group, Sex & Dose (mg/L)


 

Body Weight (g) on Days



1



5



8



12



15



19



22



26



29



G1R-S2, M & 0



Mean



179.28



184.72



201.17



211.04



223.26



229.24



236.49



247.31



257.85



±SD



8.30



8.16



7.99



7.33



8.42



10.16



8.91



8.45



9.23



n



5



5



5



5



5



5



5



5



5



G2R-S2, M & 0.001



Mean



179.04



187.53



202.01



212.11



227.04



233.64



240.58



252.43



260.78



±SD



7.16



7.00



8.03



7.35



5.35



5.16



6.53



9.25



5.66



n



5



5



5



5



5



5



5



5



5



G3R-S2, M & 0.003



Mean



179.63



185.00



200.25



211.05



224.21



230.41



237.83



249.15



259.06



±SD



7.12



5.75



6.92



6.62



8.53



8.79



9.29



10.06



10.75



n



5



5



5



5



5



5



5



5



5



G4R-S2, M & 0.01



Mean



179.33



186.27



200.91



210.55



225.22



229.65



236.76



247.95



257.97



±SD



7.66



7.32



8.86



8.68



9.79



10.36



8.99



10.54



11.36



n



5



5



5



5



5



5



5



5



5



G5R-S2, M & 0.03



Mean



182.11



188.27



203.61



214.95



227.25



233.52



241.09



253.24



263.84



±SD



4.03



3.47



5.55



4.80



5.78



5.44



4.53



3.81



3.66



n



5



5



5



5



5



5



5



5



5



G1R-S2, F & 0



Mean



172.09



175.16



181.36



188.23



192.40



196.98



201.73



206.82



211.16



±SD



6.13



4.64



2.84



3.98



5.27



6.71



6.63



7.21



5.89



n



5



5



5



5



5



5



5



5



5



G2R-S2, F & 0.001



Mean



172.84



176.09



182.08



188.73



194.03



199.21



203.67



208.56



212.37



±SD



2.60



2.16



4.23



7.83



7.42



6.90



6.82



7.24



8.26



n



5



5



5



5



5



5



5



5



5



G3R-S2, F & 0.003



Mean



173.23



176.14



182.70



190.49



196.51



201.58



207.96



212.86



216.96



±SD



7.10



7.72



7.66



6.73



6.92



6.61



8.67



9.70



9.74



n



5



5



5



5



5



5



5



5



5



G4R-S2, F & 0.01



Mean



171.19



174.51



181.38



188.55



193.67



198.94



203.39



208.43



213.14



±SD



8.21



7.43



7.38



11.71



12.25



11.75



11.25



11.74



12.13



n



5



5



5



5



5



5



5



5



5



G5R-S2, F & 0.03



Mean



171.08



174.81



180.97



188.31



193.60



198.63



203.09



208.28



213.56



±SD



2.71



3.76



5.37



7.42



8.19



7.46



6.90



6.51



6.72



n



5



5



5



5



5



5



5



5



5



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; R: Recovery; S2: Satellite 2.


 


 



  • TABLE 2 (Contd...). SUMMARY OF BODY WEIGHTS (g) RECORD


  Refer Appendix 2








































































































































































































































































































































































































 Group, Sex & Dose (mg/L)


 

Body Weight (g) on Days



36



43



50



57



64



71



78



85



92



G1R-S2, M & 0



Mean



271.10



301.04



320.25



334.60



346.29



373.49



388.79



402.28



416.22



±SD



6.83



9.34



16.13



19.17



19.48



21.32



20.64



19.74



19.73



n



5



5



5



5



5



5



5



5



5



G2R-S2, M & 0.001



Mean



272.03



299.98



322.31



333.61



349.06



381.25



395.57



409.07



423.55



±SD



5.76



11.90



18.93



20.97



23.19



19.09



18.00



16.77



16.16



n



5



5



5



5



5



5



5



5



5



G3R-S2, M & 0.003



Mean



270.06



298.66



323.79



342.98



354.24



365.11



378.52



391.07



405.72



±SD



10.09



13.52



20.88



27.14



27.43



26.92



26.27



28.80



28.41



n



5



5



5



5



5



5



5



5



5



G4R-S2, M & 0.01



Mean



268.54



300.05



324.75



342.32



356.01



387.11



400.56



412.72



426.71



±SD



10.94



14.96



21.93



27.34



27.59



24.40



24.02



25.69



25.77



n



5



5



5



5



5



5



5



5



5



G5R-S2, M & 0.03



Mean



273.21



305.37



329.30



344.09



355.54



370.77



382.32



393.97



408.71



±SD



4.66



8.26



18.34



21.55



21.20



26.02



24.94



26.24



27.03



n



5



5



5



5



5



5



5



5



5



G1R-S2, F & 0



Mean



224.14



230.23



237.04



242.91



249.50



261.98



268.45



274.38



282.37



±SD



14.02



16.35



17.25



16.17



15.94



16.18



15.91



15.85



15.63



n



5



5



5



5



5



5



5



5



5



G2R-S2, F & 0.001



Mean



228.70



238.88



245.11



250.03



257.49



267.56



272.58



278.59



284.99



±SD



9.05



12.60



14.47



14.99



14.01



14.74



14.81



15.33



15.38



n



5



5



5



5



5



5



5



5



5



G3R-S2, F & 0.003



Mean



236.89



244.50



250.17



257.48



263.56



274.86



282.38



288.94



294.92



±SD



7.90



5.28



5.89



4.45



4.49



5.00



4.97



4.73



5.00



n



5



5



5



5



5



5



5



5



5



G4R-S2, F & 0.01



Mean



227.39



236.81



246.57



253.58



259.49



270.85



277.65



283.32



290.04



±SD



14.34



17.01



19.32



18.15



17.74



18.25



18.41



18.74



18.84



n



5



5



5



5



5



5



5



5



5



G5R-S2, F & 0.03



Mean



228.77



236.80



241.70



249.02



254.43



264.79



270.83



277.88



285.58



±SD



13.34



17.64



17.78



18.75



18.72



18.51



18.39



18.40



17.45



n



5



5



5



5



5



5



5



5



5



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; R: Recovery; S2: Satellite 2.


 


TABLE 2 (Contd...). SUMMARY OF BODY WEIGHTS (g) RECORD


  Refer Appendix 2








































































































































































































































































































































































































 Group, Sex & Dose (mg/L)


 

Body Weight (g) on Days



99



106



113



120



127



134



141



148



155



G1R-S2, M & 0



Mean



432.45



446.43



464.36



479.38



494.05



506.55



511.83



525.83



539.87



±SD



19.55



19.88



17.67



16.90



17.35



17.34



17.52



16.55



16.96



n



5



5



5



5



5



5



5



5



5



G2R-S2, M & 0.001



Mean



441.48



456.80



474.38



488.89



502.38



515.51



522.52



537.36



550.70



±SD



12.04



15.20



13.60



13.26



12.87



12.02



13.11



10.09



10.49



n



5



5



5



5



5



5



5



5



5



G3R-S2, M & 0.003



Mean



422.25



436.54



454.46



470.10



485.35



497.74



503.93



517.69



531.64



±SD



28.22



28.56



28.97



25.64



26.43



27.01



26.30



26.86



26.84



n



5



5



5



5



5



5



5



5



5



G4R-S2, M & 0.01



Mean



442.57



459.75



474.48



489.33



503.09



515.98



522.97



537.06



551.99



±SD



25.77



26.82



24.92



22.45



22.15



21.28



21.02



19.75



20.81



n



5



5



5



5



5



5



5



5



5



G5R-S2, M & 0.03



Mean



427.54



443.78



460.74



475.19



489.32



501.46



508.99



521.90



536.69



±SD



26.94



26.19



25.30



22.35



22.95



22.95



23.10



23.12



22.40



n



5



5



5



5



5



5



5



5



5



G1R-S2, F & 0



Mean



288.53



296.49



306.59



319.45



326.87



331.56



339.05



351.14



362.65



±SD



15.42



14.16



15.03



15.61



15.02



14.71



14.31



14.16



13.91



n



5



5



5



5



5



5



5



5



5



G2R-S2, F & 0.001



Mean



292.47



298.98



308.76



320.40



328.06



332.62



339.90



351.74



364.02



±SD



14.81



14.42



13.49



12.87



13.13



13.74



13.45



12.82



13.69



n



5



5



5



5



5



5



5



5



5



G3R-S2, F & 0.003



Mean



301.18



308.09



318.45



329.46



336.54



341.12



348.84



359.40



371.43



±SD



5.87



5.65



5.63



5.31



6.02



5.66



6.14



7.45



8.07



n



5



5



5



5



5



5



5



5



5



G4R-S2, F & 0.01



Mean



296.12



303.00



313.06



325.23



332.03



336.93



343.85



356.74



366.91



±SD



19.21



18.75



17.74



18.84



19.23



19.39



18.70



19.28



19.22



n



5



5



5



5



5



5



5



5



5



G5R-S2, F & 0.03



Mean



294.01



301.29



311.23



323.82



329.99



334.97



341.24



354.07



366.58



±SD



16.61



16.74



14.76



14.14



13.78



13.98



14.38



13.79



13.49



n



5



5



5



5



5



5



5



5



5



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; R: Recovery; S2: Satellite 2.


 


TABLE 2 (Contd...). SUMMARY OF BODY WEIGHTS (g) RECORD


  Refer Appendix 2








































































































































































































































































































































































































 Group, Sex & Dose (mg/L)


 

Body Weight (g) on Days



162



169



176



183



190



197



204



211



218



G1R-S2, M & 0



Mean



554.40



569.11



582.97



597.28



610.38



623.56



636.28



649.14



661.36



±SD



16.56



16.18



14.38



14.48



13.39



13.06



13.43



13.52



12.83



n



5



5



5



5



5



5



5



5



5



G2R-S2, M & 0.001



Mean



565.04



578.61



594.12



608.48



620.84



632.69



646.26



656.02



667.31



±SD



10.21



10.55



11.30



9.97



9.65



9.14



9.50



8.48



8.55



n



5



5



5



5



5



5



5



5



5



G3R-S2, M & 0.003



Mean



545.49



558.03



571.33



584.94



597.66



609.25



622.76



633.09



645.43



±SD



26.94



26.46



26.44



27.16



27.34



27.18



27.06



30.11



30.15



n



5



5



5



5



5



5



5



5



5



G4R-S2, M & 0.01



Mean



565.14



578.49



592.92



606.93



620.12



632.13



645.17



658.33



669.94



±SD



20.14



19.34



21.01



20.68



19.43



19.29



19.52



19.20



19.54



n



5



5



5



5



5



5



5



5



5



G5R-S2, M & 0.03



Mean



550.21



564.08



579.68



595.29



607.01



619.28



630.20



643.30



655.39



±SD



22.23



22.46



21.69



23.05



22.40



22.66



22.04



21.58



22.70



n



5



5



5



5



5



5



5



5



5



G1R-S2, F & 0



Mean



373.29



381.72



390.40



398.88



408.60



416.14



423.65



431.03



437.52



±SD



12.62



12.15



11.78



11.65



12.03



11.52



11.26



11.38



11.69



n



5



5



5



5



5



5



5



5



5



G2R-S2, F & 0.001



Mean



359.58



383.56



393.27



401.36



409.62



417.01



425.21



433.01



438.99



±SD



35.96



14.40



13.63



14.28



13.50



13.57



12.76



12.74



13.56



n



5



5



5



5



5



5



5



5



5



G3R-S2, F & 0.003



Mean



382.10



390.46



398.54



406.48



415.37



422.73



430.36



437.84



444.17



±SD



8.59



9.21



9.14



8.75



8.69



8.51



8.29



8.48



8.60



n



5



5



5



5



5



5



5



5



5



G4R-S2, F & 0.01



Mean



374.54



382.74



391.05



399.53



407.99



414.07



422.13



429.85



435.78



±SD



19.37



19.42



18.44



18.79



19.18



18.90



18.86



18.78



19.09



n



5



5



5



5



5



5



5



5



5



G5R-S2, F & 0.03



Mean



374.22



382.08



391.36



399.68



408.14



414.69



421.72



429.76



436.57



±SD



13.31



13.39



13.46



13.74



14.31



13.99



13.68



13.13



12.85



n



5



5



5



5



5



5



5



5



5



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; R: Recovery; S2: Satellite 2.


 


TABLE 2 (Contd...). SUMMARY OF BODY WEIGHTS (g) RECORD


  Refer Appendix 2









































































































































































































































































































































































 Group, Sex & Dose (mg/L)


 

Body Weight (g) on Days



225



232



239



246



253



260



267



269



G1R-S2, M & 0



Mean



670.16



676.42



682.13



690.05



698.99



705.18



711.46



714.99



±SD



12.52



13.26



12.29



8.91



8.51



9.41



8.61



8.63



n



5



5



5



5



5



5



5



5



G2R-S2, M & 0.001



Mean



676.95



683.22



686.79



694.51



705.64



713.39



718.98



722.65



±SD



8.31



8.47



7.72



6.80



7.25



7.35



7.99



8.50



n



5



5



5



5



5



5



5



5



G3R-S2, M & 0.003



Mean



649.71



655.47



660.17



674.95



683.29



690.89



696.75



700.28



±SD



32.62



31.85



30.68



32.39



32.89



32.26



32.00



31.66



n



5



5



5



5



5



5



5



5



G4R-S2, M & 0.01



Mean



678.39



683.60



688.38



699.77



709.67



717.38



723.64



727.23



±SD



18.40



17.93



18.27



21.94



20.36



19.38



20.52



20.56



n



5



5



5



5



5



5



5



5



G5R-S2, M & 0.03



Mean



663.55



669.06



672.69



687.85



694.80



703.04



708.51



711.55



±SD



22.88



22.48



22.56



22.59



23.60



23.69



23.78



23.51



n



5



5



5



5



5



5



5



5



G1R-S2, F & 0



Mean



441.23



444.52



453.91



458.56



463.20



467.56



472.74



475.05



±SD



11.83



11.70



13.20



13.38



13.83



13.86



13.38



13.52



n



5



5



5



5



5



5



5



5



G2R-S2, F & 0.001



Mean



443.15



447.97



457.44



462.94



467.80



471.36



476.93



480.06



±SD



13.66



13.98



14.92



14.97



14.69



14.34



13.57



13.32



n



5



5



5



5



5



5



5



5



G3R-S2, F & 0.003



Mean



447.35



452.66



468.23



473.10



477.21



480.62



485.62



488.14



±SD



8.85



9.04



13.02



13.42



13.35



13.79



13.69



13.98



n



5



5



5



5



5



5



5



5



G4R-S2, F & 0.01



Mean



439.44



443.30



461.63



467.92



472.64



476.62



481.05



483.40



±SD



19.28



18.97



17.70



16.50



17.00



17.18



16.87



16.38



n



5



5



5



5



5



5



5



5



G5R-S2, F & 0.03



Mean



440.20



444.66



458.80



462.97



468.78



472.67



477.51



480.03



±SD



12.50



12.69



12.04



11.63



10.96



10.94



11.16



11.64



n



5



5



5



5



5



5



5



5



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; R: Recovery; S2: Satellite 2.


 


 



  • SUMMARY OF PERCENT CHANGE IN BODY WEIGHT WITH RESPECT TO DAY 1 RECORD


Refer Appendix 3









































































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Percent Change in Body Weight (%) during Days



1 to 5



1 to 8



1 to 12



1 to 15



1 to 19



1 to 22



1 to 26



1 to 29



G1, M & 0



Mean



3.20



10.16



16.01



22.69



28.18



35.23



41.46



46.62



±SD



1.17



1.98



2.46



3.08



3.26



4.16



5.70



6.59



n



10



10



10



10



10



10



10



10



G2, M & 0.001



Mean



3.98



11.60



17.59



24.58



30.58



38.41



44.47



49.98



±SD



0.71



0.82



1.45



2.11



2.66



3.52



5.13



6.03



n



10



10



10



10



10



10



10



10



G3, M & 0.003



Mean



3.52



9.77



15.17



21.23



26.95



34.49



40.59



45.67



±SD



1.13



1.48



1.82



2.69



3.42



4.18



4.73



5.92



n



10



10



10



10



10



10



10



10



G4, M & 0.01



Mean



3.76



11.31



17.20



25.31



30.65



38.52



44.90



50.27



±SD



1.03



1.72



2.07



3.44



3.97



4.29



5.37



5.71



n



10



10



10



10



10



10



10



10



G5, M & 0.03



Mean



3.46



11.58



17.60



24.98



30.14



38.28



44.23



49.58



±SD



1.38



2.37



2.53



3.59



3.87



5.28



5.92



6.75



n



10



10



10



10



10



10



10



10



G1, F & 0



Mean



2.16



5.39



8.55



12.78



15.62



18.41



20.39



23.13



±SD



0.76



1.77



2.64



3.15



4.14



4.18



4.43



4.49



n



10



10



10



10



10



10



10



10



G2, F & 0.001



Mean



2.29



6.28



9.74



14.01



16.73



19.81



22.06



24.94



±SD



1.11



1.34



2.83



4.48



5.77



7.38



8.53



8.66



n



10



10



10



10



10



10



10



10



G3, F & 0.003



Mean



1.94



5.02



8.32



12.40



15.20



18.19



20.54



23.65



±SD



1.17



1.99



2.94



3.60



4.97



5.30



6.10



6.10



n



10



10



10



10



10



10



10



10



G4, F & 0.01



Mean



2.04



5.07



8.26



12.61



15.77



18.78



21.19



24.44



±SD



0.98



2.05



2.61



4.64



6.02



6.16



5.57



5.44



n



10



10



10



10



10



10



10



10



G5, F & 0.03



Mean



2.03



5.19



8.34



12.51



15.00



18.35



20.86



23.62



±SD



1.21



2.72



3.84



5.27



5.91



5.46



6.80



7.11



n



10



10



10



10



10



10



10



10



M: Male; F: Female; SD: Standard Deviation; n: Number of animals


 



  • TABLE 3 (Contd...). SUMMARY OF PERCENT CHANGE IN BODY WEIGHT WITH RESPECT TO DAY 1 RECORD


Refer Appendix 3








































































































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Percent Change in Body Weight (%) during Days



1 to 36



1 to 43



1 to 50



1 to 57



1 to 64



1 to 71



1 to 78



1 to 85



1 to 89



G1, M & 0



Mean



52.65



62.62



71.78



80.23



87.35



103.80



112.51



121.47



125.37



±SD



7.48



7.99



8.77



11.33



11.28



10.83



10.09



10.45



10.59



n



10



10



10



10



10



10



10



10



10



G2, M & 0.001



Mean



56.00



65.85



75.17



84.88



92.55



109.59



118.07



127.25



131.29



±SD



6.03



8.27



11.68



15.21



15.23



15.49



15.07



17.35



16.63



n



10



10



10



10



10



10



10



10



10



G3, M & 0.003



Mean



51.39



61.76



71.99



79.28



86.97



104.27



113.07



121.19



125.60



±SD



6.63



9.56



12.79



14.58



14.78



13.17



12.84



12.86



12.78



n



10



10



10



10



10



10



10



10



10



G4, M & 0.01



Mean



56.63



66.22



77.72



85.16



93.21



108.95



117.44



125.41



130.88



±SD



6.09



9.46



10.52



11.35



11.67



11.24



11.12



11.40



13.02



n



10



10



10



10



10



10



10



10



10



G5, M & 0.03



Mean



56.07



66.93



79.77



85.20



98.50



116.82



125.71



134.92



138.94



±SD



6.19



8.27



11.81



23.48



12.35



13.43



13.17



13.43



13.46



n



10



10



10



10



10



10



10



10



10



G1, F & 0



Mean



29.22



33.33



36.79



41.10



44.87



50.99



54.50



58.14



61.35



±SD



5.34



4.94



4.78



4.82



5.49



5.22



5.12



5.31



5.14



n



10



10



10



10



10



10



10



10



10



G2, F & 0.001



Mean



29.74



33.55



38.28



41.88



45.57



55.12



56.70



60.67



64.29



±SD



10.31



10.70



9.95



10.06



10.13



11.05



11.27



10.99



10.96



n



10



10



10



10



10



10



10



10



10



G3, F & 0.003



Mean



28.92



32.36



35.76



39.91



44.44



50.61



54.58



58.39



61.33



±SD



5.83



4.74



4.78



5.30



5.66



5.45



5.25



5.11



5.27



n



10



10



10



10



10



10



10



10



10



G4, F & 0.01



Mean



28.54



32.69



37.35



40.89



44.95



52.58



56.18



60.17



63.21



±SD



5.79



5.92



5.53



5.34



4.97



4.90



4.86



5.68



5.42



n



10



10



10



10



10



10



10



10



10



G5, F & 0.03



Mean



28.09



31.63



35.05



38.83



42.59



49.69



53.37



56.99



60.45



±SD



6.99



8.11



8.12



8.34



8.19



8.48



8.07



7.80



7.42



n



10



10



10



10



10



10



10



10



10



M: Male; F: Female; SD: Standard Deviation; n: Number of animals


 



  • TABLE 3 (Contd...). SUMMARY OF PERCENT CHANGE IN BODY WEIGHT WITH RESPECT TO DAY 1 RECORD


Refer Appendix 3








































































































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Percent Change in Body Weight (%) during Days



1 to


5



1 to


8



1 to 12



1 to 15



1 to 19



1 to 22



1 to 26



1 to 29



1 to 36



G1R-S1, M & 0



Mean



3.11



10.85



15.99



22.69



26.37



29.90



36.44



41.66



46.88



±SD



1.04



1.26



1.34



1.63



1.90



2.16



4.30



4.16



6.15



n



5



5



5



5



5



5



5



5



5



G2R-S1, M & 0.001



Mean



3.42



9.64



15.01



21.97



25.28



29.06



34.53



39.34



46.79



±SD



1.04



2.16



2.21



2.98



3.87



4.31



4.63



5.35



5.25



n



5



5



5



5



5



5



5



5



5



G3R-S1, M & 0.003



Mean



2.61



10.82



16.30



23.42



26.71



31.07



37.94



43.77



50.37



±SD



1.41



3.12



3.81



5.10



4.27



4.26



4.19



4.68



6.23



n



5



5



5



5



5



5



5



5



5



G4R-S1, M & 0.01



Mean



2.88



10.58



16.61



23.17



26.94



31.26



37.83



42.66



49.62



±SD



1.40



1.42



2.48



2.13



2.28



2.72



3.58



5.73



6.59



n



5



5



5



5



5



5



5



5



5



G5R-S1, M & 0.03



Mean



2.95



10.95



16.87



23.88



27.22



31.57



38.03



44.01



50.08



±SD



0.94



0.88



0.86



1.46



1.36



1.54



2.73



2.98



3.17



n



5



5



5



5



5



5



5



5



5



G1R-S1, F & 0



Mean



1.73



6.25



10.28



13.28



16.11



18.70



20.99



23.32



33.54



±SD



1.21



0.98



2.35



2.25



4.34



4.34



4.12



4.12



5.31



n



5



5



5



5



5



5



5



5



5



G2R-S1, F & 0.001



Mean



1.97



4.94



9.25



12.31



15.09



17.59



20.03



22.43



30.79



±SD



1.20



1.97



1.56



0.65



1.55



1.71



1.55



2.52



4.03



n



5



5



5



5



5



5



5



5



5



G3R-S1, F & 0.003



Mean



2.16



6.82



10.74



14.32



17.19



19.66



22.36



24.92



36.04



±SD



1.27



0.83



2.17



2.73



2.59



2.26



2.06



2.13



6.54



n



5



5



5



5



5



5



5



5



5



G4R-S1, F & 0.01



Mean



1.88



5.93



9.81



13.15



15.84



18.28



21.25



23.85



33.17



±SD



1.05



2.82



4.26



3.89



4.02



3.50



2.80



2.87



3.37



n



5



5



5



5



5



5



5



5



5



G5R-S1, F & 0.03



Mean



1.69



6.14



10.09



13.38



16.12



18.56



21.16



23.80



34.09



±SD



1.27



2.06



2.20



2.56



2.49



2.95



3.31



3.54



7.17



n



5



5



5



5



5



5



5



5



5



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; R: Recovery; S1: Satellite 1.


 



  • TABLE 3 (Contd...). SUMMARY OF PERCENT CHANGE IN BODY WEIGHT WITH RESPECT TO DAY 1 RECORD


Refer Appendix 3








































































































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Percent Change in Body Weight (%) during Days



1 to 43



1 to 50



1 to 57



1 to 64



1 to 71



1 to 78



1 to 85



1 to 92



1 to 99



G1R-S1, M & 0



Mean



58.46



69.63



78.87



87.47



102.89



112.46



119.99



127.39



137.82



±SD



8.79



9.87



10.67



12.32



12.18



12.77



12.68



13.32



14.67



n



5



5



5



5



5



5



5



5



5



G2R-S1, M & 0.001



Mean



61.83



72.89



81.45



87.98



103.98



112.98



118.31



125.90



137.55



±SD



9.33



15.38



16.38



16.67



17.15



17.21



16.01



16.13



16.96



n



5



5



5



5



5



5



5



5



5



G3R-S1, M & 0.003



Mean



64.34



75.75



82.07



88.28



95.17



105.32



111.11



118.57



127.01



±SD



5.86



8.70



9.23



9.63



12.70



12.55



13.73



14.42



14.87



n



5



5



5



5



5



5



5



5



5



G4R-S1, M & 0.01



Mean



66.50



75.77



84.72



91.65



106.55



115.75



123.15



131.22



139.14



±SD



5.88



10.85



11.39



11.31



12.73



13.65



13.20



13.57



14.33



n



5



5



5



5



5



5



5



5



5



G5R-S1, M & 0.03



Mean



62.28



73.49



81.73



87.42



95.33



104.76



111.08



119.24



128.07



±SD



8.58



14.26



16.38



16.61



17.02



16.48



17.04



17.26



17.40



n



5



5



5



5



5



5



5



5



5



G1R-S1, F & 0



Mean



37.08



41.62



45.22



49.14



56.11



60.18



64.19



68.30



74.63



±SD



7.04



9.36



8.96



9.28



9.93



10.63



10.26



10.55



9.92



n



5



5



5



5



5



5



5



5



5



G2R-S1, F & 0.001



Mean



35.60



39.00



42.68



46.20



52.82



54.32



57.76



62.07



66.92



±SD



5.44



5.24



5.65



5.35



4.34



4.12



4.08



3.89



3.46



n



5



5



5



5



5



5



5



5



5



G3R-S1, F & 0.003



Mean



40.58



43.52



47.31



50.68



56.66



61.17



64.86



68.82



72.98



±SD



9.57



9.41



9.43



10.10



9.59



9.91



10.39



10.26



10.70



n



5



5



5



5



5



5



5



5



5



G4R-S1, F & 0.01



Mean



38.33



41.96



46.30



50.53



56.13



59.91



63.04



67.21



70.79



±SD



3.16



4.65



4.44



4.13



3.80



4.00



4.22



4.11



4.02



n



5



5



5



5



5



5



5



5



5



G5R-S1, F & 0.03



Mean



36.49



40.06



44.92



47.98



54.40



58.15



62.36



65.85



69.27



±SD



7.45



7.29



7.26



7.31



6.37



6.65



6.08



6.20



6.87



n



5



5



5



5



5



5



5



5



5



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; R: Recovery; S1: Satellite 1.


 



  • TABLE 3 (Contd...). SUMMARY OF PERCENT CHANGE IN BODY WEIGHT WITH RESPECT TO DAY 1 RECORD


Refer Appendix 3











































































































































































































































































































Group, Sex & Dose (mg/L)


 

Percent Change in Body Weight (%) during Days



1 to 106



1 to 113



1 to 120



1 to 127



1 to 134



1 to 141



G1R-S1, M & 0



Mean



144.25



150.49



157.52



164.81



171.98



174.63



±SD



13.89



12.39



11.43



12.17



12.36



12.06



n



5



5



5



5



5



5



G2R-S1, M & 0.001



Mean



144.55



152.23



159.73



166.93



174.79



178.59



±SD



17.49



17.65



17.51



17.50



17.63



18.08



n



5



5



5



5



5



5



G3R-S1, M & 0.003



Mean



135.33



143.72



151.54



159.34



166.09



170.17



±SD



15.65



14.93



14.80



15.14



15.31



15.72



n



5



5



5



5



5



5



G4R-S1, M & 0.01



Mean



146.04



153.73



161.80



170.66



177.67



181.71



±SD



13.82



14.62



14.87



16.03



15.83



16.17



n



5



5



5



5



5



5



G5R-S1, M & 0.03



Mean



136.14



144.30



152.32



160.62



167.52



171.47



±SD



19.07



18.85



19.43



19.69



20.17



19.67



n



5



5



5



5



5



5



G1R-S1, F & 0



Mean



79.79



85.05



92.43



96.69



99.04



103.79



±SD



9.91



10.29



10.09



10.22



10.30



10.88



n



5



5



5



5



5



5



G2R-S1, F & 0.001



Mean



70.76



77.51



84.53



88.71



91.49



95.97



±SD



3.49



3.23



3.31



3.60



3.58



3.64



n



5



5



5



5



5



5



G3R-S1, F & 0.003



Mean



77.48



82.70



89.02



93.47



96.34



101.15



±SD



10.49



10.31



10.61



10.56



10.57



10.82



n



5



5



5



5



5



5



G4R-S1, F & 0.01



Mean



75.80



81.49



88.68



92.92



95.63



100.44



±SD



3.72



3.20



3.13



3.53



3.75



3.71



n



5



5



5



5



5



5



G5R-S1, F & 0.03



Mean



73.49



79.80



86.37



90.18



93.30



97.67



±SD



6.23



6.20



6.13



5.88



6.52



6.61



n



5



5



5



5



5



5



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; R: Recovery; S1: Satellite 1.


 


TABLE 3 (Contd...). SUMMARY OF PERCENT CHANGE IN BODY WEIGHT WITH RESPECT TO DAY 1 RECORD


Refer Appendix 3











































































































































































































































































































Group, Sex & Dose (mg/L)


 

Percent Change in Body Weight (%) during Days



1 to 148



1 to 155



1 to 162



1 to 169



1 to 176



1 to 179



G1R-S1, M & 0



Mean



182.30



188.57



196.47



203.81



211.17



217.07



±SD



12.73



12.14



12.21



12.92



13.55



13.97



n



5



5



5



5



5



5



G2R-S1, M & 0.001



Mean



185.72



193.23



200.77



208.18



215.30



220.97



±SD



18.64



18.94



19.37



19.87



19.87



18.77



n



5



5



5



5



5



5



G3R-S1, M & 0.003



Mean



177.86



185.22



193.56



201.45



208.81



213.18



±SD



15.92



16.57



17.27



17.17



17.37



16.93



n



5



5



5



5



5



5



G4R-S1, M & 0.01



Mean



188.66



196.15



203.74



211.57



218.98



223.27



±SD



16.17



16.56



16.66



17.08



17.55



16.76



n



5



5



5



5



5



5



G5R-S1, M & 0.03



Mean



179.30



187.15



194.53



201.93



209.94



215.48



±SD



19.65



19.60



19.45



19.69



19.52



18.87



n



5



5



5



5



5



5



G1R-S1, F & 0



Mean



109.60



115.79



120.30



126.19



143.45



146.85



±SD



10.95



10.63



10.49



12.09



28.41



27.84



n



5



5



5



5



5



5



G2R-S1, F & 0.001



Mean



103.09



109.61



115.36



121.35



126.68



129.73



±SD



4.06



4.82



4.86



4.35



4.80



5.29



n



5



5



5



5



5



5



G3R-S1, F & 0.003



Mean



107.45



113.12



118.56



124.38



129.49



133.28



±SD



10.56



10.94



11.56



10.73



11.32



10.91



n



5



5



5



5



5



5



G4R-S1, F & 0.01



Mean



107.37



114.94



120.95



126.61



132.05



135.21



±SD



3.43



3.50



3.63



3.42



2.90



2.68



n



5



5



5



5



5



5



G5R-S1, F & 0.03



Mean



103.75



109.79



116.00



119.70



125.13



128.35



±SD



5.89



5.39



5.38



9.65



9.43



9.55



n



5



5



5



5



5



5



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; R: Recovery; S1: Satellite 1.


 



  • TABLE 3 (Contd...). SUMMARY OF PERCENT CHANGE IN BODY WEIGHT WITH RESPECT TO DAY 1 RECORD


Refer Appendix 3








































































































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Percent Change in Body Weight (%) during Days



1 to


5



1 to


8



1 to 12



1 to 15



1 to 19



1 to 22



1 to 26



1 to 29



1 to 36



G1R-S2, M & 0



Mean



3.04



12.24



17.77



24.57



27.88



31.96



38.04



43.91



51.34



±SD



0.94



1.06



1.63



1.64



1.24



1.99



3.67



3.83



3.74



n



5



5



5



5



5



5



5



5



5



G2R-S2, M & 0.001



Mean



4.76



12.84



18.51



26.90



30.62



34.51



41.12



45.82



52.13



±SD



1.94



1.64



2.39



3.55



4.50



5.45



6.14



6.04



6.79



n



5



5



5



5



5



5



5



5



5



G3R-S2, M & 0.003



Mean



3.03



11.52



17.54



24.85



28.30



32.45



38.74



44.27



50.40



±SD



1.72



2.42



1.98



2.87



2.85



3.91



4.01



4.37



4.17



n



5



5



5



5



5



5



5



5



5



G4R-S2, M & 0.01



Mean



3.89



12.04



17.43



25.61



28.08



32.06



38.29



43.87



49.81



±SD



1.28



1.67



1.60



2.46



2.88



2.13



2.74



3.08



4.30



n



5



5



5



5



5



5



5



5



5



G5R-S2, M & 0.03



Mean



3.40



11.80



18.04



24.79



28.24



32.41



39.08



44.91



50.05



±SD



1.42



1.01



1.72



1.35



2.13



2.52



1.35



1.87



2.28



n



5



5



5



5



5



5



5



5



5



G1R-S2, F & 0



Mean



1.82



5.46



9.48



11.92



14.61



17.36



20.32



22.85



30.45



±SD



1.30



2.91



4.22



5.24



6.31



6.22



6.41



6.09



10.43



n



5



5



5



5



5



5



5



5



5



G2R-S2, F & 0.001



Mean



1.89



5.36



9.22



12.28



15.28



17.86



20.68



22.88



32.29



±SD



1.23



2.66



4.86



4.67



4.43



4.38



4.34



4.84



3.88



n



5



5



5



5



5



5



5



5



5



G3R-S2, F & 0.003



Mean



1.68



5.48



10.00



13.47



16.40



20.06



22.88



25.24



36.84



±SD



1.25



2.39



2.14



1.22



1.52



1.85



2.62



1.94



4.53



n



5



5



5



5



5



5



5



5



5



G4R-S2, F & 0.01



Mean



1.97



5.99



10.10



13.09



16.18



18.79



21.73



24.48



32.79



±SD



1.29



1.83



2.38



2.83



2.32



2.16



2.18



2.38



4.13



n



5



5



5



5



5



5



5



5



5



G5R-S2, F & 0.03



Mean



2.17



5.79



10.08



13.18



16.12



18.73



21.76



24.84



33.71



±SD



1.29



3.15



4.45



4.85



4.52



4.30



3.93



3.80



7.37



n



5



5



5



5



5



5



5



5



5



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; R: Recovery; S2: Satellite 2.
TABLE 3 (Contd...). SUMMARY OF PERCENT CHANGE IN BODY WEIGHT WITH RESPECT TO DAY 1 RECORD


Refer Appendix 3








































































































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Percent Change in Body Weight (%) during Days



1 to 43



1 to 50



1 to 57



1 to 64



1 to 71



1 to 78



1 to 85



1 to 92



1 to 99



G1R-S2, M & 0



Mean



68.07



78.75



86.77



93.31



108.46



117.01



124.56



132.34



141.43



±SD



5.59



8.04



10.32



10.54



10.93



10.74



10.71



10.68



11.04



n



5



5



5



5



5



5



5



5



5



G2R-S2, M & 0.001



Mean



67.57



79.96



86.24



94.84



112.93



120.97



128.52



136.64



146.73



±SD



2.89



5.68



6.11



7.05



5.90



5.92



5.12



5.51



6.06



n



5



5



5



5



5



5



5



5



5



G3R-S2, M & 0.003



Mean



66.29



80.27



90.80



97.07



103.28



110.76



117.74



125.89



135.13



±SD



4.80



9.48



10.21



10.16



12.88



12.69



13.79



13.19



13.81



n



5



5



5



5



5



5



5



5



5



G4R-S2, M & 0.01



Mean



67.36



81.13



90.89



98.57



115.96



123.47



130.25



138.06



146.91



±SD



6.20



10.46



13.20



14.14



12.57



12.39



13.26



13.27



13.14



n



5



5



5



5



5



5



5



5



5



G5R-S2, M & 0.03



Mean



67.76



80.86



88.96



95.26



103.61



109.97



116.35



124.45



134.81



±SD



6.04



10.12



11.34



11.27



13.90



13.49



13.92



14.32



14.70



n



5



5



5



5



5



5



5



5



5



G1R-S2, F & 0



Mean



34.00



37.96



41.34



45.17



52.41



56.17



59.62



64.27



67.84



±SD



11.70



12.17



11.35



11.14



11.15



11.02



11.03



11.09



10.85



n



5



5



5



5



5



5



5



5



5



G2R-S2, F & 0.001



Mean



38.16



41.75



44.59



48.91



54.73



57.64



61.12



64.82



69.15



±SD



5.76



6.57



6.63



6.02



6.46



6.41



6.66



6.65



6.21



n



5



5



5



5



5



5



5



5



5



G3R-S2, F & 0.003



Mean



41.27



44.55



48.80



52.31



58.84



63.19



66.98



70.43



74.04



±SD



4.71



4.88



5.51



5.74



5.78



6.06



6.03



6.07



6.27



n



5



5



5



5



5



5



5



5



5



G4R-S2, F & 0.01



Mean



38.30



44.03



48.14



51.60



58.23



62.21



65.53



69.46



73.02



±SD



6.93



9.06



8.27



8.07



8.00



8.19



8.39



8.51



8.75



n



5



5



5



5



5



5



5



5



5



G5R-S2, F & 0.03



Mean



38.41



41.27



45.55



48.71



54.77



58.30



62.43



66.91



71.85



±SD



10.02



10.08



10.66



10.62



10.52



10.47



10.54



9.63



9.26



n



5



5



5



5



5



5



5



5



5



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; R: Recovery; S2: Satellite 2.


 


TABLE 3 (Contd...). SUMMARY OF PERCENT CHANGE IN BODY WEIGHT WITH RESPECT TO DAY 1 RECORD


Refer Appendix 3








































































































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Percent Change in Body Weight (%) during Days



1 to 106



1 to 113



1 to 120



1 to 127



1 to 134



1 to 141



1 to 148



1 to 155



1 to 162



G1R-S2, M & 0



Mean



149.24



159.34



167.74



175.94



182.93



185.88



193.68



201.52



209.64



±SD



11.49



12.91



13.03



13.52



13.84



14.02



13.52



13.73



13.86



n



5



5



5



5



5



5



5



5



5



G2R-S2, M & 0.001



Mean



155.24



165.07



173.19



180.75



188.11



192.01



200.38



207.83



215.86



±SD



5.48



4.62



4.90



5.55



6.25



5.85



8.13



8.23



9.01



n



5



5



5



5



5



5



5



5



5



G3R-S2, M & 0.003



Mean



143.07



153.05



161.81



170.31



177.22



180.68



188.33



196.10



203.84



±SD



13.48



13.54



12.50



13.20



13.68



13.52



13.54



13.39



13.91



n



5



5



5



5



5



5



5



5



5



G4R-S2, M & 0.01



Mean



156.48



164.72



173.04



180.73



187.94



191.83



199.70



208.04



215.39



±SD



13.40



12.53



11.79



12.02



11.87



11.60



10.94



11.84



11.88



n



5



5



5



5



5



5



5



5



5



G5R-S2, M & 0.03



Mean



143.72



153.05



160.99



168.74



175.41



179.55



186.65



194.76



202.19



±SD



14.02



13.86



12.29



12.53



12.39



12.64



12.77



12.19



12.30



n



5



5



5



5



5



5



5



5



5



G1R-S2, F & 0



Mean



72.48



78.35



85.82



90.13



92.85



97.21



104.24



110.95



117.15



±SD



10.54



10.97



11.20



10.87



10.63



10.52



10.60



10.79



10.90



n



5



5



5



5



5



5



5



5



5



G2R-S2, F & 0.001



Mean



72.92



78.59



85.32



89.76



92.39



96.60



103.46



110.56



108.05



±SD



6.00



5.49



5.07



5.21



5.45



5.20



4.79



5.28



20.49



n



5



5



5



5



5



5



5



5



5



G3R-S2, F & 0.003



Mean



78.04



84.02



90.40



94.50



97.14



101.61



107.70



114.65



120.82



±SD



6.50



6.54



7.08



7.49



7.33



7.71



7.94



8.45



8.89



n



5



5



5



5



5



5



5



5



5



G4R-S2, F & 0.01



Mean



77.04



82.94



90.04



94.01



96.87



100.93



108.45



114.41



118.88



±SD



8.24



7.96



8.33



8.36



8.33



7.93



8.03



8.15



8.38



n



5



5



5



5



5



5



5



5



5



G5R-S2, F & 0.03



Mean



76.10



81.91



89.28



92.88



95.80



99.46



106.97



114.28



118.75



±SD



9.20



8.01



7.79



7.58



7.69



7.86



7.66



7.55



7.49



n



5



5



5



5



5



5



5



5



5



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; R: Recovery; S2: Satellite 2.


 


TABLE 3 (Contd...). SUMMARY OF PERCENT CHANGE IN BODY WEIGHT WITH RESPECT TO DAY 1 RECORD


Refer Appendix 3








































































































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Percent Change in Body Weight (%) during Days



1 to 169



1 to 176



1 to 183



1 to 90



1 to 97



1 to 204



1 to 211



1 to 218



1 to 225



G1R-S2, M & 0



Mean



217.84



225.58



233.56



240.86



248.23



255.33



262.52



269.36



274.27



±SD



13.51



13.09



12.82



12.33



12.61



13.02



13.15



13.84



13.86



n



5



5



5



5



5



5



5



5



5



G2R-S2, M & 0.001



Mean



223.45



232.13



240.16



247.08



253.72



261.29



266.76



273.07



278.48



±SD



9.59



10.01



10.27



10.61



11.04



10.94



11.20



11.45



12.33



n



5



5



5



5



5



5



5



5



5



G3R-S2, M & 0.003



Mean



210.84



218.25



225.83



232.90



239.37



246.91



252.65



259.52



261.86



±SD



14.01



14.31



14.62



14.31



14.46



14.78



16.32



16.17



16.53



n



5



5



5



5



5



5



5



5



5



G4R-S2, M & 0.01



Mean



222.85



230.87



238.73



246.10



252.81



260.08



267.45



273.91



278.66



±SD



11.95



11.83



12.83



12.51



13.04



13.00



13.75



13.33



13.88



n



5



5



5



5



5



5



5



5



5



G5R-S2, M & 0.03



Mean



209.81



218.38



226.96



233.39



240.12



246.12



253.32



259.95



264.44



±SD



12.42



12.06



13.02



12.19



12.36



12.04



11.81



12.30



12.43



n



5



5



5



5



5



5



5



5



5



G1R-S2, F & 0



Mean



122.05



127.08



132.02



137.67



142.06



146.42



150.71



154.49



156.65



±SD



10.65



10.11



10.40



10.62



10.55



10.45



10.65



11.07



11.16



n



5



5



5



5



5



5



5



5



5



G2R-S2, F & 0.001



Mean



121.87



127.50



132.17



136.96



141.24



145.98



150.50



153.96



156.37



±SD



5.71



5.35



5.71



5.26



5.39



4.95



4.95



5.39



5.50



n



5



5



5



5



5



5



5



5



5



G3R-S2, F & 0.003



Mean



125.66



130.33



134.92



140.07



144.32



148.73



153.05



156.71



158.56



±SD



9.47



9.45



9.59



10.01



9.97



10.37



10.24



10.60



10.89



n



5



5



5



5



5



5



5



5



5



G4R-S2, F & 0.01



Mean



123.67



128.53



133.49



138.45



142.00



146.72



151.24



154.70



156.83



±SD



8.25



7.66



7.89



8.46



8.17



8.21



8.22



8.27



8.25



n



5



5



5



5



5



5



5



5



5



G5R-S2, F & 0.03



Mean



123.35



128.77



133.63



138.58



142.40



146.52



151.22



155.20



157.33



±SD



7.61



7.52



7.70



7.87



7.59



7.64



7.28



7.35



7.40



n



5



5



5



5



5



5



5



5



5



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; R: Recovery; S2: Satellite 2.


 


TABLE 3 (Contd...). SUMMARY OF PERCENT CHANGE IN BODY WEIGHT WITH RESPECT TO DAY 1 RECORD


Refer Appendix 3










































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Percent Change in Body Weight (%) during Days



1 to 232



1 to 239



1 to 246



1 to 253



1 to 260



1 to 267



1 to 269



G1R-S2, M & 0



Mean



277.77



280.97



285.49



290.47



293.91



297.43



299.41



±SD



14.22



14.45



16.69



16.42



16.14



16.51



16.84



n



5



5



5



5



5



5



5



G2R-S2, M & 0.001



Mean



281.98



283.99



288.32



294.54



298.86



301.98



304.02



±SD



12.23



12.49



13.17



13.08



12.80



12.71



12.69



n



5



5



5



5



5



5



5



G3R-S2, M & 0.003



Mean



265.08



267.71



275.93



280.58



284.81



288.09



290.05



±SD



16.14



15.84



16.55



16.71



16.43



16.48



16.20



n



5



5



5



5



5



5



5



G4R-S2, M & 0.01



Mean



281.57



284.22



290.55



296.09



300.41



303.90



305.91



±SD



13.83



13.40



14.07



13.88



13.91



14.41



14.75



n



5



5



5



5



5



5



5



G5R-S2, M & 0.03



Mean



267.46



269.45



277.75



281.55



286.07



289.08



290.75



±SD



11.98



11.90



10.65



10.61



10.51



10.75



10.81



n



5



5



5



5



5



5



5



G1R-S2, F & 0



Mean



158.55



163.97



166.66



169.35



171.89



174.91



176.25



±SD



10.81



10.31



10.14



9.95



10.06



10.01



10.19



n



5



5



5



5



5



5



5



G2R-S2, F & 0.001



Mean



159.15



164.62



167.81



170.62



172.68



175.92



177.73



±SD



5.52



5.93



6.12



5.87



5.57



5.36



5.24



n



5



5



5



5



5



5



5



G3R-S2, F & 0.003



Mean



161.63



170.66



173.44



175.82



177.78



180.66



182.11



±SD



11.08



13.48



12.84



13.03



12.97



12.59



12.53



n



5



5



5



5



5



5



5



G4R-S2, F & 0.01



Mean



159.10



169.88



173.59



176.34



178.67



181.26



182.65



±SD



8.31



9.52



9.86



9.72



9.96



9.95



10.02



n



5



5



5



5



5



5



5



G5R-S2, F & 0.03



Mean



159.93



168.24



170.68



174.07



176.35



179.17



180.65



±SD



7.21



8.59



8.54



8.20



8.19



8.10



8.19



n



5



5



5



5



5



5



5



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; R: Recovery; S2: Satellite 2.



  • SUMMARY OF AVERAGE FEED CONSUMPTION (g/rat/day) RECORD


  Refer Appendix 4

































































































































































































































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Week 1



Week 2



Week 3



Week 4



Week 5



Week 6



Week 7



Week 8



Week 9



Week 10



Week 11



Week 12



Week 13



G1, M & 0



Mean



20.56



20.47



20.31



20.33



22.09



22.19



24.74



24.63



24.66



24.43



24.26



24.49



27.25



±SD



0.28



0.06



0.15



0.04



0.20



0.10



0.49



0.43



0.26



0.25



0.32



0.16



0.82



n



5



5



5



5



5



5



5



5



5



5



5



5



5



G2, M & 0.001



Mean



20.51



20.48



20.14



20.16



22.10



22.17



25.00



25.01



25.01



24.84



24.62



24.54



27.25



±SD



0.11



0.07



0.10



0.11



0.17



0.18



0.33



0.15



0.11



0.25



0.30



0.24



1.06



n



5



5



5



5



5



5



5



5



5



5



5



5



5



G3, M & 0.003



Mean



20.60



20.44



20.36



20.25



22.20



22.26



24.89



24.73



24.60



24.42



24.40



24.78



27.02



±SD



0.15



0.15



0.15



0.19



0.25



0.20



0.29



0.25



0.26



0.27



0.15



0.22



0.98



n



5



5



5



5



5



5



5



5



5



5



5



5



5



G4, M & 0.01



Mean



20.39



20.36



20.17



20.25



22.24



22.35



25.07



24.71



24.83



24.61



24.48



24.62



27.04



±SD



0.13



0.08



0.11



0.18



0.10



0.17



0.27



0.23



0.19



0.39



0.29



0.31



0.71



n



5



5



5



5



5



5



5



5



5



5



5



5



5



G5, M & 0.03



Mean



20.64



20.51



20.33



20.28



22.22



22.41



25.02



24.67



24.56



24.34



24.12



24.25



27.47



±SD



0.18



0.06



0.14



0.18



0.23



0.23



0.32



0.46



0.21



0.18



0.21



0.24



0.37



n



5



5



5



5



5



5



5



5



5



5



5



5



5



G1, F & 0



Mean



15.84



15.64



15.59



15.93



15.81



17.02



17.03



17.02



17.23



17.09



17.20



17.14



26.99



±SD



0.15



0.15



0.18



0.28



0.12



0.24



0.11



0.13



0.28



0.20



0.29



0.16



0.44



n



5



5



5



5



5



5



5



5



5



5



5



5



5



G2, F & 0.001



Mean



15.80



15.65



15.79



15.83



15.82



17.02



17.18



17.00



17.03



17.27



17.26



17.14



27.66



±SD



0.08



0.23



0.44



0.25



0.09



0.27



0.29



0.11



0.17



0.16



0.26



0.26



0.42



n



5



5



5



5



5



5



5



5



5



5



5



5



5



G3, F & 0.003



Mean



15.84



15.72



15.88



15.93



15.86



16.95



17.03



16.86



17.07



16.77



16.94



17.00



27.02



±SD



0.17



0.23



0.23



0.30



0.18



0.42



0.26



0.31



0.43



0.31



0.23



0.27



0.46



n



5



5



5



5



5



5



5



5



5



5



5



5



5



G4, F & 0.01



Mean



15.84



15.73



15.69



15.80



15.73



17.04



17.10



16.97



16.96



16.53*



16.99



16.87



26.93



±SD



0.17



0.23



0.30



0.31



0.17



0.34



0.30



0.17



0.33



0.24



0.24



0.26



0.75



n



5



5



5



5



5



5



5



5



5



5



5



5



5



G5, F & 0.03



Mean



15.87



15.69



15.59



15.75



15.95



17.11



17.17



17.06



16.92



16.81



17.08



17.13



27.47



±SD



0.19



0.22



0.15



0.36



0.33



0.32



0.19



0.22



0.38



0.23



0.40



0.44



1.09



n



5



5



5



5



5



5



5



5



5



5



5



5



5



 M: Male; F: Female; SD: Standard Deviation; n: number of cages.


 



  • TABLE 4 (Contd...). SUMMARY OF AVERAGE FEED CONSUMPTION (g/rat/day) RECORD


  Refer Appendix 4

































































































































































































































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Week 1



Week 2



Week 3



Week 4



Week 5



Week 6



Week 7



Week 8



Week 9



Week 10



Week 11



Week 12



Week 13



G1R-S1, M & 0



Mean



24.27



24.35



24.16



24.20



26.37



26.18



29.30



29.07



28.75



28.46



28.26



28.59



28.28



±SD



7.03



6.84



6.76



6.72



7.06



7.02



7.31



7.08



7.03



6.92



6.64



6.79



6.71



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G2R-S1, M & 0.001



Mean



24.72



24.50



24.25



24.26



26.40



26.29



28.61



28.05



28.94



28.27



28.22



28.58



28.51



±SD



7.13



6.94



6.99



6.85



7.39



7.12



7.14



6.52



6.97



6.74



6.49



6.92



6.80



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G3R-S1, M & 0.003



Mean



24.35



24.12



24.15



24.01



26.15



26.34



28.42



28.62



28.41



28.23



28.34



28.70



28.17



±SD



6.74



6.59



6.86



6.60



6.98



7.10



6.40



7.48



7.18



7.04



7.00



7.31



6.77



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G4R-S1, M & 0.01



Mean



24.41



24.32



24.16



24.08



26.54



26.46



28.21



28.66



28.58



28.69



28.25



28.46



28.63



±SD



7.14



6.89



6.82



6.59



7.47



6.97



6.38



7.47



7.36



7.62



7.40



6.97



7.40



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G5R-S1, M & 0.03



Mean



24.45



24.10



24.12



24.14



26.49



26.70



28.51



28.57



28.59



28.45



28.10



28.20



28.51



±SD



6.74



6.60



6.86



6.79



7.57



7.74



6.37



7.35



7.14



7.67



7.14



6.79



7.49



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G1R-S1, F & 0



Mean



17.20



16.78



16.72



16.26



16.30



17.03



17.45



18.21



17.87



17.92



17.74



18.15



18.03



±SD



1.89



1.68



1.59



0.73



0.94



0.40



0.74



1.54



1.42



0.95



1.18



1.62



1.85



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G2R-S1, F & 0.001



Mean



16.85



16.72



16.89



16.75



16.62



17.37



17.73



17.76



17.95



18.10



18.01



18.37



18.42



±SD



1.82



1.54



1.52



1.76



1.39



1.57



1.85



1.43



1.23



1.60



1.69



2.24



2.04



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G3R-S1, F & 0.003



Mean



16.83



16.62



16.71



16.87



17.05



17.73



17.98



17.96



17.90



17.69



18.00



17.70



17.56



±SD



1.57



1.46



1.67



1.88



1.87



1.81



1.77



1.36



1.29



1.28



1.39



1.10



1.14



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G4R-S1, F & 0.01



Mean



16.87



16.64



16.71



16.33



16.52



17.34



17.57



18.03



17.56



17.47



17.78



17.37



17.34



±SD



1.54



1.52



1.43



1.64



1.75



1.56



1.66



1.14



1.00



0.77



0.62



1.12



1.18



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G5R-S1, F & 0.03



Mean



16.94



16.71



16.71



16.71



16.84



17.60



17.74



18.03



17.96



17.97



18.16



17.73



17.60



±SD



1.84



1.59



1.69



1.31



1.84



0.75



0.77



1.53



1.06



1.45



1.60



1.25



1.12



n



3



3



3



3



3



3



3



3



3



3



3



3



3



 M: Male; F: Female; SD: Standard Deviation; n: number of cages; R: Recovery; S1: Satellite 1.


 



  • TABLE 4 (Contd...). SUMMARY OF AVERAGE FEED CONSUMPTION (g/rat/day) RECORD


  Refer Appendix 4

































































































































































































































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Week 14



Week 15



Week 16



Week 17



Week 18



Week 19



Week 20



Week 21



Week 22



Week 23



Week 24



Week 25



Week 26



G1R-S1, M & 0



Mean



27.77



27.85



27.34



27.14



27.26



27.05



26.10



25.96



26.02



25.74



26.10



25.86



34.78



±SD



6.56



6.15



6.20



6.01



6.11



5.83



5.37



5.16



5.30



4.86



5.22



4.84



1.73



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G2R-S1, M & 0.001



Mean



27.83



28.25



26.87



26.57



26.77



26.68



25.91



25.87



25.85



25.90



26.60



26.56



33.28



±SD



6.50



6.92



6.82



6.30



6.50



6.15



5.09



4.92



4.84



4.43



5.56



4.83



1.75



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G3R-S1, M & 0.003



Mean



27.57



28.43



26.94



26.51



26.66



26.54



26.03



25.77



25.74



25.81



26.06



26.24



33.91



±SD



6.23



7.17



5.91



5.53



5.53



5.95



5.48



5.01



4.81



5.11



5.31



4.68



0.76



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G4R-S1, M & 0.01



Mean



28.05



28.52



26.63



26.42



26.65



26.29



25.92



25.73



25.96



25.83



26.74



26.45



33.64



±SD



6.69



6.65



6.51



5.98



6.26



5.84



5.53



5.12



4.91



5.24



5.21



5.09



0.46



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G5R-S1, M & 0.03



Mean



27.96



28.04



26.66



26.20



26.44



26.42



26.20



26.05



25.91



25.64



26.80



26.29



33.29



±SD



6.85



6.32



6.46



5.86



6.06



6.20



5.51



5.31



5.03



4.29



5.67



4.67



1.67



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G1R-S1, F & 0



Mean



18.39



17.50



16.45



16.50



16.65



16.35



16.55



16.53



16.29



15.85



16.64



17.11



23.22



±SD



2.24



1.82



0.48



0.76



1.11



1.10



1.12



0.91



0.69



0.44



0.86



0.61



1.06



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G2R-S1, F & 0.001



Mean



18.32



17.74



16.82



16.48



16.37



16.24



16.54



16.66



16.83



16.43



17.05



16.79



21.02



±SD



1.65



1.62



0.77



0.59



0.74



0.61



0.77



1.02



1.01



0.82



1.07



0.55



2.96



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G3R-S1, F & 0.003



Mean



17.73



16.33



17.14



17.16



17.22



16.91



16.75



16.50



16.48



16.90



16.86



17.42



22.79



±SD



1.24



0.70



1.39



1.37



1.80



1.58



1.49



1.14



1.20



0.23



1.17



1.75



1.12



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G4R-S1, F & 0.01



Mean



17.74



16.44



16.57



16.45



16.65



16.54



16.54



16.45



16.35



16.53



17.19



17.30



21.91



±SD



1.52



0.26



0.43



0.81



1.22



0.65



1.18



0.95



0.64



0.11



0.33



1.12



1.59



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G5R-S1, F & 0.03



Mean



17.44



16.58



16.86



17.00



17.20



16.41



16.74



16.19



16.49



16.52



17.36



17.75



24.27



±SD



0.85



0.64



0.84



0.93



1.24



0.91



1.01



0.33



1.00



0.58



1.02



1.32



0.95



n



3



3



3



3



3



3



3



3



3



3



3



3



3



 M: Male; F: Female; SD: Standard Deviation; n: number of cages; R: Recovery; S1: Satellite 1.


 


TABLE 4 (Contd...). SUMMARY OF AVERAGE FEED CONSUMPTION (g/rat/day) RECORD


  Refer Appendix 4

































































































































































































































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Week 1



Week 2



Week 3



Week 4



Week 5



Week 6



Week 7



Week 8



Week 9



Week 10



Week 11



Week 12



Week 13



G1R-S2, M & 0



Mean



24.74



24.43



24.33



24.33



26.63



26.62



28.60



28.59



28.47



28.22



28.10



28.61



28.44



±SD



7.42



7.25



7.17



7.13



7.09



7.38



6.65



7.16



6.97



6.60



6.46



7.39



6.94



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G2R-S2, M & 0.001



Mean



24.63



24.47



24.27



24.49



26.43



26.35



27.83



28.74



28.63



28.92



28.62



28.62



28.58



±SD



7.15



6.93



6.84



7.29



7.40



7.18



5.90



7.65



7.40



7.74



7.48



7.52



7.22



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G3R-S2, M & 0.003



Mean



24.70



24.44



24.06



23.97



26.11



26.23



28.10



28.77



28.58



28.78



28.58



28.56



28.38



±SD



6.79



6.64



6.63



6.42



6.80



7.01



6.03



7.52



7.32



7.50



7.47



7.37



7.15



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G4R-S2, M & 0.01



Mean



24.95



24.63



24.41



26.95



26.64



26.45



28.06



28.33



27.97



27.83



27.63



28.86



28.60



±SD



7.53



7.24



7.17



11.46



7.73



7.42



5.50



6.37



6.12



5.70



5.70



7.70



7.59



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G5R-S2, M & 0.03



Mean



24.67



24.68



24.24



24.16



26.15



26.23



28.40



28.96



28.79



28.80



28.55



28.61



28.41



±SD



7.41



7.44



6.66



6.52



7.01



7.21



6.11



7.34



7.19



7.06



6.95



7.48



7.28



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G1R-S2, F & 0



Mean



17.05



16.66



16.70



16.67



17.03



18.32



18.41



18.12



18.07



17.69



17.80



17.48



17.32



±SD



1.75



1.57



1.62



1.42



1.98



1.88



1.71



1.53



1.77



1.26



1.31



1.14



1.08



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G2R-S2, F & 0.001



Mean



16.88



16.64



17.00



16.57



16.75



17.61



17.78



18.15



18.05



18.24



18.11



18.14



18.00



±SD



1.83



1.39



1.48



1.52



1.75



1.64



1.66



1.43



1.16



1.38



0.92



1.51



1.59



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G3R-S2, F & 0.003



Mean



16.71



16.77



16.74



17.12



17.09



18.22



17.90



17.81



17.85



17.90



18.03



17.41



17.17



±SD



1.66



1.44



1.56



2.37



2.15



1.99



1.93



1.50



1.60



1.16



1.64



1.08



1.03



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G4R-S2, F & 0.01



Mean



16.77



16.84



17.11



16.53



16.50



17.54



17.88



18.05



17.87



17.52



18.05



17.61



17.45



±SD



1.55



1.95



1.87



1.26



1.04



1.08



1.29



1.86



1.48



0.88



1.46



1.72



1.63



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G5R-S2, F & 0.03



Mean



16.90



16.67



16.68



16.69



16.60



17.77



17.94



17.89



17.77



18.11



17.80



17.89



17.84



±SD



1.53



1.52



1.66



1.60



1.21



1.43



1.74



1.44



1.12



1.74



1.24



1.13



1.21



n



3



3



3



3



3



3



3



3



3



3



3



3



3



 M: Male; F: Female; SD: Standard Deviation; n: number of cages; R: Recovery; S2: Satellite 2.


 


TABLE 4 (Contd...). SUMMARY OF AVERAGE FEED CONSUMPTION (g/rat/day) RECORD


  Refer Appendix 4

































































































































































































































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Week 14



Week 15



Week 16



Week 17



Week 18



Week 19



Week 20



Week 21



Week 22



Week 23



Week 24



Week 25



Week 26



G1R-S2, M & 0



Mean



27.82



28.12



27.14



27.02



26.73



26.45



26.07



25.98



25.88



25.77



26.03



26.30



26.65



±SD



6.51



6.78



6.47



5.90



5.90



5.87



4.83



4.84



4.97



4.75



5.49



5.56



5.41



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G2R-S2, M & 0.001



Mean



27.86



28.09



26.74



26.47



26.54



26.18



26.15



25.93



25.85



25.68



26.33



25.72



26.49



±SD



6.88



6.26



6.72



6.41



6.52



6.46



5.36



4.94



5.00



4.86



5.21



4.90



5.39



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G3R-S2, M & 0.003



Mean



27.80



28.31



26.20



26.66



26.86



26.91



25.99



25.94



25.95



25.44



26.69



26.24



27.42



±SD



6.60



6.68



4.60



5.49



5.64



6.14



5.06



4.96



5.22



4.60



4.46



4.09



5.85



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G4R-S2, M & 0.01



Mean



28.28



28.31



26.85



26.73



26.49



26.35



25.80



25.85



25.70



25.50



26.36



26.30



26.67



±SD



7.33



7.05



6.71



6.04



5.99



5.76



5.02



5.07



4.70



4.75



5.29



5.21



5.30



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G5R-S2, M & 0.03



Mean



27.93



28.15



27.02



26.88



26.65



26.47



25.77



25.85



25.78



25.56



26.65



26.84



27.05



±SD



6.86



6.42



7.03



5.95



5.50



5.85



5.15



4.85



5.01



4.80



5.33



5.73



5.80



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G1R-S2, F & 0



Mean



17.31



16.64



16.98



17.04



17.02



16.46



16.48



16.45



16.28



17.32



16.49



17.17



17.03



±SD



1.39



1.01



1.08



1.26



1.83



1.06



1.09



0.82



0.85



1.04



0.90



1.86



1.27



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G2R-S2, F & 0.001



Mean



17.84



17.13



16.68



16.85



16.53



16.54



16.40



16.35



16.58



17.11



17.14



17.31



17.50



±SD



1.51



0.92



0.64



1.15



0.92



0.73



0.78



0.66



1.19



1.42



0.74



1.28



1.45



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G3R-S2, F & 0.003



Mean



17.45



16.63



16.49



16.49



16.60



16.41



16.50



16.39



16.01



16.61



16.37



16.52



17.61



±SD



1.13



0.61



0.49



0.78



1.12



0.78



0.95



0.32



0.63



0.90



0.83



0.89



1.79



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G4R-S2, F & 0.01



Mean



17.31



16.65



16.75



16.44



16.45



16.17



16.18



16.36



16.35



17.20



16.56



16.81



16.89



±SD



1.48



0.97



0.88



0.62



0.63



0.52



0.51



0.75



1.23



1.34



0.17



0.26



0.42



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G5R-S2, F & 0.03



Mean



17.75



17.24



16.42



16.48



16.65



16.36



16.41



16.20



16.10



16.97



15.93



16.52



16.85



±SD



1.12



0.88



0.26



0.60



1.07



0.82



0.62



0.49



1.01



1.09



0.29



0.62



0.96



n



3



3



3



3



3



3



3



3



3



3



3



3



3



 M: Male; F: Female; SD: Standard Deviation; n: number of cages; R: Recovery; S2: Satellite 2.


 


TABLE 4 (Contd...). SUMMARY OF AVERAGE FEED CONSUMPTION (g/rat/day) RECORD


  Refer Appendix 4

































































































































































































































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Week 27



Week 28



Week 29



Week 30



Week 31



Week 32



Week 33



Week 34



Week 35



Week 36



Week 37



Week 38



Week 39



G1R-S2, M & 0



Mean



27.13



26.88



27.55



28.29



28.16



27.90



28.56



27.58



25.68



26.71



27.05



27.52



56.68



±SD



5.31



5.65



6.01



6.32



6.55



6.96



7.62



6.60



5.84



6.73



5.64



7.28



20.86



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G2R-S2, M & 0.001



Mean



26.01



26.73



27.70



28.00



28.23



28.01



28.13



27.34



25.10



26.45



26.47



27.27



61.79



±SD



5.03



4.96



5.55



5.52



6.08



6.05



6.10



5.46



5.49



6.02



5.16



6.49



10.39



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G3R-S2, M & 0.003



Mean



26.86



26.74



27.39



28.34



28.55



28.09



28.62



27.69



27.04



26.27



26.62



27.55



61.53



±SD



5.37



5.47



5.73



6.07



6.29



6.57



6.89



6.68



5.87



5.84



5.96



6.66



12.82



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G4R-S2, M & 0.01



Mean



26.21



26.24



27.70



27.28



27.51



28.04



28.34



27.54



25.53



26.45



26.47



27.10



59.96



±SD



5.32



5.08



5.89



4.59



5.13



6.40



6.31



5.81



4.45



5.72



5.75



5.79



13.40



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G5R-S2, M & 0.03



Mean



26.54



26.87



26.95



27.61



27.57



27.85



28.61



27.83



26.83



27.14



27.08



27.10



42.14



±SD



5.29



5.35



5.78



6.70



6.29



7.10



7.18



6.81



4.60



7.23



7.26



5.77



6.27



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G1R-S2, F & 0



Mean



17.69



18.54



18.94



19.07



19.61



18.89



18.14



16.37



16.37



17.55



17.95



17.67



11.42



±SD



1.83



1.36



2.17



2.08



2.69



2.22



2.08



3.04



2.17



1.40



2.94



2.86



1.15



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G2R-S2, F & 0.001



Mean



17.38



18.65



18.55



18.82



19.85



18.89



17.36



16.62



16.97



16.60



17.29



17.09



11.61



±SD



1.03



2.58



1.90



2.36



3.20



2.17



0.84



0.22



1.49



1.01



1.46



1.41



0.73



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G3R-S2, F & 0.003



Mean



17.71



18.91



19.01



19.23



19.74



19.51



18.45



19.44



18.76



17.74



18.13



18.35



10.75



±SD



1.56



1.84



2.25



2.10



3.51



2.22



1.62



2.61



2.29



2.06



1.77



2.35



1.44



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G4R-S2, F & 0.01



Mean



17.09



18.27



18.45



18.72



19.19



19.54



18.43



19.08



18.46



17.38



17.97



18.94



12.24



±SD



0.68



2.19



2.26



2.25



2.54



3.70



2.77



3.37



3.14



1.72



2.01



2.62



1.78



n



3



3



3



3



3



3



3



3



3



3



3



3



3



G5R-S2, F & 0.03



Mean



17.32



18.38



19.21



19.34



19.60



19.42



18.66



19.12



18.78



17.40



17.92



17.81



12.15



±SD



0.87



2.17



2.69



2.93



2.76



3.26



2.62



2.91



2.26



1.13



1.82



1.50



0.83



n



3



3



3



3



3



3



3



3



3



3



3



3



3



 M: Male; F: Female; SD: Standard Deviation; n: number of cages; R: Recovery; S2: Satellite 2.


 


 



  • SUMMARY OF NEUROLOGICAL/FUNCTIONAL OBSERVATION BATTERY (FOB)

    • Refer Appendix 5













































































































































































Week 13



Parameters↓



Group & Sex



G1 & M



G5 & M



G1 & F



G5 & F



Dose (mg/L)



0



0.03



0



0.03



Number of Animals



10



10



10



10



Home cage observations



Home Cage posture



1



1



1



1



Respiratory pattern



1



1



1



1



Clonic involuntary movements



1



1



1



1



Tonic involuntary movements



1



1



1



1



Vocalization



1



1



1



1



Palpebral Closure



1



1



1



1



Handling observation



 



 



Ease of removal from the cage



2



2



2



2



Ease of handling animal in hand



2



2



2



2



Red or crusty deposits



Eyes



1



1



1



1



Nose



1



1



1



1



Mouth



1



1



1



1



Lacrimation



1



1



1



1



Salivation



1



1



1



1



Fur Appearance



1



1



1



1



Piloerection



1



1



1



1



Eye Prominence



1



1



1



1



Muscle Tone



1



1



1



1



Home cage observations: Home cage posture - 1=Normal, Respiratory Pattern - 1=Normal, Clonic involuntary movements - 1=None/Normal, Tonic involuntary movements - 1=None/Normal, Vocalization - 1=Absent (Normal), Palpebral closure - 1=Eyelids wide open (Normal), Handling observation : Ease of removal from the cage - 2=Normal, Ease of handling animal in hand - 2=Normal, Red/Crusty deposits - 1=Absent, Lacrimation - 1=None, Salivation- 1=Absent (Normal), Fur Appearance - 1=Normal, Piloerection - 1=None,  Eye Prominence - 1=Normal, Muscle tone - 1=Normal


       

M: Male; F: Female


 


TABLE 5 (Contd...). SUMMARY OF NEUROLOGICAL/FUNCTIONAL OBSERVATION BATTERY (FOB) RECORD


Refer Appendix 5













































































































































































































Week 13



Parameters↓



Group & Sex



G1 & M



G5 & M



G1 & F



G5 & F



Dose (mg/L)



0



0.03



0



0.03



Number of Animals



10



10



10



10



Open field Observation



Mobility



1



1



1



1



Gait



1



1



1



1



Arousal



3



3



3



3



Number of Rearing



Mean



6.3



6.6



8.6



8.3



±SD



1.1



1.8



1.1



1.3



Numbers of Urination



Mean



1.1



1.1



1.2



1.2



±SD



0.3



0.3



0.4



0.4



Number of Defecation



Mean



1.1



1.1



1.1



1.1



±SD



0.3



0.6



0.6



0.6



Clonic involuntary movement



1



1



1



1



Tonic involuntary movement



1



1



1



1



Stereotype Behaviour



1



1



1



1



Excessive Grooming



Mean



4.8



5.0



4.7



4.6



±SD



0.6



0.7



0.7



0.5



Sensory Observations



 



Approach Response



2



2



2



2



Auditory Response



2



2



2



2



Touch Response



2



2



2



2



Pupil Reflex



2



2



2



2



Tail Pinch Response



2



2



2



2



Righting Reflex



1



1



1



1



Physiological observation


 

 



Body temperature (°F)



Mean



96.9



96.7



98.8



98.7



±SD



0.2



0.3



0.3



0.4



Open field Observation:  Mobility - 1=Normal, Gait - 1=Normal, Arousal - 3=Normal, Clonic involuntary movements - 1=None/Normal, Tonic involuntary movements - 1=None/Normal, Stereotype Behaviour - 1=Absent, Sensory Observations: Approach response - 2=Normal, Auditory Response - 2=Normal, Touch Response - 2=Normal, Pupil Reflex - 2=Normal, Tail Pinch Response - 2=Normal, Righting Reflex - 1=Present



M: Male; F: Female


 


TABLE 5 (Contd...). SUMMARY OF NEUROLOGICAL/FUNCTIONAL OBSERVATION BATTERY (FOB) RECORD


Refer Appendix 5











































































































































































Day 177



Parameters↓



Group & Sex



G1R-S1 & M



G5R-S1 & M



G1R-S1 & F



G5R-S1 & F



Dose (mg/L)



0



0.03



0



0.03



Number of Animals



5



5



5



5



Home cage observations



Home Cage posture



1



1



1



1



Respiratory pattern



1



1



1



1



Clonic involuntary movements



1



1



1



1



Tonic involuntary movements



1



1



1



1



Vocalization



1



1



1



1



Palpebral Closure



1



1



1



1



Handling observation



 



 



Ease of removal from the cage



2



2



2



2



Ease of handling animal in hand



2



2



2



2



Red or crusty deposits



Eyes



1



1



1



1



Nose



1



1



1



1



Mouth



1



1



1



1



Lacrimation



1



1



1



1



Salivation



1



1



1



1



Fur Appearance



1



1



1



1



Piloerection



1



1



1



1



Eye Prominence



1



1



1



1



Muscle Tone



1



1



1



1



Home cage observations: Home cage posture - 1=Normal, Respiratory Pattern - 1=Normal, Clonic involuntary movements - 1=None/Normal, Tonic involuntary movements - 1=None/Normal, Vocalization - 1=Absent (Normal), Palpebral closure - 1=Eyelids wide open (Normal), Handling observation : Ease of removal from the cage - 2=Normal, Ease of handling animal in hand - 2=Normal, Red/Crusty deposits - 1=Absent, Lacrimation - 1=None, Salivation- 1=Absent (Normal), Fur Appearance - 1=Normal, Piloerection - 1=None,  Eye Prominence - 1=Normal, Muscle tone - 1=Normal


       

M: Male; F: Female; R: Recovery; S1: Satellite 1.


 


TABLE 5 (Contd...). SUMMARY OF NEUROLOGICAL/FUNCTIONAL OBSERVATION BATTERY (FOB) RECORD


Refer Appendix 5













































































































































































































Day 177



Parameters↓



Group & Sex



G1R-S1 & M



G5R-S1 & M



G1R-S1 & F



G5R-S1 & F



Dose (mg/L)



0



0.03



0



0.03



Number of Animals



5



5



5



5



Open field Observation



Mobility



1



1



1



1



Gait



1



1



1



1



Arousal



3



3



3



3



Number of Rearing



Mean



6.0



5.4



5.8



6.0



±SD



1.0



1.1



0.8



0.7



Numbers of Urination



Mean



1.2



1.2



1.2



1.2



±SD



0.4



0.4



0.4



0.4



Number of Defecation



Mean



1.0



1.2



1.0



1.0



±SD



0.7



0.8



0.7



0.7



Clonic involuntary movement



1



1



1



1



Tonic involuntary movement



1



1



1



1



Stereotype Behaviour



1



1



1



1



Excessive Grooming



Mean



4.0



4.8



4.0



4.0



±SD



0.7



0.8



0.7



0.7



Sensory Observations



 



Approach Response



2



2



2



2



Auditory Response



2



2



2



2



Touch Response



2



2



2



2



Pupil Reflex



2



2



2



2



Tail Pinch Response



2



2



2



2



Righting Reflex



1



1



1



1



Physiological observation


 

 



Body temperature (°F)



Mean



96.8



96.8



97.2



97.2



±SD



0.3



0.2



0.2



0.2



Open field Observation:  Mobility - 1=Normal, Gait - 1=Normal, Arousal - 3=Normal, Clonic involuntary movements - 1=None/Normal, Tonic involuntary movements - 1=None/Normal, Stereotype Behaviour - 1=Absent, Sensory Observations: Approach response - 2=Normal, Auditory Response - 2=Normal, Touch Response - 2=Normal, Pupil Reflex - 2=Normal, Tail Pinch Response - 2=Normal, Righting Reflex - 1=Present



M: Male; F: Female; R: Recovery; S1: Satellite 1.


 


TABLE 5 (Contd...). SUMMARY OF NEUROLOGICAL/FUNCTIONAL OBSERVATION BATTERY (FOB) RECORD


Refer Appendix 5











































































































































































Day 267



Parameters↓



Group & Sex



G1R-S2 & M



G5R-S2 & M



G1R-S2 & F



G5R-S2 & F



Dose (mg/L)



0



0.03



0



0.03



Number of Animals



5



5



5



5



Home cage observations



Home Cage posture



1



1



1



1



Respiratory pattern



1



1



1



1



Clonic involuntary movements



1



1



1



1



Tonic involuntary movements



1



1



1



1



Vocalization



1



1



1



1



Palpebral Closure



1



1



1



1



Handling observation



 



 



Ease of removal from the cage



2



2



2



2



Ease of handling animal in hand



2



2



2



2



Red or crusty deposits



Eyes



1



1



1



1



Nose



1



1



1



1



Mouth



1



1



1



1



Lacrimation



1



1



1



1



Salivation



1



1



1



1



Fur Appearance



1



1



1



1



Piloerection



1



1



1



1



Eye Prominence



1



1



1



1



Muscle Tone



1



1



1



1



Home cage observations: Home cage posture - 1=Normal, Respiratory Pattern - 1=Normal, Clonic involuntary movements - 1=None/Normal, Tonic involuntary movements - 1=None/Normal, Vocalization - 1=Absent (Normal), Palpebral closure - 1=Eyelids wide open (Normal), Handling observation : Ease of removal from the cage - 2=Normal, Ease of handling animal in hand - 2=Normal, Red/Crusty deposits - 1=Absent, Lacrimation - 1=None, Salivation- 1=Absent (Normal), Fur Appearance - 1=Normal, Piloerection - 1=None,  Eye Prominence - 1=Normal, Muscle tone - 1=Normal


       

M: Male; F: Female; R: Recovery; S2: Satellite 2.


 


TABLE 5 (Contd...). SUMMARY OF NEUROLOGICAL/FUNCTIONAL OBSERVATION BATTERY (FOB) RECORD


Refer Appendix 5













































































































































































































Day 267



Parameters↓



Group & Sex



G1R-S2 & M



G5R-S2 & M



G1R-S2 & F



G5R-S2 & F



Dose (mg/L)



0



0.03



0



0.03



Number of Animals



5



5



5



5



Open field Observation



Mobility



1



1



1



1



Gait



1



1



1



1



Arousal



3



3



3



3



Number of Rearing



Mean



6.4



6.4



7.2



7.2



±SD



0.9



1.3



1.8



2.3



Numbers of Urination



Mean



1.2



1.2



1.4



1.4



±SD



0.4



0.4



0.9



0.5



Number of Defecation



Mean



1.0



1.0



1.2



1.2



±SD



0.7



0.7



0.4



0.8



Clonic involuntary movement



1



1



1



1



Tonic involuntary movement



1



1



1



1



Stereotype Behaviour



1



1



1



1



Excessive Grooming



Mean



4.0



4.0



4.4



4.2



±SD



0.7



1.0



1.1



0.4



Sensory Observations



 



Approach Response



2



2



2



2



Auditory Response



2



2



2



2



Touch Response



2



2



2



2



Pupil Reflex



2



2



2



2



Tail Pinch Response



2



2



2



2



Righting Reflex



1



1



1



1



Physiological observation


 

 



Body temperature (°F)



Mean



97.1



97.1



98.4



98.5



±SD



0.5



0.4



0.4



0.4



Open field Observation:  Mobility - 1=Normal, Gait - 1=Normal, Arousal - 3=Normal, Clonic involuntary movements - 1=None/Normal, Tonic involuntary movements - 1=None/Normal, Stereotype Behaviour - 1=Absent, Sensory Observations: Approach response - 2=Normal, Auditory Response - 2=Normal, Touch Response - 2=Normal, Pupil Reflex - 2=Normal, Tail Pinch Response - 2=Normal, Righting Reflex - 1=Present



M: Male; F: Female; R: Recovery; S2: Satellite 2.


 



  • SUMMARY OF HIND LIMB FOOT SPLAY (cm) RECORD


                                                                                                                                                         Refer Appendix 6










































































































































































# Week 13, Day 177 & 267



Group, Sex & Dose (mg/L)



 



Hind Limb Foot Splay (cm)



G1, M & 0



Mean



7.3



±SD



1.9



n



10



G5, M & 0.03



Mean



6.7



±SD



0.8



n



10



G1, F & 0



Mean



8.4



±SD



2.1



n



10



G5, F & 0.03



Mean



6.8



±SD



1.4



n



10



G1R-S1, M & 0



Mean



7.3



±SD



0.7



n



5



G5R-S1, M & 0.03



Mean



7.1



±SD



0.6



n



5



G1R-S1, F & 0



Mean



6.8



±SD



0.8



n



5



G5R-S1, F & 0.03



Mean



6.8



±SD



0.8



n



5



G1R-S2, M & 0



Mean



5.9



±SD



0.2



n



5



G5R-S2, M & 0.03



Mean



5.9



±SD



0.2



n



5



G1R-S2, F & 0



Mean



6.7



±SD



0.5



n



5



G5R-S2, F & 0.03



Mean



7.0



±SD



0.6



n



5



M: Male; F: Female; R: Recovery; SD: Standard Deviation; n: number of animals; R: Recovery; S1: Satellite 1; S2: Satellite 2. # Week 13 for main group animals, Day 177 & 267 for Recovery Groups


 


 



  • SUMMARY OF ACTIMETER READING RECORD


                                                                                                                                                         Refer Appendix 7










































































































































































# Week 13, Day 177 & 267



Group, Sex & Dose (mg/L)


 

Movement Counts (no.)



G1, M & 0



Mean



1865.1



±SD



32.6



n



10



G5, M & 0.03



Mean



1878.6



±SD



11.9



n



10



G1, F & 0



Mean



2158.6



±SD



30.4



n



10



G5, F & 0.03



Mean



2116.2*



±SD



11.7



n



10



G1R-S1, M & 0



Mean



1879.8



±SD



11.2



n



5



G5R-S1, M & 0.03



Mean



1868.6



±SD



16.8



n



5



G1R-S1, F & 0



Mean



2259.2



±SD



11.3



n



5



G5R-S1, F & 0.03



Mean



2261.6



±SD



10.5



n



5



G1R-S2, M & 0



Mean



1848.0



±SD



12.5



n



5



G5R-S2, M & 0.03



Mean



1839.2



±SD



18.6



n



5



G1R-S2, F & 0



Mean



2264.4



±SD



9.7



n



5



G5R-S2, F & 0.03



Mean



2263.6



±SD



14.0



n



5



M: Male; F: Female; SD: Standard Deviation; n: number of animals; R: Recovery; S1: Satellite 1;  


S2: Satellite 2. # Week 13 for main group animals, Day 177 & 267 for Recovery Groups



  •  

  •  


 



  • SUMMARY OF GRIP STRENGTH (kgf) MEASUREMENT RECORD


Refer Appendix 8
























































































































































































































































# Week 13, Day 177 & 267


 

Group, Sex & Dose (mg/L)


 

Fore limb



Hind limb


 
 

G1, M & 0



Mean



1.486



0.505


 

±SD



0.044



0.051


 

n



10



10


 

G5, M & 0.03



Mean



1.515



0.507


 

±SD



0.033



0.043


 

n



10



10


 

G1, F & 0



Mean



1.370



0.452


 

±SD



0.021



0.024


 

n



10



10


 

G5, F & 0.03



Mean



1.387



0.421


 

±SD



0.038



0.041


 

n



10



10


 

G1R-S1, M & 0



Mean



1.547



0.499


 

±SD



0.017



0.035


 

n



5



5


 

G5R-S1, M & 0.03



Mean



1.525



0.489


 

±SD



0.014



0.036


 

n



5



5


 

G1R-S1, F & 0



Mean



1.432



0.426


 

±SD



0.027



0.049


 

n



5



5


 

G5R-S1, F & 0.03



Mean



1.424



0.421


 

±SD



0.029



0.038


 

n



5



5


 

G1R-S2, M & 0



Mean



1.537



0.478


 

±SD



0.069



0.034


 

n



5



5


 

G5R-S2, M & 0.03



Mean



1.523



0.469


 

±SD



0.023



0.023


 

n



5



5


 

G1R-S2, F & 0



Mean



1.362



0.421


 

±SD



0.045



0.064


 

n



5



5


 

G5R-S2, F & 0.03



Mean



1.420



0.417


 

±SD



0.034



0.036


 

n



5



5


 

M: Male; F: Female; SD: Standard Deviation; n: number of animals; R: Recovery; S1: Satellite 1;  


S2: Satellite 2. # Week 13 for main group animals, Day 177 & 267 for Recovery Groups.


 


 


 


 


 


 


 


 



  • SUMMARY OF OPHTHALMOSCOPIC EXAMINATION RECORD


                                                                                                                                                                                                                                                             Refer Appendix 9


































































































































































































































































Week 13, 26 and 39



Group, Sex & Dose


(mg/L)



G1, M & 0



G5, M & 0.03



G1, F & 0



G5, F & 0.03



G1R-S1, M & 0



G5R-S1, M & 0.03



G1R-S1, F & 0



G5R-S1, F & 0.03



G1R-S2, M & 0



G5R-S2, M & 0.03



G1R-S2, F & 0



G5R-S2, F & 0.03



Number of Animals



10



10



10



10



5



5



5



5



5



5



5



5



Observations


Eye Parameters ↓



Eye



L



R



L



R



L



R



L



R



L



R



L



R



L



R



L



R



L



R



L



R



L



R



L



R



Eye Lids



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



Cornea



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



Iris



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



Aqueous Humour



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



Lens



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



Vitreous Humour



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



Retina/Optic disc



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N



N: No Abnormality Detected; L: Left; R: Right; M: Male; F: Female; R: Recovery; S1: Satellite 1; S2: Satellite 2.


 



  • SUMMARY OF HAEMATOLOGY RECORD


                                          Refer Appendix 10

























































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Total


Leucocyte Count



Total


Erythrocyte Count



Hemoglobin



Haematocrit



Mean Corpuscular Volume



Mean Corpuscular Hemoglobin



Mean Corpuscular Hemoglobin Concentration



(WBC)



(RBC)



(HGB)



(HCT)



(MCV)



(MCH)



(MCHC)



(103 cells/µL)



(106 cells/µL)



(g/dL)



(%)



(fL)



(pg)



(g/dL)



G1, M & 0



Mean



8.41



8.39



16.78



48.53



57.93



20.06



34.59



±SD



1.07



0.45



0.52



1.97



2.45



1.38



1.55



n



10



10



10



10



10



10



10



G2, M & 0.001



Mean



10.19*



8.93



16.60



48.62



54.77



18.60*



34.04



±SD



1.91



1.07



1.89



4.34



3.56



0.68



1.37



n



10



10



10



10



10



10



10



G3, M & 0.003



Mean



9.26



9.33*



16.89



50.04



54.74



18.29*



33.69



±SD



0.89



1.32



1.42



1.97



8.94



1.58



2.10



n



10



10



10



10



10



10



10



G4, M & 0.01



Mean



9.01



9.43*



16.89



49.70



52.71*



17.91*



34.00



±SD



1.23



0.36



0.53



1.24



1.56



0.49



0.46



n



10



10



10



10



10



10



10



G5, M & 0.03



Mean



9.29



9.47*



16.94



50.07



52.92



17.90*



33.83



±SD



1.96



0.28



0.47



1.65



1.81



0.49



0.55



n



10



10



10



10



10



10



10



G1, F & 0



Mean



9.67



8.56



16.94



46.28



54.17



19.93



36.73



±SD



1.77



0.64



0.56



3.03



1.78



1.79



2.48



n



10



10



10



10



10



10



10



G2, F & 0.001



Mean



8.41



8.58



16.34



46.07



53.46



19.01



35.59



±SD



1.16



0.34



0.40



0.98



1.86



0.55



0.50



n



10



10



10



9



10



10



10



G3, F & 0.003



Mean



7.37*



8.56



16.27



46.33



54.15



19.03



35.14*



±SD



1.45



0.34



0.46



1.12



1.63



0.87



0.62



n



10



10



10



10



10



10



10



G4, F & 0.01



Mean



7.78*



7.83



15.58*



44.98



58.40*



18.11



34.56*



±SD



1.41



1.27



1.75



3.39



6.89



6.54



1.54



n



10



10



10



10



10



10



10



G5, F & 0.03



Mean



7.68*



8.70



16.55



47.33



54.38



19.02



34.95*



±SD



1.85



0.36



0.64



1.80



1.45



0.43



0.59



n



10



10



10



10



10



10



10



M: Male; F: Female; SD: Standard Deviation; n: number of animals; *: Statistically significant (p<0.05).


TABLE 10 (Contd…).  SUMMARY OF HAEMATOLOGY RECORD


                                                                                       Refer Appendix 10



























































































































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Platelet Count



Mean Platelet Volume



Reticulocyte Count



Neutrophils



Lymphocytes



Monocytes



Eosinophils



Basophils



Absolute Reticulocyte Count



(PLT)



(MPV)



(Retic)



(Neut)



(Lymph)



(Mono)



(Eos)



(Baso)



(Retic)



(103 cells/µL)



(fL)



(%)



(%)



(%)



(%)



(%)



(%)



(109 cells/L)



G1, M & 0



Mean



992.00



6.13



3.96



22.50



71.11



3.14



1.83



0.26



329.92



±SD



144.45



0.39



1.07



3.27



5.01



0.90



1.59



0.12



80.64



n



10



10



10



10



10



10



10



10



10



G2, M & 0.001



Mean



916.20



6.16



3.13



28.48



66.10



2.97



1.39



0.20



251.70



±SD



135.95



0.38



3.00



3.15



3.36



0.72



0.45



0.07



158.55



n



10



10



10



10



10



10



10



10



10



G3, M & 0.003



Mean



992.50



6.23



4.29



30.76*



64.34*



2.69



1.20



0.19



305.20



±SD



192.54



0.71



8.44



6.06



6.57



0.72



0.56



0.06



470.60



n



10



10



10



10



10



10



10



10



10



G4, M & 0.01



Mean



991.60



5.97



2.26



31.77*



63.59*



2.57



1.27



0.16*



212.41



±SD



126.11



0.26



0.44



7.91



8.04



0.87



0.22



0.05



40.31



n



10



10



10



10



10



10



10



10



10



G5, M & 0.03



Mean



939.00



6.19



1.86



28.98*



66.06



2.54



1.58



0.21



176.10



±SD



224.60



0.28



0.34



5.33



5.14



0.81



0.49



0.09



32.10



n



10



10



10



10



10



10



10



10



10



G1, F & 0



Mean



939.60



5.63



2.35



26.54



68.11



2.79



1.30



0.22



199.91



±SD



238.96



0.29



0.53



6.16



6.16



0.67



0.43



0.11



42.79



n



10



10



10



10



10



10



10



10



10



G2, F & 0.001



Mean



980.80



5.71



1.68



25.34



68.66



3.46



1.41



0.20



143.95



±SD



190.68



0.15



0.34



3.17



3.55



0.85



0.44



0.05



27.61



n



10



10



10



10



10



10



10



10



10



G3, F & 0.003



Mean



874.30



5.92



1.54



26.92



68.14



2.43



1.62



0.21



131.63



±SD



186.83



0.25



0.29



5.90



6.13



0.64



0.53



0.07



21.51



n



10



10



10



10



10



10



10



10



10



G4, F & 0.01



Mean



945.20



6.71



4.94



27.77



67.14



2.50



1.53



0.23



311.88



±SD



309.29



2.30



7.15



6.06



6.33



0.77



1.16



0.21



368.27



n



10



10



10



10



10



10



10



10



10



G5, F & 0.03



Mean



959.00



5.92



1.51



28.79



65.11



2.71



2.34



0.23



131.38



±SD



275.55



0.53



0.21



6.17



7.84



0.70



1.81



0.11



17.65



n



10



10



10



10



10



10



10



10



10



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; *: Statistically significant (p<0.05)


 


TABLE 10 (Contd…). SUMMARY OF HAEMATOLOGY RECORD


    Refer Appendix 10

























































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Absolute Neutrophils



Absolute Lymphocytes



Absolute Monocytes



Absolute Eosinophils



Absolute Basophils



Prothrombin Time



Activated Prothrombin Time



(Neut)



(Lymph)



(Mono)



(Eos)



(Baso)



(PT)



(APTT)



(103 cells/µL)



(103 cells/µL)



(103 cells/µL)



(103 cells/µL)



(103 cells/µL)



(Seconds)



(Seconds)



G1, M & 0



Mean



1.91



5.95



0.27



0.16



0.02



12.19



21.50



±SD



0.46



0.70



0.08



0.15



0.01



1.37



2.28



n



10



10



10



10



10



10



10



G2, M & 0.001



Mean



2.92*



6.71



0.31



0.14



0.02



14.71*



20.76



±SD



0.70



1.20



0.12



0.04



0.01



0.66



4.00



n



10



10



10



10



10



10



10



G3, M & 0.003



Mean



2.85*



5.96



0.25



0.11



0.02



14.66*



23.16



±SD



0.62



0.85



0.07



0.06



0.00



1.04



3.93



n



10



10



10



10



10



10



10



G4, M & 0.01



Mean



2.85*



5.75



0.23



0.11



0.01



14.85*



22.48



±SD



0.74



1.09



0.08



0.03



0.01



0.45



4.75



n



10



10



10



10



10



10



10



G5, M & 0.03



Mean



2.62*



6.21



0.24



0.15



0.02



14.58*



24.08



±SD



0.37



1.70



0.09



0.05



0.01



1.02



1.97



n



10



10



10



10



10



10



10



G1, F & 0



Mean



2.48



6.68



0.27



0.13



0.02



16.78



20.98



±SD



0.25



1.77



0.08



0.05



0.01



1.14



4.05



n



10



10



10



10



10



10



10



G2, F & 0.001



Mean



2.13



5.79



0.29



0.12



0.02



15.84



23.24



±SD



0.39



0.92



0.06



0.03



0.01



0.82



4.31



n



10



10



10



10



10



10



10



G3, F & 0.003



Mean



2.00



5.01*



0.18*



0.12



0.02



16.83



23.07



±SD



0.62



1.02



0.07



0.05



0.01



2.03



3.00



n



10



10



10



10



10



10



10



G4, F & 0.01



Mean



2.19



5.20*



0.20



0.12



0.02



16.97



26.84*



±SD



0.76



0.87



0.08



0.10



0.02



1.25



6.04



n



10



10



10



10



10



10



10



G5, F & 0.03



Mean



2.15



5.07*



0.21



0.17



0.02



16.02



23.40



±SD



0.50



1.58



0.07



0.11



0.01



0.73



3.02



n



10



10



10



10



10



10



10



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; *: Statistically significant (p<0.05).


 


 


TABLE 10 (Contd…).  SUMMARY OF HAEMATOLOGY RECORD


                                                                                       Refer Appendix 10

























































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Total


Leucocyte Count



Total


Erythrocyte Count



Hemoglobin



Haematocrit



Mean Corpuscular Volume



Mean Corpuscular Hemoglobin



Mean Corpuscular Hemoglobin Concentration



(WBC)



(RBC)



(HGB)



(HCT)



(MCV)



(MCH)



(MCHC)



(103 cells/µL)



(106 cells/µL)



(g/dL)



(%)



(fL)



(pg)



(g/dL)



G1R-S1, M & 0



Mean



12.73



9.36



16.36



49.50



52.96



17.56



33.12



±SD



2.18



0.62



1.49



1.67



2.94



2.13



3.07



n



5



5



5



5



5



5



5



G2R-S1, M & 0.001



Mean



10.81



9.29



16.20



48.16



51.94



17.46



33.62



±SD



2.36



0.69



1.14



3.27



2.49



0.90



0.57



n



5



5



5



5



5



5



5



G3R-S1, M & 0.003



Mean



8.43



8.10



15.70



41.60



51.46



19.90



38.50



±SD



0.91



1.00



2.82



4.11



1.56



5.41



9.73



n



5



5



5



5



5



5



5



G4R-S1, M & 0.01



Mean



9.05



7.17



14.96



37.62*



52.76



22.42



42.38



±SD



2.17



2.22



1.27



11.03



2.35



6.94



12.54



n



5



5



5



5



5



5



5



G5R-S1, M & 0.03



Mean



9.02



7.47



14.28



39.98



53.54



19.78



37.10



±SD



4.37



1.58



2.69



8.60



1.92



5.16



10.50



n



5



5



5



5



5



5



5



G1R-S1, F & 0



Mean



8.67



8.54



16.06



46.36



54.26



18.86



34.72



±SD



1.14



0.49



0.80



3.04



1.33



1.10



1.51



n



5



5



5



5



5



5



5



G2R-S1, F & 0.001



Mean



10.33



8.55



15.90



45.62



53.38



18.64



34.88



±SD



1.64



0.44



0.70



2.67



1.36



0.36



0.45



n



5



5



5



5



5



5



5



G3R-S1, F & 0.003



Mean



9.24



8.60



16.48



48.10



55.94



19.22



34.34



±SD



1.00



0.45



0.87



2.87



2.27



0.82



0.30



n



5



5



5



5



5



5



5



G4R-S1, F & 0.01



Mean



8.07



8.29



15.92



46.18



55.76



19.24



34.50



±SD



1.26



0.54



0.83



2.97



1.85



0.34



0.77



n



5



5



5



5



5



5



5



G5R-S1, F & 0.03



Mean



10.21



8.71



16.18



47.52



54.58



18.58



34.04



±SD



4.76



0.43



0.66



1.89



0.92



0.23



0.49



n



5



5



5



5



5



5



5



M: Male; F: Female; R: Recovery Group SD: Standard Deviation; n: Number of animals; R: Recovery; S1: Satellite 1; *: Statistically significant (p<0.05).


 


TABLE 10 (Contd…).  SUMMARY OF HAEMATOLOGY RECORD


                                                                                       Refer Appendix 10



























































































































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Platelet Count



Mean Platelet Volume



Reticulocyte Count



Neutrophils



Lymphocytes



Monocytes



Eosinophils



Basophils



Absolute Reticulocyte Count



(PLT)



(MPV)



(Retic)



(Neut)



(Lymph)



(Mono)



(Eos)



(Baso)



(Retic)



(103 cells/µL)



(fL)



(%)



(%)



(%)



(%)



(%)



(%)



(109 cells/L)



G1R-S1, M & 0



Mean



740.40



6.60



2.04



24.54



68.68



3.96



1.66



0.62



189.30



±SD



234.35



1.46



0.50



6.25



6.48



0.35



0.73



0.25



41.55



n



5



5



5



5



5



5



5



5



5



G2R-S1, M & 0.001



Mean



819.20



6.14



2.11



30.92



63.14



3.48



1.72



0.26*



192.50



±SD



211.84



0.38



0.60



5.83



6.26



0.98



0.51



0.09



42.17



n



5



5



5



5



5



5



5



5



5



G3R-S1, M & 0.003



Mean



906.20



6.32



1.69



33.28*



60.40



3.90



1.54



0.22*



136.88



±SD



130.90



0.54



0.17



3.28



3.15



1.01



0.21



0.11



25.25



n



5



5



5



5



5



5



5



5



5



G4R-S1, M & 0.01



Mean



689.60



6.52



2.65



25.36



68.96



3.42



1.28



0.34*



186.32



±SD



106.82



0.50



0.71



4.64



4.68



0.85



0.65



0.11



62.29



n



5



5



5



5



5



5



5



5



5



G5R-S1, M & 0.03



Mean



641.20



6.24



1.78



34.38*



59.38*



4.18



1.30



0.22*



131.18



±SD



99.92



0.26



0.54



5.29



5.66



1.29



0.68



0.08



37.90



n



5



5



5



5



5



5



5



5



5



G1R-S1, F & 0



Mean



1034.20



6.18



1.77



25.76



67.08



4.40



1.62



0.42



150.30



±SD



126.97



0.13



0.58



3.61



5.33



2.06



0.73



0.08



44.91



n



5



5



5



5



5



5



5



5



5



G2R-S1, F & 0.001



Mean



877.20



6.14



1.52



24.88



68.92



3.48



1.62



0.38



129.70



±SD



248.96



0.15



0.32



4.61



4.30



0.75



0.31



0.08



27.19



n



5



5



5



5



5



5



5



5



5



G3R-S1, F & 0.003



Mean



803.40



6.16



1.58



29.02



63.36



4.56



1.90



0.42



135.70



±SD



175.59



0.23



0.35



3.66



3.75



1.48



0.55



0.16



30.58



n



5



5



5



5



5



5



5



5



5



G4R-S1, F & 0.01



Mean



778.00



6.48



1.89



25.88



66.34



3.92



2.72



0.42



156.88



±SD



201.58



0.51



0.44



3.58



2.69



1.08



2.43



0.16



39.65



n



5



5



5



5



5



5



5



5



5



G5R-S1, F & 0.03



Mean



881.80



6.20



1.85



24.62



69.68



2.96



1.66



0.42



161.58



±SD



67.34



0.10



0.23



4.88



5.78



0.98



0.91



0.28



21.32



n



5



5



5



5



5



5



5



5



5



M: Male; F: Female; R: Recovery Group SD: Standard Deviation; n: Number of animals; R: Recovery; S1: Satellite 1; *: Statistically significant (p<0.05).


 


TABLE 10 (Contd…). SUMMARY OF HAEMATOLOGY RECORD


    Refer Appendix 10

























































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Absolute Neutrophils



Absolute Lymphocytes



Absolute Monocytes



Absolute Eosinophils



Absolute Basophils



Prothrombin Time



Activated Prothrombin Time



(Neut)



(Lymph)



(Mono)



(Eos)



(Baso)



(PT)



(APTT)



(103 cells/µL)



(103 cells/µL)



(103 cells/µL)



(103 cells/µL)



(103 cells/µL)



(Seconds)



(Seconds)



G1R-S1, M & 0



Mean



3.16



8.69



0.51



0.22



0.08



21.22



20.40



±SD



1.19



1.37



0.12



0.12



0.04



2.07



2.53



n



5



5



5



5



5



5



5



G2R-S1, M & 0.001



Mean



3.38



6.79



0.37



0.19



0.03*



20.48



21.02



±SD



1.07



1.48



0.13



0.07



0.02



1.40



5.12



n



5



5



5



5



5



5



5



G3R-S1, M & 0.003



Mean



2.80



5.08*



0.34



0.13



0.02*



21.36



22.54



±SD



0.36



0.51



0.12



0.03



0.01



0.65



3.78



n



5



5



5



5



5



5



5



G4R-S1, M & 0.01



Mean



2.29



6.24



0.31



0.13



0.03*



19.64



23.34



±SD



0.64



1.64



0.09



0.08



0.02



1.26



4.86



n



5



5



5



5



5



5



5



G5R-S1, M & 0.03



Mean



3.12



5.29*



0.42



0.11



0.02*



20.14



20.56



±SD



1.71



2.31



0.33



0.06



0.02



1.29



1.45



n



5



5



5



5



5



5



5



G1R-S1, F & 0



Mean



2.26



5.78



0.40



0.14



0.04



22.28



24.16



±SD



0.62



0.46



0.25



0.06



0.01



2.77



8.18



n



5



5



5



5



5



5



5



G2R-S1, F & 0.001



Mean



2.53



7.15



0.36



0.17



0.04



18.88



28.28



±SD



0.42



1.44



0.09



0.06



0.02



2.74



7.23



n



5



5



5



5



5



5



5



G3R-S1, F & 0.003



Mean



2.68



5.84



0.43



0.17



0.04



19.64



27.48



±SD



0.44



0.64



0.18



0.04



0.02



2.52



1.82



n



5



5



5



5



5



5



5



G4R-S1, F & 0.01



Mean



2.08



5.37



0.32



0.21



0.03



17.32*



21.16



±SD



0.36



0.95



0.13



0.16



0.02



1.97



6.80



n



5



5



5



5



5



5



5



G5R-S1, F & 0.03



Mean



2.48



7.13



0.33



0.14



0.05



17.44*



25.28



±SD



1.17



3.31



0.28



0.04



0.07



2.74



2.73



n



5



5



5



5



5



5



5



M: Male; F: Female; R: Recovery Group SD: Standard Deviation; n: Number of animals; R: Recovery; S1: Satellite 1; *: Statistically significant (p<0.05)


 


 


TABLE 10 (Contd…).  SUMMARY OF HAEMATOLOGY RECORD


                                                                                       Refer Appendix 10

























































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Total


Leucocyte Count



Total


Erythrocyte Count



Hemoglobin



Haematocrit



Mean Corpuscular Volume



Mean Corpuscular Hemoglobin



Mean Corpuscular Hemoglobin Concentration



(WBC)



(RBC)



(HGB)



(HCT)



(MCV)



(MCH)



(MCHC)



(103 cells/µL)



(106 cells/µL)



(g/dL)



(%)



(fL)



(pg)



(g/dL)



G1R-S2, M & 0



Mean



10.12



8.94



15.38



45.86



51.36



17.18



33.46



±SD



3.50



0.84



1.58



4.30



1.76



0.75



0.43



n



5



5



5



5



5



5



5



G2R-S2, M & 0.001



Mean



8.21



9.28



16.56



48.54



52.32



17.84



34.08



±SD



1.21



0.25



0.23



1.69



0.90



0.35



0.86



n



5



5



5



5



5



5



5



G3R-S2, M & 0.003



Mean



9.34



8.78



15.98



46.76



53.30



18.20



34.18



±SD



0.95



0.37



0.04



0.88



3.01



0.77



0.66



n



5



5



5



5



5



5



5



G4R-S2, M & 0.01



Mean



9.82



9.18



16.26



47.42



51.66



17.74



34.34



±SD



1.92



0.38



0.69



2.35



1.47



0.59



0.65



n



5



5



5



5



5



5



5



G5R-S2, M & 0.03



Mean



8.31



9.18



16.52



48.86



53.26



18.02



33.80



±SD



1.52



0.49



0.41



1.83



1.77



0.90



0.65



n



5



5



5



5



5



5



5



G1R-S2, F & 0



Mean



6.18



8.28



15.94



45.04



54.42



19.28



35.38



±SD



0.42



0.31



0.43



2.22



1.60



0.34



0.82



n



5



5



5



5



5



5



5



G2R-S2, F & 0.001



Mean



8.84*



8.53



16.22



45.92



53.84



19.02



35.32



±SD



2.09



0.22



0.73



1.79



1.52



0.51



0.63



n



5



5



5



5



5



5



5



G3R-S2, F & 0.003



Mean



6.87



8.15



15.32



44.18



54.24



18.84



34.74



±SD



1.58



0.32



0.36



1.18



1.02



0.34



0.25



n



5



5



5



5



5



5



5



G4R-S2, F & 0.01



Mean



7.02



8.47



15.78



45.78



54.08



18.62



34.42



±SD



0.68



0.40



0.82



2.49



1.16



0.72



0.77



n



5



5



5



5



5



5



5



G5R-S2, F & 0.03



Mean



6.71



8.08



15.58



44.20



54.68



19.28



35.26



±SD



0.62



0.29



1.06



2.92



1.98



0.82



0.80



n



5



5



5



5



5



5



5



M: Male; F: Female; R: Recovery Group SD: Standard Deviation; n: Number of animals; R; Recovery: S2: Satellite 2; *: Statistically significant (p<0.05)


 


TABLE 10 (Contd…).  SUMMARY OF HAEMATOLOGY RECORD


                                                                                       Refer Appendix 10



























































































































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Platelet Count



Mean Platelet Volume



Reticulocyte Count



Neutrophils



Lymphocytes



Monocytes



Eosinophils



Basophils



Absolute Reticulocyte Count



(PLT)



(MPV)



(Retic)



(Neut)



(Lymph)



(Mono)



(Eos)



(Baso)



(Retic)



(103 cells/µL)



(fL)



(%)



(%)



(%)



(%)



(%)



(%)



(109 cells/L)



G1R-S2, M & 0



Mean



1028.00



7.22



2.61



28.52



63.52



3.98



2.88



0.26



230.08



±SD



129.94



0.30



0.84



2.97



1.45



1.47



0.44



0.09



64.54



n



5



5



5



5



5



5



5



5



5



G2R-S2, M & 0.001



Mean



982.20



7.26



1.83



31.78



60.88



4.20



2.06



0.32



170.62



±SD



224.15



0.44



0.46



7.04



6.91



1.05



0.69



0.13



47.20



n



5



5



5



5



5



5



5



5



5



G3R-S2, M & 0.003



Mean



1039.40



7.22



2.30



28.88



61.36



4.58



4.02



0.20



200.44



±SD



258.52



0.13



0.61



8.32



7.90



1.41



1.97



0.10



46.56



n



5



5



5



5



5



5



5



5



5



G4R-S2, M & 0.01



Mean



1198.80



7.30



2.24



35.12



56.86



4.08



2.88



0.24



205.74



±SD



247.11



0.10



0.30



6.73



6.15



0.61



0.78



0.09



30.44



n



5



5



5



5



5



5



5



5



5



G5R-S2, M & 0.03



Mean



985.80



7.22



1.87



31.04



60.86



4.08



2.84



0.24



172.20



±SD



236.52



0.28



0.79



5.91



5.79



1.07



0.99



0.05



76.37



n



5



5



5



5



5



5



5



5



5



G1R-S2, F & 0



Mean



848.00



7.46



2.09



33.12



59.26



4.30



2.24



0.20



173.46



±SD



210.18



0.18



0.46



8.08



8.13



0.82



0.72



0.07



39.98



n



5



5



5



5



5



5



5



5



5



G2R-S2, F & 0.001



Mean



963.00



7.32



1.89



28.90



62.82



4.80



2.30



0.24



161.26



±SD



188.85



0.35



0.17



7.23



7.61



0.63



0.89



0.09



17.93



n



5



5



5



5



5



5



5



5



5



G3R-S2, F & 0.003



Mean



1033.00



7.30



1.95



33.08



58.74



5.34



1.68



0.24



159.02



±SD



214.43



0.12



0.33



5.55



4.94



1.74



0.82



0.05



25.82



n



5



5



5



5



5



5



5



5



5



G4R-S2, F & 0.01



Mean



1014.40



7.40



1.94



31.28



61.94



3.90



1.94



0.16



163.24



±SD



147.21



0.28



0.47



6.29



5.09



1.33



0.29



0.05



35.86



n



5



5



5



5



5



5



5



5



5



G5R-S2, F & 0.03



Mean



951.20



7.56



2.07



31.78



60.54



4.44



1.98



0.24



168.48



±SD



162.41



0.15



0.74



7.59



6.65



1.91



0.39



0.09



64.86



n



5



5



5



5



5



5



5



5



5



M: Male; F: Female; R: Recovery Group SD: Standard Deviation; n: Number of animals; R: Recovery; S2: Satellite 2.


 


TABLE 10 (Contd…). SUMMARY OF HAEMATOLOGY RECORD


    Refer Appendix 10

























































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Absolute Neutrophils



Absolute Lymphocytes



Absolute Monocytes



Absolute Eosinophils



Absolute Basophils



Prothrombin Time



Activated Prothrombin Time



(Neut)



(Lymph)



(Mono)



(Eos)



(Baso)



(PT)



(APTT)



(103 cells/µL)



(103 cells/µL)



(103 cells/µL)



(103 cells/µL)



(103 cells/µL)



(Seconds)



(Seconds)



G1R-S2, M & 0



Mean



2.84



6.46



0.42



0.28



0.03



23.42



24.82



±SD



0.93



2.31



0.21



0.09



0.02



1.28



9.26



n



5



5



5



5



5



5



5



G2R-S2, M & 0.001



Mean



2.58



5.01



0.35



0.17



0.03



21.16



18.84



±SD



0.54



1.08



0.11



0.07



0.01



1.42



3.31



n



5



5



5



5



5



5



5



G3R-S2, M & 0.003



Mean



2.71



5.71



0.43



0.38



0.02



22.64



21.20



±SD



0.82



0.74



0.17



0.20



0.01



2.05



4.39



n



5



5



5



5



5



5



5



G4R-S2, M & 0.01



Mean



3.38



5.64



0.40



0.28



0.02



22.82



22.44



±SD



0.59



1.61



0.12



0.08



0.02



2.08



8.71



n



5



5



5



5



5



5



5



G5R-S2, M & 0.03



Mean



2.52



5.11



0.34



0.25



0.02



20.36*



22.64



±SD



0.31



1.34



0.08



0.12



0.01



1.60



6.46



n



5



5



5



5



5



5



5



G1R-S2, F & 0



Mean



2.04



3.67



0.27



0.14



0.01



22.48



24.98



±SD



0.49



0.55



0.05



0.05



0.00



0.86



6.54



n



5



5



5



5



5



5



5



G2R-S2, F & 0.001



Mean



2.62



5.50*



0.43



0.19



0.02



21.04



18.10*



±SD



1.15



1.13



0.13



0.06



0.01



1.96



2.62



n



5



5



5



5



5



5



5



G3R-S2, F & 0.003



Mean



2.25



4.05



0.38



0.11



0.02



21.26



20.70



±SD



0.49



1.04



0.23



0.04



0.01



1.90



2.19



n



5



5



5



5



5



5



5



G4R-S2, F & 0.01



Mean



2.22



4.33



0.27



0.14



0.01



19.34



19.22



±SD



0.64



0.34



0.08



0.02



0.00



4.27



1.31



n



5



5



5



5



5



5



5



G5R-S2, F & 0.03



Mean



2.11



4.08



0.30



0.13



0.01



17.14*



21.26



±SD



0.42



0.71



0.14



0.02



0.00



4.53



1.89



n



5



5



5



5



5



5



5



M: Male; F: Female; R: Recovery Group SD: Standard Deviation; n: Number of animals; R: Recovery; S2: Satellite 2; *: Statistically significant (p<0.05)


 



  • SUMMARY OF BALF RECORD


                                                                                                                                                                                             Refer Appendix 11

























































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Total Leucocyte Count



Neutrophils



Lymphocytes



Monocytes



Eosinophils



Basophils



Absolute Neutrophils



(WBC)



(Neut)



(Lymph)



(Mono)



(Eos)



(Baso)



(Neut)



(103 cells/µL)



(%)



(%)



(%)



(%)



(%)



(103 cells/µL)



G1, M & 0



Mean



0.88



21.73



59.78



2.67



3.54



3.02



0.24



±SD



0.95



10.33



12.31



1.92



8.94



4.13



0.41



n



10



10



10



10



10



10



10



G2, M & 0.001



Mean



0.36



25.22



64.44



2.22



1.65



4.22



0.09



±SD



0.19



8.23



11.95



1.83



1.48



2.60



0.03



n



10



10



10



10



10



10



10



G3, M & 0.003



Mean



0.51



34.81*



54.66



4.38



3.02



3.47



0.20



±SD



0.36



12.14



17.33



4.70



1.32



1.50



0.22



n



10



10



10



10



10



10



10



G4, M & 0.01



Mean



0.43



27.22



62.94



2.66



2.23



4.49



0.12



±SD



0.11



12.56



18.13



1.50



1.30



3.03



0.07



n



10



10



10



10



10



10



10



G5, M & 0.03



Mean



0.44



23.92



70.22



2.15



2.75



5.14



0.09



±SD



0.24



12.01



13.55



1.16



1.72



2.97



0.05



n



10



10



10



10



10



10



10



G1, F & 0



Mean



0.61



16.93



75.39



2.56



3.43



9.87



0.12



±SD



0.32



9.08



11.06



1.40



2.89



5.02



0.10



n



10



10



10



10



10



10



10



G2, F & 0.001



Mean



0.13*



21.33



62.58



0.35*



15.55*



12.95



0.02*



±SD



0.06



19.45



30.57



0.62



14.61



4.61



0.01



n



10



10



10



10



10



10



10



G3, F & 0.003



Mean



0.21*



8.86



83.92



0.34*



6.81



13.04



0.02*



±SD



0.10



4.16



7.77



0.58



3.96



3.52



0.01



n



10



10



10



10



10



10



10



G4, F & 0.01



Mean



0.21*



9.94



86.14



0.38*



3.50



13.86



0.02*



±SD



0.09



5.16



6.46



0.34



2.62



4.77



0.01



n



10



10



10



10



10



10



10



G5, F & 0.03



Mean



0.18*



14.98



78.18



0.42*



6.28



11.09



0.02*



±SD



0.11



8.26



9.85



0.48



3.30



4.69



0.01



n



10



10



10



10



10



10



10



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; *: Statistically significant (p<0.05)


TABLE 11 (Contd…). SUMMARY OF BALF RECORD


        Refer Appendix 11


























































































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Absolute Lymphocytes



Absolute Monocytes



Absolute Eosinophils



Absolute Basophils



Lactate Dehydrogenase



Total


Protein



Alkaline phosphatase



Gamma-glutamyl transferase



(Lymph)



(Mono)



(Eos)



(Baso)



(LDH)



(TPR)



(ALP)



(GGT)



(103 cells/µL)



(103 cells/µL)



(103 cells/µL)



(103 cells/µL)



(u/L)



(g/dL)/(µg/ml)



(U/L)



(U/L)



G1, M & 0



Mean



0.50



0.04



0.03



0.02



68.50



0.43



10.80



3.45



±SD



0.47



0.07



0.07



0.02



47.49



0.26



2.53



2.46



n



10



10



10



10



10



10



10



10



G2, M & 0.001



Mean



0.24



0.01



0.01



0.02



59.83



0.24*



10.00



3.88



±SD



0.16



0.01



0.01



0.01



27.87



0.05



0.00



2.77



n



10



10



10



10



10



10



10



10



G3, M & 0.003



Mean



0.24



0.04



0.02



0.01



56.66



0.21*



11.30



2.21



±SD



0.08



0.07



0.02



0.01



49.34



0.09



4.11



0.76



n



10



10



10



10



10



10



10



10



G4, M & 0.01



Mean



0.27



0.01



0.01



0.02



38.12



0.30



10.00



1.58



±SD



0.12



0.01



0.01



0.02



9.74



0.10



0.00



1.06



n



10



10



10



10



10



10



10



10



G5, M & 0.03



Mean



0.33



0.01



0.01



0.02



61.19



0.32



10.00



3.19



±SD



0.23



0.01



0.01



0.02



35.37



0.13



0.00



2.16



n



10



10



10



10



10



10



10



10



G1, F & 0



Mean



0.45



0.02



0.02



0.07



207.67



294.00



20.80



3.62



±SD



0.24



0.01



0.01



0.05



200.35



136.52



20.51



1.64



n



10



10



10



10



10



10



10



10



G2, F & 0.001



Mean



0.09*



0.00*



0.02



0.02*



42.75*



272.37



10.00



2.58



±SD



0.07



0.00



0.01



0.01



30.45



61.71



0.00



0.72



n



10



10



10



10



10



10



10



10



G3, F & 0.003



Mean



0.18*



0.00*



0.01



0.03*



87.84



233.41



10.10



2.58



±SD



0.10



0.00



0.01



0.02



63.93



78.80



0.32



2.66



n



10



10



10



10



10



10



10



10



G4, F & 0.01



Mean



0.18*



0.00*



0.01



0.03*



140.33



290.17



14.90



9.73



±SD



0.08



0.00



0.01



0.02



84.92



151.87



10.46



20.03



n



10



10



10



10



10



10



10



10



G5, F & 0.03



Mean



0.14*



0.00*



0.01



0.02*



91.54



304.69



10.50



3.74



±SD



0.09



0.00



0.01



0.02



61.16



82.45



0.71



2.19



n



10



10



10



10



10



10



10



10



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; *: Statistically significant (p<0.05)


TABLE 11 (Contd…). SUMMARY OF BALF RECORD


        Refer Appendix 11

























































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Total Leucocyte Count



Neutrophils



Lymphocytes



Monocytes



Eosinophils



Basophils



Absolute Neutrophils



(WBC)



(Neut)



(Lymph)



(Mono)



(Eos)



(Baso)



(Neut)



(103 cells/µL)



(%)



(%)



(%)



(%)



(%)



(103 cells/µL)



G1R-S1, M & 0



Mean



1.15



34.48



50.78



3.66



3.32



1.72



0.51



±SD



1.34



17.34



18.00



2.99



3.45



1.14



0.78



n



5



5



5



5



5



5



5



G2R-S1, M & 0.001



Mean



0.91



24.16



68.50



2.72



0.92



3.02



0.27



±SD



0.70



19.70



19.58



3.40



0.54



1.22



0.34



n



5



5



5



5



5



5



5



G3R-S1, M & 0.003



Mean



0.73



26.20



67.46



2.04



1.18



4.42



0.19



±SD



0.18



15.92



17.29



1.07



0.71



4.11



0.11



n



5



5



5



5



5



5



5



G4R-S1, M & 0.01



Mean



1.09



21.50



62.64



11.56



2.78



7.80*



0.36



±SD



1.51



10.82



31.74



22.74



2.97



5.11



0.65



n



5



5



5



5



5



5



5



G5R-S1, M & 0.03



Mean



0.71



26.78



66.42



2.24



2.40



6.18



0.21



±SD



0.21



11.71



12.01



1.50



0.99



3.61



0.16



n



5



5



5



5



5



5



5



G1R-S1, F & 0



Mean



0.46



15.36



73.40



6.44



2.92



11.46



0.13



±SD



0.40



19.36



29.76



12.56



2.24



5.53



0.25



n



5



5



5



5



5



5



5



G2R-S1, F & 0.001



Mean



0.55



17.24



76.60



1.96



2.38



4.38*



0.10



±SD



0.26



16.00



18.12



1.52



2.23



2.81



0.12



n



5



5



5



5



5



5



5



G3R-S1, F & 0.003



Mean



0.35



12.76



81.82



1.14



2.68



8.64



0.04



±SD



0.16



3.92



6.56



0.63



1.73



1.02



0.02



n



5



5



5



5



5



5



5



G4R-S1, F & 0.01



Mean



0.25



14.66



77.64



1.64



3.80



5.42*



0.03



±SD



0.10



4.71



9.14



0.78



3.17



2.78



0.01



n



5



5



5



5



5



5



5



G5R-S1, F & 0.03



Mean



0.56



9.34



86.24



0.68



2.48



7.48



0.05



±SD



0.26



3.08



4.90



0.43



1.20



3.28



0.04



n



5



5



5



5



5



5



5



M: Male; F: Female; SD: Standard Deviation; n: Number of animals; R: Recovery; S1: Satellite 1; *: Statistically significant (p<0.05)


TABLE 11 (Contd…). SUMMARY OF BALF RECORD


        Refer Appendix 11


























































































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Absolute Lymphocytes



Absolute Monocytes



Absolute Eosinophils



Absolute Basophils



Lactate Dehydrogenase



Total


Protein



Alkaline phosphatase



Gamma-glutamyl transferase



(Lymph)



(Mono)



(Eos)



(Baso)



(LDH)



(TPR)



(ALP)



(GGT)



(103 cells/µL)



(103 cells/µL)



(103 cells/µL)



(103 cells/µL)



(u/L)



(g/dL)



(U/L)



(U/L)



G1R-S1, M & 0



Mean



0.50



0.07



0.02



0.02



146.40



0.92



12.20



3.71



±SD



0.43



0.12



0.01



0.02



113.31



1.19



4.92



1.44



n



5



5



5



5



5



5



5



5



G2R-S1, M & 0.001



Mean



0.56



0.04



0.01



0.03



106.30



0.36



16.60



2.69



±SD



0.35



0.08



0.00



0.02



67.47



0.08



10.48



0.81



n



5



5



5



5



5



5



5



5



G3R-S1, M & 0.003



Mean



0.49



0.01



0.01



0.04



73.86



0.26



10.80



3.15



±SD



0.17



0.01



0.01



0.05



21.60



0.10



1.79



1.48



n



5



5



5



5



5



5



5



5



G4R-S1, M & 0.01



Mean



0.31



0.40



0.01



0.04



131.90



0.53



13.40



4.02



±SD



0.13



0.88



0.01



0.01



122.12



0.39



5.98



1.44



n



5



5



5



5



5



5



5



5



G5R-S1, M & 0.03



Mean



0.45



0.02



0.01



0.04



96.10



0.32



17.60



4.64



±SD



0.08



0.02



0.01



0.03



50.22



0.14



11.28



2.74



n



5



5



5



5



5



5



5



5



G1R-S1, F & 0



Mean



0.24



0.07



0.01



0.05



67.24



0.29



19.00



1.65



±SD



0.06



0.15



0.00



0.04



68.82



0.15



19.03



1.11



n



5



5



5



5



5



5



5



5



G2R-S1, F & 0.001



Mean



0.43



0.01



0.01



0.02



57.12



0.34



13.40



1.32



±SD



0.25



0.01



0.00



0.02



35.09



0.18



5.46



0.31



n



5



5



5



5



5



5



5



5



G3R-S1, F & 0.003



Mean



0.30



0.00



0.01



0.03



44.38



0.28



10.60



2.69



±SD



0.15



0.01



0.00



0.01



16.75



0.19



0.89



1.37



n



5



5



5



5



5



5



5



5



G4R-S1, F & 0.01



Mean



0.20



0.00



0.01



0.02



37.94



0.22



10.40



1.75



±SD



0.10



0.00



0.01



0.01



16.45



0.09



0.89



0.62



n



5



5



5



5



5



5



5



5



G5R-S1, F & 0.03



Mean



0.48



0.00



0.01



0.04



60.96



0.20



10.40



2.14



±SD



0.22



0.00



0.01



0.03



22.41



0.05



0.89



0.96



n



5



5



5



5



5



5



5



5



M: Male; F: Female; R: Recovery Group; SD: Standard Deviation; n: Number of animals; R: Recovery; S1: Satellite 1.


 


TABLE 11 (Contd…). SUMMARY OF BALF RECORD


        Refer Appendix 11

























































































































































































































































































































































Group, Sex & Dose (mg/L)


 

Total Leucocyte Count



Neutrophils



Lymphocytes



Monocytes



Eosinophils



Basophils



Absolute Neutrophils



(WBC)



(Neut)



(Lymph)



(Mono)



(Eos)



(Baso)



(Neut)



(103 cells/µL)



(%)



(%)



(%)



(%)



(%)



(103 cells/µL)



G1R-S2, M & 0



Mean



2.03



32.16



58.36



7.16



0.74



3.58



0.81



±SD



2.26



11.48



14.75



5.49



0.23



1.40



1.12



n



5



5



5



5



5



5



5



G2R-S2, M & 0.001



Mean



0.92



36.14



54.90



3.56



1.24



5.38



0.44



±SD



0.76



24.87



27.60



3.77



0.65



3.81



0.60



n



5



5



5



5



5



5



5



G3R-S2, M & 0.003



Mean



0.81



29.26



64.74



2.46



1.56



5.36



0.30



±SD



0.53



16.86



17.46



1.21



1.17



3.09



0.39



n



5



5



5



5



5



5



5



G4R-S2, M & 0.01



Mean



0.48



22.96



72.52



2.02



1.22



5.72



0.17



±SD



0.43



17.64



18.79



0.98



0.55



2.25



0.28



n



5



5



5



5



5



5



5



G5R-S2, M & 0.03



Mean



0.77



29.08



61.40



4.76



2.18



8.72



0.27



±SD



0.48



13.04



12.92



2.26



1.28



6.51



0.29



n



5



5



5



5



5



5



5



G1R-S2, F & 0



Mean



0.31



21.18



69.36



3.06



3.12



6.44



0.07



±SD



0.11



7.76



8.34



0.94



1.25



1.52



0.05



n



5



5



5



5



5



5



5



G2R-S2, F & 0.001



Mean



0.60



21.12



64.72



7.86



4.26



6.46



0.14



±SD



0.47



10.36



22.74



12.22



4.85



3.84



0.14



n



5



5



5



5



5



5



5



G3R-S2, F & 0.003



Mean



0.53



17.46



75.60



4.12



1.24



9.68



0.11



±SD



0.34



9.06



10.34



2.42



0.74



3.62



0.11



n



5



5



5



5



5



5



5



G4R-S2, F & 0.01



Mean



0.35



19.20



72.42



3.32



3.82



9.16



0.07



±SD



0.09



10.86



13.92



2.21



2.01



2.63



0.04



n



5



5



5



5



5



5



5



G5R-S2, F & 0.03



Mean



0.41



18.18



74.92



2.56



2.98



9.78



0.07



±SD



0.14



6.16



7.14



0.47



2.05



1.65



0.03



n



5



5



5



5



5



5



5



M: Male; F: Female; R: Recovery Group; SD: Standard Deviation; n: Number of animals; R: Recovery; S2: Satellite 2.


TABLE 11 (Contd…). SUMMARY OF BALF RECORD


        Refer Appendix 11



































Group, Sex & Dose (mg/L)


 

Absolute Lymphocytes



Absolute Monocytes



Absolute Eosinophils



Absolute Basophils



Lactate Dehydrogenase



Total


Protein



Alkaline phosphatase



Gamma-glutamyl transferase



(Lymph)



(Mono)



(Eos)



(Baso)



(LDH)



(TPR)



(ALP)



(GGT)



(103 cells/µL)



(103 cells/µL)



(103 cells/µL)



(103 cells/µL)



(u/L)



(µg/ml)



(U/L)