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EC number: 947-744-0 | CAS number: -
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
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- Flash point
- Auto flammability
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- Explosiveness
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- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
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- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
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- Endpoint summary
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- Environmental data
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- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
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- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
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- Genetic toxicity
- Carcinogenicity
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- Specific investigations
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- Additional toxicological data
Endpoint summary
Administrative data
Key value for chemical safety assessment
Effects on fertility
Description of key information
In order to assess the fertility properties of the test substance Hostacerin DGSB after repeated oral treatment for a subchronic period, one dose-range-finding and one main OECD 421 screening study is available.
A total of 96 (48 males + 48 females) Sprague Dawley rats were distributed to four groups. Each group (G1, G2, G3and G4) consisted of 12 males and 12 females. The animals in G1 group were administered with vehicle[0.5% w/v Carboxy Methyl Cellulose],animals in G2, G3 and G4groups were administered withtest itemat dose levels of100, 300 and 1000 mg/kg body weight for low dose, mid dose and high dose groups respectively. The vehicle and test item formulations were administered orally by gavageat the dose volume of 10 mL/kg body weight. Males were treated for two weeks pre-mating, during mating and up to the day before sacrifice during post-mating period (total of 34 days of treatment). The females were treated for two weeks pre-mating period, during mating, pregnancy (gestation) and up to lactation day 13 after which the pups were sacrificed on lactation Day 13 and females (dams) were sacrificed on lactation day 14 after overnight fasting (water allowed).
All animals did not reveal any clinical signs of toxicity, no mortality or morbidity observed throughout the experimental period. No treatment related changes in body weight, percent change in body weight, feed consumption, organ weights (both absolute and relative) were observed in both the sex of all tested dose group animals. There were no treatment related changes observed in serum T4 levels of adult animals and in serum T4 levels of pups at any of the tested dose groups.
There were no treatment-related effects on the mean pre-coital time, mating and fertility indices of sires and dams at all the doses tested.Females did not reveal any treatment related changes in oestrus cyclicity, copulatory interval, body weights and feed consumption during gestation and lactation periods, mean gestation length, implantation index, pre-implantation loss, post-implantation loss and number of pups delivered when compared with vehicle control group. No effects were observed for live birth index of each litter, sex ratio and pup survival index at all the tested dose groups throughout the lactation periodwhen compared with vehicle control group animals.
All pups did not reveal any clinical signs or external anomalies throughout the lactation period. No treatment related changes in pup weights, anogenital distance ratio were noted. No occurrences of nipples in male pups at any of the tested dose groups and vehicle control group.
All animals of all dose groups did not reveal any treatment related gross pathological and histopathological findings compared to vehicle control group.
Link to relevant study records
- Endpoint:
- screening for reproductive / developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- 19 May 2017 to 13 October 2017
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Study is a dose range finding study.
- Remarks:
- 6 animals /sex/dose were used
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
- Version / remarks:
- adopted on 29 July 2016
- Deviations:
- yes
- Remarks:
- Study is a dose-range finding study. 6 rats /sex and dose were used.
- Principles of method if other than guideline:
- Study is performed as dose-range-finding study. Thus, only 6 rats per sex and dose are used.
- GLP compliance:
- no
- Limit test:
- no
- Justification for study design:
- SPECIFICATION OF STUDY DESIGN:
- Premating exposure duration for parental (P0) animals: 14 days
- Basis for dose level selection: The doses of 100, 300 and 1000 mg/kg body weight were selected as low (G2), mid (G3) and high dose (G4) levels based on the NOAEL of test item Hostacerin DGSB was found to be 1000 mg/kg/day when administered for a period of 14 consecutive days by oral (gavage) to Sprague Dawley rats obtained from the dose range finding study (Bioneeds Study Number: BIO-TX 2503). I
- Route of administration: Oral gavage
- Other considerations:
choice of species, strain: Rat & Sprague Dawley
vehicle and number of animals: 0.5 % Carboxy methyl cellulose and 6 per group per sex - Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source of test material : Clariant Produkte(Deutschland) GmbH84504 Burgkirchen, Germany.
- Batch No.of test material: DEG4338635
- Expiration date of thebatch:04 Oct 2018
- Purity test date:04 Oct 2016
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Ambient (21 to 29 oC)
- Stability under test conditions: not established (freshly prepared test item formulations were administered to the animals and homogeneity was achieved thorough stirring).
- Solubility in the solvent/vehicle: forms uniform suspension in 0.5% w/v carboxy methyl cellulose at the concentration of 100 mg/mL (highest dose concentration) as evidenced by thein-house solubility/suspendibilty test results.
TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: ground and made in to suspension
- Final preparation of a solid: Suspension
FORM AS APPLIED IN THE TEST (if different from that of starting material): Suspension - Species:
- rat
- Strain:
- Sprague-Dawley
- Details on species / strain selection:
- Rat is one of the standard laboratory rodent species used for toxicity assessment and also recommended by various regulatory authorities.
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source:In-house bred
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 10 weeks
- Weight at study initiation: Males: 225.13 g to 254.32 g; Females: 201.03 g to 221.19 g
- Housing: Animals were housed in a standard polypropylene cage (size: L 430 x B 285 x H 150 mm) with stainless steel mesh top grill having facilities for holding pelleted food and drinking water in water bottle fitted with stainless steel sipper tube. Clean sterilized paddy husk was provided as bedding material.
i. Pre mating
Two animals of same sex and group per cage were housed.
ii. Mating
During mating, two animals (one male and one female) of same group were housed.
iii. Post mating
After confirming presence of sperm in the vaginal smear (Day 0 of pregnancy), the mated pairs were separated. Males were housed with their former cage mates while females were housed individually. Sterilized paper shreds were provided as a nesting material from gestation day 20 onwards.
- Diet (ad libitum): Altromin Maintenance diet for rats and mice 1324 manufactured by Altromin Spezialfutter GmbH & Co. KG was provided ad libitum.
- Water (ad libitum):Water was provided ad libitum throughout the experimental period. Deep bore-well water passed through activated charcoal filter and exposed to ultraviolet rays in Aquaguard water filter with purifier was provided in plastic water bottles with stainless steel sipper tubes.
- Acclimation period:19 May 2017 to 06 June 2017
ENVIRONMENTAL CONDITIONS
- Temperature (°C):19.1 to 24.5
- Humidity (%):41 to 69
- 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: 19 May 2017 To:05 August 2017 - Route of administration:
- oral: gavage
- Remarks on MMAD:
- not applicable
- Vehicle:
- CMC (carboxymethyl cellulose)
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS:
The required quantity of test item grind well in a mortar and in a mortar and pestle by adding small quantity of 0.5% w/v Carboxy Methyl Cellulose untilaclear suspension was formed. Thereafter, the entire quantity of the formulation was transferred intoa measuring cylinder. Again,a small quantity of the vehicle was added to rinse the mortar and pestleand this was transferred into the measuring cylinder. The rinsing of mortar and pestle was repeated to ensure the complete transfer of the contents to the measuring cylinder. Finally, the volume was made up to the required quantity with vehicle to get a desired concentration of 10, 30 and 100 mg/mL of test item for low, mid and high dose groups respectively. - Details on mating procedure:
- - M/F ratio per cage: 1:1
- Length of cohabitation: 14 days
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy
- Further matings after two unsuccessful attempts: no
- After successful mating each pregnant female was caged : individually
- Any other deviations from standard protocol: no - Analytical verification of doses or concentrations:
- no
- Details on analytical verification of doses or concentrations:
- Stability and homogeneity of the test substance in dose formulation were not established under this dose range findiong study. However, test item formulations were prepared daily fresh.
- Duration of treatment / exposure:
- Males: 30 days (premating, mating and post mating periods).
Females: two weeks pre-mating period, during mating, pregnancy (gestation) and up to lactation day 13. - Frequency of treatment:
- Once daily
- Details on study schedule:
- Age at mating of the mated animals in the study: 14 to 15 weeks
- Dose / conc.:
- 0 mg/kg bw/day (nominal)
- Remarks:
- vehicle control
- Dose / conc.:
- 100 mg/kg bw/day (nominal)
- Remarks:
- low dose
- Dose / conc.:
- 300 mg/kg bw/day (nominal)
- Remarks:
- mid dose
- Dose / conc.:
- 1 000 mg/kg bw/day (nominal)
- Remarks:
- high dose
- No. of animals per sex per dose:
- 6 animals per sex per dose
- Control animals:
- yes
- Details on study design:
- - Dose selection rationale: The doses of 100, 300 and 1000 mg/kg body weight were selected as low (G2), mid (G3) and high dose (G4) levels for test item Hostacerin DGSB based on the No Observed Adverse Effect Level (NOAEL) of the test item Hostacerin DGSB was found to be 1000 mg/kg/day when administered for a period of 14 consecutive days by oral (gavage) to Sprague Dawley rats obtained from the dose range finding study (Bioneeds Study Number: BIO-TX 2503).
- Rationale for animal assignment :The body weight stratified animals were distributed to all the groups using Microsoft Excel Spreadsheet, such that body weight variation of animals selected for the study did not exceed ± 20% (+5.07 and -7.14 for males and +5.59 and -4.96 for females) of the mean body weight of each sex. - Positive control:
- not applicable
- Parental animals: Observations and examinations:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: once daily
- Cage side observations checked in table [No.1] were included.
DETAILED CLINICAL OBSERVATIONS: no
BODY WEIGHT: Yes
- Time schedule for examinations: first day of dosing, weekly thereafter and at termination. The females were weighed on gestation days 0, 7, 14 and 20 during pregnancy and on days 1, 4, 7 and 13 during lactation period.
FOOD CONSUMPTION : Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes; weekly once; gestation days 0 to 7, 7 to 14 and 14 to 20 duringpregnancy and day 1 to 4, 4 to 7, 7 to 13 during lactation period.
- Oestrous cyclicity (parental animals):
- two weeks after five days of acclimatization to evaluate its normal oestrus cyclicity (4 to 5 days), daily from the beginning of the treatment period until evidence of mating and on the day of sacrifice for females.
- Sperm parameters (parental animals):
- Parameters examined Epididymis, Prostate+ Seminal vesicles weight and Testes weight
- Litter observations:
- STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no
PARAMETERS EXAMINED
The following parameters were examined in offspring:
[number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities.
GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death for pups born or found dead. - Postmortem examinations (parental animals):
- SACRIFICE
- Male animals: All animals [after completion of 30 days of treatment]
- Maternal animals: All animals [on lactation day 14]
GROSS NECROPSY
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]
ORGAN WEIGHTS
In Table [5 and 6] weighed and preserved. - Postmortem examinations (offspring):
- SACRIFICE
- The offspring were sacrificed at [table 20] 13 days of age (lactation day 13)
- These animals were subjected to postmortem examinations (macroscopic examination) as follows:
GROSS NECROPSY
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]
- Statistics:
- The data was subjected to various statistical analysis using SPSS software version 22.
All analysis and comparisons were evaluated at the 95% level of confidence (P<0.05)
The statistical analysis was followed to the parameters as mentioned below:
Body weight, Change in body weight, Feed consumption, Copulatory interval, Gestation length, Organ
weights, Mean pup weight, Pre implantation loss, Pre natal loss, Post natal
loss, Post implantation loss:- Parametric - One-way ANOVA with Dunnett’s post test.
Corpora lutea/dam, Implantations/dam, No. of pups/dam, Sex ratio, Litter size, Pup weight:- Non
Parametric - Kruskal-Wallis followed by the Mann-Whitney test.
Pregnancy rate. Dams with live pups, Dams with dead pups, No. of litters with/without dead pups, No.
of litters with/without resorptions:- Cross Tabs - Chi-square test/ Fischer's Exact Test - Reproductive indices:
- Reproductive indices
Male Fertility Index (%) = Number of Males Impregnating Females
------------------------------------------------- x 100
Number of Males used for Mating
Female Fertility Index (%) = Number of Females confirmed with Pregnancy
--------------------------------------------------------- x 100
Number of Females used for Mating
Parturition (%) = Number Of Parturitions
------------------------------- x 100
Number Of Pregnancies
Copulatory Interval (Days) = Date of Initiation of Cohabitation - Date of Confirmation of Mating
Gestation Length (days) = Date of Delivery - Date of Confirmation of Mating - Offspring viability indices:
- Offspring viability indices
Male/Female Sex Ratio = Number Male Pups Per Dam
-----------------------------------
Number Female Pups Per Dam
Live Birth Index (%) = Number Of Viable Pups Born
------------------------------------ x 100
Number Of Pups Born
Lactation Day 4
Pup Survival Index (%) = Number Of Viable Pups on Lactation Day 4
----------------------------------------------------- x 100
Number Of Pups Born
Lactation Day 7
Pup Survival Index (%) = Number Of Viable Pups on Lactation Day 7
------------------------------------------------------ x 100
Number Of Viable Pups on Lactation Day 4
Lactation Day 13 Pup Survival Index (%) = Number Of Viable Pups on Lactation Day 13
------------------------------------------------------- x 100
Number Of Viable Pups on Lactation Day 7 - Clinical signs:
- no effects observed
- Description (incidence and severity):
- There were no clinical signs of toxicity observed at any of the tested dose group animals and vehicle control group animals of either sex during premating period and mating period; during post mating period in males and during gestation/lactation period in females.
- Dermal irritation (if dermal study):
- not examined
- Description (incidence and severity):
- not applicable
- Mortality:
- no mortality observed
- Description (incidence):
- There were no mortality/morbidity observed at any of the tested dose group animals and vehicle control group animals of either sex during premating period and mating period; during post mating period in males and during gestation/lactation period in females.
- Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- There were no treatment related changes in mean body weight and percent change in body weight with respect to day 1 at any of the tested dose group animals of either sex when compared with vehicle control group animals.
- Food consumption and compound intake (if feeding study):
- effects observed, non-treatment-related
- Description (incidence and severity):
- There were no treatment related changes noted in feed consumption at any of the tested dose group animals of either sex when compared with vehicle control group animals. However, statistical significant increase during week 2 in G2 and G3 males; increase during post mating in G2 males; increase during week 1 and 2 in G4 females was noted when compared with vehicle control group animals. This can be considered as incidental due to lack of dose dependency and no changes were noted in body weight during these periods.
There were no changes observed in the feed consumption during gestation period at any of the tested dose group animals when compared with vehicle control group animals. However, statistical significant increase in mean feed consumption during gestation day 0 to 7 was noted at low dose group animals when compared with vehicle control group animals. This change is considered as incidental and not treatment related as there were no effects noted in mean body weight at this duration and the change is inconsistent.
There were no changes observed in the feed consumption during lactation period at any of the tested dose group animals when compared with vehicle control group animals - Food efficiency:
- not examined
- Description (incidence and severity):
- not applicable
- Water consumption and compound intake (if drinking water study):
- not examined
- Description (incidence and severity):
- not applicable
- Ophthalmological findings:
- not examined
- Description (incidence and severity):
- not applicable
- Haematological findings:
- no effects observed
- Description (incidence and severity):
- No treatment related changes were observed in serum T4 levels of adult animals.
- Clinical biochemistry findings:
- not examined
- Description (incidence and severity):
- not applicable
- Urinalysis findings:
- not examined
- Description (incidence and severity):
- not applicable
- Behaviour (functional findings):
- not examined
- Description (incidence and severity):
- not applicable
- Immunological findings:
- not examined
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Histopathological findings: non-neoplastic:
- not examined
- Histopathological findings: neoplastic:
- not examined
- Other effects:
- not examined
- Reproductive function: oestrous cycle:
- no effects observed
- Description (incidence and severity):
- There were no changes observed in the oestrus cyclicity of females at any of the tested doses during pre-mating treatment, mating treatment and on lactation day 14.
- Reproductive function: sperm measures:
- no effects observed
- Description (incidence and severity):
- Testes, which were screened with special emphasis on stages of spermatogenesis and interstitial testicular cell structure, revealed normal progression of the spermatogenic cycle and presence of all expected associations.
- Reproductive performance:
- no effects observed
- Description (incidence and severity):
- No effects were noted in mating index and fertility index of both the males and females at all the tested dose groups
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 1 000 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: No adverse effects observed
- Key result
- Critical effects observed:
- no
- Dermal irritation (if dermal study):
- not examined
- Mortality:
- mortality observed, non-treatment-related
- Body weight and weight changes:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Statistical significant increase in mean pup weight at high dose group (G4) male mean pup weight on lactation day 4 and 7 was noted when compared with vehicle control group. This change is considered as incidental but not treatment related as there were no effects noted in other parameters. No effects were noted in female mean pup weight at this group. The obtained weights are within historical control range.
- Food consumption and compound intake (if feeding study):
- not examined
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- not examined
- Haematological findings:
- no effects observed
- Description (incidence and severity):
- There were no treatment related changes observed in serum T4 levels on lactation day 4 and 13 pups at any of the tested dose groups when compared with vehicle control group
- Clinical biochemistry findings:
- not examined
- Urinalysis findings:
- not examined
- Behaviour (functional findings):
- not examined
- Immunological findings:
- not examined
- Clinical signs:
- no effects observed
- Dermal irritation (if dermal study):
- not examined
- Mortality / viability:
- no mortality observed
- Description (incidence and severity):
- No post natal death observed (at birth until lactation day 13)
- Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- There were no changes observed in mean pup (male and female) weight on lactation day 1, 4, 7 and 13 at any of the tested dose group litters when compared with vehicle control group litters.
- Food consumption and compound intake (if feeding study):
- not examined
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- not examined
- Haematological findings:
- not examined
- Clinical biochemistry findings:
- not examined
- Urinalysis findings:
- not examined
- Sexual maturation:
- not examined
- Organ weight findings including organ / body weight ratios:
- not examined
- Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- There were no gross pathological changes (both external and internal) observed at any of the tested dose group animals of either sex of pups.
- Histopathological findings:
- not examined
- Other effects:
- no effects observed
- Behaviour (functional findings):
- not examined
- Description (incidence and severity):
- No effects were noted in offsprings at all the tested dose groups.
- Key result
- Dose descriptor:
- NOAEL
- Generation:
- F1
- Effect level:
- >= 1 000 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: no adverse effects observed
- Key result
- Critical effects observed:
- no
- Key result
- Reproductive effects observed:
- no
- Lowest effective dose / conc.:
- 1 000 mg/kg bw/day (nominal)
- Treatment related:
- no
- Conclusions:
- Based on the results of the dose-range-finding study discussed, the No Observed Adverse Effect Level (NOAEL) of the test item Hostacerin DGSB was found to be 1000 mg/kg body weight when administered to the males for two weeks pre-mating, during mating and up to the day before sacrifice during post-mating period (total of 30 days) and to the females for two weeks pre-mating, during mating, pregnancy (gestation) and up to lactation day 13 under the experimental conditions employed in this study. Based on the results form the dose range finding study, dose levels of 0, 100, 300 and 1000 mg/kg bw/day are recommended for the main OECD 421 study.
- Executive summary:
The test item, Hostacerin DGSB was evaluated to generate information on health hazards likely to arise from repeated exposure of test item Hostacerin DGSB when administered to male and female Sprague Dawley rats through oral route over a relatively limited period of time and to select the doses for subsequent Reproduction/Developmental Toxicity Screening Test.
A total of 48 (24 males + 24 females) Sprague Dawley rats were distributed to four groups. Each group (G1, G2, G3and G4) consisted of 6 males and 6 females. The animals in G1 group were administered with vehicle [0.5% w/v Carboxy Methyl Cellulose], the animals in G2, G3 and G4groups were administered with test item at the dose levels of100, 300 and 1000 mg/kg body weight for low dose, mid dose and high dose groups respectively. The vehicleand test item formulations were administered orally by gavage at the dose volume of 10 mL/kg body weight.
The males were treated for two weeks pre-mating, during variable period of mating and up to the day before sacrifice during post-mating period (total of 30 days of treatment). The females were treated for two weeks pre-mating period, during variable period of mating, pregnancy (gestation) and up to lactation day 13 after which the pups were sacrificed on lactation Day 13 and females (dams) were sacrificed on lactation day 14 after overnight fasting (water allowed).
All animals were observed for clinical signs, mortality and morbidity, body weight and feed consumption.
Each litter was examined after delivery (lactation day 1) and the number and sex of pups (litter size), dead pups born on day 1 and live births were recorded. Body weights of male and female pups were recorded separately on lactation days 1, 4, 7 and 13.
Gross pathology and organ weighing were performed on day 31 for males and on lactation day 14 for dams. The gross pathology was performed on lactation day 13 for pups. The number of corpora lutea and implantation sites for dams were recorded during necropsy.
The dams were observed for body weight and feed consumption during gestation and lactation period.
The animals did not reveal any clinical signs of toxicity. No mortality or morbidity was observed throughout the experimental period. No treatment related changes in body weight, percent change in body weight, feed consumption and in organ weights (both absolute and relative) were observed in both the sex of all the group animals.
The dams did not reveal any treatment related changes in gestation and lactation body weight, percent change in gestation and lactation body weights, feed consumption during gestation period and lactation period, uteri observations and pup weights.
The adults and lactation day 13 pups did not reveal any treatment related gross pathological findings.
Based on the results form the dose range finding study, dose levels of 0, 100, 300 and 1000 mg/kg bw/day are recommended for the main OECD 421 study.
- Endpoint:
- screening for reproductive / developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2017-09-16 to 2018-04-03
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
- Version / remarks:
- adopted on 29 July 2016
- Deviations:
- no
- Principles of method if other than guideline:
- not applicable
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
- Justification for study design:
- SPECIFICATION OF STUDY DESIGN:
- Premating exposure duration for parental (P0) animals: Yes (14 days)
- Basis for dose level selection: The doses of 0, 100, 300 and 1000 mg/kg body weight for vehicle control, low dose, mid dose and high dose respectively were selected in consultation with the sponsor based on the results of Dose Range Finding Study for Reproduction/Developmental Toxicity Screening Test of Hostacerin DGSB by Oral Gavage in Sprague Dawley Rats (Bioneeds Study No.: BIO-TX 2516).
These doses were selected as the test item Hostacerin DGSB when administered orally to Sprague Dawley males for two weeks prior to mating, during mating and up to the day before sacrifice during post-mating period (total of at 30 days treatment) and females were treated for two weeks pre-mating period, during mating, pregnancy (gestation) and up to lactation day 13 after which the pups were sacrificed on lactation day 13, females (dams) were sacrificed on lactation day 14.
- Route of administration: oral (gavage) route
- Other considerations:
choice of species and strain: Rat (Rattus norvegicus) and Sprague Dawley (standard laboratory rodent species and strain used for toxicity assessment and also recommended by various regulatory authorities)
vehicle: The test item was in-soluble in distilled water and forms clear homogenous suspension in 0.5 % w/v Carboxy Methyl Cellulose at the concentration of 100 mg/mL (highest dose concentration) as evidenced by the in-house solubility and suspendibility test results. Hence, 0.5 % w/v Carboxy Methyl Cellulose was selected for the test item formulation preparation.
number of animals: 96 (48 Males + 48 Females); 12 Males + 12 Females/group as per OECD 421 guideline specification - Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source and batch No.of test material: Clariant Produkte (Deutschland) GmbH
84504 Burgkirchen, Germany and DEG4338635
- Expiration date of the batch: 04.10.2018
- Purity test date: 04.10.2016
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Ambient (21 to 29°C)
- Stability under test conditions: The stability of the test item in dose formulations was established under Bioneeds Study No.: BIO-ANM 450. The test item formulations were stable for 6 hours at room temperature and for 24 hours at 2 to 8 oC at the concentrations of 10 mg/mL and 100 mg/mL.
- Solubility of the test substance in the solvent/vehicle: 0.5 % w/v Carboxy Methyl Cellulose was used as vehicle for the preparation of formulations as the test item formed clear homogenous suspension in 0.5 % w/v Carboxy Methyl Cellulose at the concentration of 100 mg/mL (highest dose concentration) as evidenced by the in-house solubility and suspendibility test results.
FORM AS APPLIED IN THE TEST: Liquid (Suspension) - Species:
- rat
- Strain:
- Sprague-Dawley
- Details on species / strain selection:
- Rat is one of the standard laboratory rodent species used for toxicity assessment and also recommended by various regulatory authorities.
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: In-house bred animals
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 10 weeks
- Weight at study initiation: Males: 220.17 g to 255.97 g; Females: 201.51 g to 231.96 g
- Housing: Animals were housed in a standard polypropylene cage (size: L 430 x B 285 x H 150 mm) with stainless steel mesh top grill having facilities for holding pelleted food and drinking water in water bottle fitted with stainless steel sipper tube. Clean sterilized paddy husk was provided as bedding material.
i. Pre mating
Two animals of same sex and group per cage were housed.
ii. Mating
During mating, two animals (one male and one female) of same group were housed.
iii. Post mating
After confirming presence of sperm in the vaginal smear (Day 0 of pregnancy), the mated pairs were separated. Males were housed with their former cage mates while females were housed individually. Sterilized paper shreds were provided as a nesting material from gestation day 20 onwards.
- Diet: Altromin Maintenance diet for rats and mice 1324 manufactured by Altromin Spezialfutter GmbH & Co.
- Water: Deep bore-well water passed through reverse osmosis unit was provided in plastic water bottles with stainless steel sipper tubes.
- Acclimation period (Including Oestrus Cycle Evaluation): 18 September 2017 to 06 October 2017.
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.8 to 23.4°C
- Humidity (%): 43 to 69%
- 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: To: 18 September 2017 to 10 December 2017 (till end of last female necropsy) - Route of administration:
- oral: gavage
- Vehicle:
- CMC (carboxymethyl cellulose)
- Remarks:
- 0.5 % w/v Carboxy Methyl Cellulose
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS:
VEHICLE
- Justification for use and choice of vehicle : The test item was in-soluble in distilled water and formed clear homogenous suspension in 0.5 % w/v Carboxy Methyl Cellulose at the concentration of 100 mg/mL (highest dose concentration) as evidenced by the in-house solubility and suspendibility test results. Hence, 0.5 % w/v Carboxy Methyl Cellulose was selected for the test item formulation preparation.
- Concentration in vehicle: G1 (vehicle control): 0 mg/mL; G2 (low dose): 10 mg/mL; G3 (mid dose): 30 mg/mL; G4 (high dose): 100 mg/mL
- Amount of vehicle: 10 mL/kg body weight
- Batch no.: BCBN1690V - Details on mating procedure:
- - M/F ratio per cage: 1:1
- Length of cohabitation: maximum of 14 days.
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy.
- After 14 days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: no
- After successful mating each pregnant female was caged: individually
- Any other deviations from standard protocol: no - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- Homogeneity and dose formulation analysis for dose concentration verification was done by Analytical Chemistry department of Bioneeds India Private Limited.
The analysis was done as per methods detailed in the Study Plan No. BIO-ANM 450. Sampling and analysis of formulations was performed during first week of treatment and during week 4 of the treatment. The samples were collected in duplicates (20 mL each) from top, middle and bottom layer from low and mid dose concentrations, in duplicates (5 mL each) from top, middle and bottom layer from high dose concentrations and from middle layer in duplicate sets from vehicle control.
The collected samples were transferred to Analytical Chemistry department of Bioneeds India Private Limited for dose concentration analysis. One set of aliquot of each formulation was analyzed. The second aliquot was stored as a backup purpose at 2 to 8°C conditions. The second set of samples was discarded, as the analysis results of first set of samples were within the limits. Formulations were considered acceptable, as the mean results were within the range of 85 to 115% of the nominal concentration and the relative standard deviation (% RSD) is ≤10%. - Duration of treatment / exposure:
- Males: two weeks pre-mating, during mating and up to the day before sacrifice during post-mating period (total of 34 days of treatment).
Females: two weeks pre-mating period, during mating, pregnancy (gestation) and up to lactation day 13. - Frequency of treatment:
- Once daily
- Details on study schedule:
- - Age at mating of the mated animals in the study: 14 to 15 weeks
- Dose / conc.:
- 0 mg/kg bw/day
- Remarks:
- vehicle control
- Dose / conc.:
- 100 mg/kg bw/day
- Remarks:
- low dose
- Dose / conc.:
- 300 mg/kg bw/day
- Remarks:
- mid dose
- Dose / conc.:
- 1 000 mg/kg bw/day
- Remarks:
- high dose
- No. of animals per sex per dose:
- 12 animals per sex per dose (group)
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale:
The doses of 0, 100, 300 and 1000 mg/kg body weight for vehicle control, low dose, mid dose and high dose respectively were selected in consultation with the sponsor based on the results of Dose Range Finding Study for Reproduction/Developmental Toxicity Screening Test of Hostacerin DGSB by Oral Gavage in Sprague Dawley Rats (Bioneeds Study No.: BIO-TX 2516).
These doses were selected as the test item Hostacerin DGSB when administered orally to Sprague Dawley males for two weeks prior to mating, during mating and up to the day before sacrifice during post-mating period (total of at 30 days treatment) and females were treated for two weeks pre-mating period, during mating, pregnancy (gestation) and up to lactation day 13 after which the pups were sacrificed on lactation day 13, females (dams) were sacrificed on lactation day 14 did not reveal any treatment related effects at all the tested doses (100, 300 and 1000 mg/kg body weight).
- Rationale for animal assignment:
The animals were weighed and arranged in ascending order of their body weights. These body weight stratified animals were distributed to all the groups using Microsoft Excel Spreadsheet, such that body weight variation of animals selected for the study did not exceed ± 20% (+12.02% and -11.65% for males and +12.09% and -10.47% for females) of the mean body weight of each sex. The grouping was done one day prior to the initiation of treatment. Body weight of the animals was analyzed statistically for mean body weight to rule out the statistical significant difference between groups within each sex. - Positive control:
- not applicable
- Parental animals: Observations and examinations:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: once daily
- Cage side observations checked in table 1 were included.
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly once were included in (table 1)
BODY WEIGHT: Yes
- Time schedule for examinations: first day of dosing, weekly thereafter and at termination. The
females were weighed on gestation days 0, 7, 14 and 20 during pregnancy and on days 1, 4, 7 and 1
3 during lactation period. (table 2, 3, 9, 10, 12 & 13)
FOOD CONSUMPTION: Food consumption for each animal determined and mean daily diet consumption calculated as
g food/kg body weight/day: Yes; weekly once; gestation days 0 to 7, 7 to 14 and 14 to 20 during
pregnancy and day 1 to 4, 4 to 7, 7 to 13 during lactation period. (table 4, 11 & 14) - Oestrous cyclicity (parental animals):
- Two weeks after five days of acclimatization to evaluate its normal oestrus cyclicity (4 to 5 days), daily from the beginning of the treatment period until evidence of mating and on the day of sacrifice for females.
- Sperm parameters (parental animals):
- Parameters examined: testis weight, epididymis weight
Special emphasis on stages of spermatogenesis and histology of interstitial testicular cell structure. - Litter observations:
- STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no
PARAMETERS EXAMINED
The following parameters were examined in offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities, anogenital distance (AGD), presence of nipples/areolae in male pups
GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities - Postmortem examinations (parental animals):
- SACRIFICE
- Male animals: All animals [after completion of mating period (34 days of treatment)]
- Maternal animals: All animals [on lactation day 14]
GROSS NECROPSY
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]
HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table [26 and 27] (i.e. Epidymides, Prostate and Seminal vesicles and coagulating glands, Testes Uterus with cervix Thyroid along with parathyroid) were prepared for microscopic examination and weighed, respectively. - Postmortem examinations (offspring):
- SACRIFICE
- The offspring were sacrificed on lactation day 13.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:
GROSS NECROPSY
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]
HISTOPATHOLOGY / ORGAN WEIGTHS
Thyroid along with parathyroid was collected from one male and one female pup per litter on lactation day 13 and were prepared for microscopic examination - Statistics:
- The data was subjected to various statistical analysis using SPSS software version 22. All analysis and comparisons were evaluated at the 95% level of confidence (P<0.05)
The statistical analysis was followed to the parameters as mentioned below table: Body weight, Change in body weight, Feed consumption, Copulatory interval, Gestation length, Organ
weights, Anogenital distance, Mean pup weight, Pre implantation loss, Pre natal loss, Post natal loss, Post implantation loss:- Parametric - One-way ANOVA with Dunnett’s post test.
Corpora lutea/dam, Implantations/dam, No. of pups/dam, Sex ratio, Litter size, Pup weight:- Non Parametric - Kruskal-Wallis followed by the Mann-Whitney test. Pregnancy rate. Dams with live pups, Dams with dead pups, No. of litters with/without dead pups, No.of litters with/without resorptions:- Cross Tabs - Chi-square test/ Fischer's Exact Test. - Reproductive indices:
- Male Fertility Index (%) = Number of Males Impregnating Females
----------------------------------------------------------- x 100
Number of Males used for Mating
Female Fertility Index (%) = Number of Females confirmed with Pregnancy
------------------------------------------------------------------ x 100
Number of Females used for Mating
Parturition (%) = Number of Parturitions
--------------------------------- x 100
Number of Pregnancies
Copulatory Interval (Days) = Date of Initiation of Cohabitation - Date of Confirmation of Mating
Gestation Length (days) = Date of Delivery - Date of Confirmation of Mating - Offspring viability indices:
- Male/Female Sex Ratio = Number Male Pups Per Dam
------------------------------------------
Number Female Pups Per Dam
Live Birth Index (%) = Number of Viable Pups Born
----------------------------------------- x 100
Number of Pups Born
Lactation Day 4
Pup Survival Index (%) = Number of Viable Pups on Lactation Day 4
---------------------------------------------------------------x 100
Number of Pups Born
Lactation Day 7
Pup Survival Index (%) = Number Of Viable Pups on Lactation Day 7
--------------------------------------------------------------- x 100
Number Of Viable Pups on Lactation Day 4
Lactation Day 13
Pup Survival Index (%) = Number Of Viable Pups on Lactation Day 13
---------------------------------------------------------------- x 100
Number Of Viable Pups on Lactation Day 7 - Clinical signs:
- no effects observed
- Description (incidence and severity):
- There were no clinical signs of toxicity observed at any of the tested dose group animals of either sex during the experimental period.
- Dermal irritation (if dermal study):
- not specified
- Description (incidence and severity):
- not applicable
- Mortality:
- no mortality observed
- Description (incidence):
- There were no mortality/morbidity observed at any of the tested dose group animals of either sex during the experimental period
- Body weight and weight changes:
- effects observed, non-treatment-related
- Description (incidence and severity):
- There were no changes observed in mean body weight and percent change in body weight with respect to day 1 at any of the tested dose group of either sex during the experimental period.
There were no treatment related changes observed in the gestation body weight and percent change in gestation body weight during gestation period at any of the tested dose group animals when compared with vehicle control group animals. However, statistical significant increase in mean body weight on gestation day 14 at G3 group animals was observed when compared with vehicle control group animals. This change is considered as incidental as there were no changes noted during the other gestation periods and there were no effects noted in feed consumption at this period.
There were no treatment related changes observed in the lactation body weight and percent change in lactation body weight at any of the tested dose group animals when compared with vehicle control group animals. - Food consumption and compound intake (if feeding study):
- effects observed, non-treatment-related
- Description (incidence and severity):
- There were no changes observed in feed consumption at any of the tested dose group animals of either sex during the experimental period.
There were no changes observed in the feed consumption during gestation period at any of the tested dose group animals when compared with vehicle control group animals. However, statistical significant increase in mean feed consumption during gestation day 0 to 7 was noted at low and high dose group animals when compared with vehicle control group animals. This change is considered as incidental and not treatment related as there were no effects noted in mean body weight at this duration and the change is inconsistent.
There were no changes observed in the feed consumption during lactation period at any of the tested dose group animals when compared with vehicle control group animals. - Food efficiency:
- not examined
- Description (incidence and severity):
- not applicable
- Water consumption and compound intake (if drinking water study):
- not examined
- Description (incidence and severity):
- not applicable
- Ophthalmological findings:
- not examined
- Description (incidence and severity):
- not applicable
- Haematological findings:
- not examined
- Description (incidence and severity):
- not applicable
- Clinical biochemistry findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- There were no treatment related changes observed in serum T4 levels in males, females (pregnant), lactation day 4 and 13 pups at any of the tested dose groups when compared with vehicle control group.
However, statistically significant decrease in T4 levels at all the tested dose group males when compared with vehicle control group males and statistically significant decrease in T4 levels at mid and high dose group females when compared with vehicle control group females was noted.
This significant variation could be due to two males in G1 group (Animal No. Rc3917 and Rc3922) with a serum T4 level of 139.88 and 134.97 respectively which are significantly more than other animals of same group. In case of females the variation is not dose dependant. Hence, the variations observed in the T4 levels are considered to be incidental. - Urinalysis findings:
- not examined
- Description (incidence and severity):
- not applicable
- Behaviour (functional findings):
- not examined
- Description (incidence and severity):
- not applicable
- Immunological findings:
- not examined
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Description (incidence and severity):
- not applicable
- Histopathological findings: non-neoplastic:
- no effects observed
- Description (incidence and severity):
- No treatment related histopathological findings noticed in ovaries, testes and epididymides (with special emphasis on stages of spermatogenesis and histopathology of interstitial testicular cell structure) of the animals from the high dose group during histopathological examination.
Minimal grade and focally distributed interstitial mononuclear cell [MNC] infiltration, unilateral was observed in epididymis of one male animal of G4 group. This lesion was considered as incidental finding because of lack of consistency and similar finding was observed in concurrent control animal.
Testes, which were screened with special emphasis on stages of spermatogenesis and interstitial testicular cell structure, revealed normal progression of the spermatogenic cycle and presence of all expected associations. - Histopathological findings: neoplastic:
- not examined
- Description (incidence and severity):
- not applicable
- Reproductive function: oestrous cycle:
- no effects observed
- Description (incidence and severity):
- There were no changes observed in the oestrus cyclicity at any of the tested dose group females during pre-mating treatment, mating treatment and on lactation day 14.
- Reproductive function: sperm measures:
- no effects observed
- Description (incidence and severity):
- no effects were noted.
- Reproductive performance:
- no effects observed
- Description (incidence and severity):
- No effects were noted. There were no treatment related changes observed in the copulatory interval and number of conceiving days at any of the tested dose group animals when compared with vehicle control group animals. There were no changes observed in the gestation length, implantation index, pre-implantation loss, post-implantation loss, number of pups delivered, sex ratio and live birth index of each litter at any of the tested dose group animals when compared with vehicle control group animals.
- 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: No adverse effects observed
- Clinical signs:
- no effects observed
- Mortality / viability:
- mortality observed, non-treatment-related
- Description (incidence and severity):
- Mean live birth index is 99.3, 87.85, 98.52 and 100 % for dose groups G1, G2, G3, and G4, respectively. Total number of dead pups at birth were 1, 3, 2 and 0 for dose groups G1, G2, G3, and G4, respectively.
Mean pup survival index (LD 1-4) is 98.48, 100, 100, and 100 % for dose groups G1, G2, G3, and G4, respectively. Total number of dead pups were 2, 0, 0 and 0 for dose groups G1, G2, G3, and G4, respectively. - Body weight and weight changes:
- no effects observed
- Food consumption and compound intake (if feeding study):
- not examined
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- not examined
- Haematological findings:
- not examined
- Clinical biochemistry findings:
- not examined
- Urinalysis findings:
- not examined
- Sexual maturation:
- no effects observed
- Description (incidence and severity):
- There were no changes observed in anogenital distance ratio of both male and female pups, recorded on lactation day 4 at any dose group litters when compared with vehicle control group litters.
There were no occurrences of nipples in male pups of dams of the tested dose group litters when compared with vehicle control group litters. - Organ weight findings including organ / body weight ratios:
- not examined
- Gross pathological findings:
- no effects observed
- Histopathological findings:
- no effects observed
- Other effects:
- no effects observed
- Description (incidence and severity):
- There were no treatment related changes observed in serum T4 levels in lactation day 4 and 13 pups at any of tested dose groups when compared with vehicle control group.
- Behaviour (functional findings):
- not examined
- Developmental immunotoxicity:
- not examined
- Key result
- Dose descriptor:
- NOAEL
- Generation:
- F1
- Effect level:
- >= 1 000 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: No adverse effect s observed
- Key result
- Critical effects observed:
- no
- Key result
- Reproductive effects observed:
- no
- Conclusions:
- Based on the results of the OECD 421 screening study in rats discussed, the No Observed Adverse Effect Level (NOAEL) of the test item Hostacerin DGSB was found to be 1000 mg/kg body weight when administered to the males for two weeks pre-mating, during mating and up to the day before sacrifice during post-mating period (total of 34 days), to the females for two weeks pre-mating, during mating, pregnancy (gestation) and up to lactation day 13 observation under the experimental conditions employed in the study.
- Executive summary:
The test item, Hostacerin DGSB was evaluated for possible adverse effects following repeated oral dosing to males for 34 days, to females for two weeks pre-mating period, during mating period, during pregnancy (gestation) and up to lactation day 13 to evaluate effects of test item on male and female reproductive performance such as gonadal function, mating behaviour, conception, development of the conceptus, parturition, and early neonatal development.
A total of 96 (48 males + 48 females) Sprague Dawley rats were distributed to four groups. Each group (G1, G2, G3and G4) consisted of 12 males and 12 females. The animals in G1 group were administered with vehicle[0.5% w/v Carboxy Methyl Cellulose],animals in G2, G3 and G4groups were administered withtest itemat dose levels of100, 300 and 1000 mg/kg body weight for low dose, mid dose and high dose groups respectively. The vehicle and test item formulations were administered orally by gavage at the dose volume of 10 mL/kg body weight. Males were treated for two weeks pre-mating, during mating and up to the day before sacrifice during post-mating period (total of 34 days of treatment). The females were treated for two weeks pre-mating period, during mating, pregnancy (gestation) and up to lactation day 13 after which the pups were sacrificed on lactation Day 13 and females (dams) were sacrificed on lactation day 14 after overnight fasting (water allowed). Dose formulation analysis for homogeneity and concentration verification was performed during week 1 and 4 of the treatment period and the results are found to be within the acceptable range of 85 to 115 % the nominal concentration and the % RSD is less than 10.0%.All animals were observed for clinical signs once daily, mortality and morbidity twice daily, detailed clinical examination weekly once, body weight and feed consumption weekly once.The serum collected from adult males and females and from lactation day 4 and 13 pups representing each per litter was screened for T4 levels. Females were observed for oestrus cyclicity during pre-mating treatment and mating treatment period and the dams on lactation day 14 prior to sacrifice. The females were observed for copulatory interval and all the adult animals were observed for mating and fertility index. Each litter was examined after delivery (lactation day 1) and the number and sex of pups (litter size), stillbirths (dead pups born on day 1) and live births were recorded. The dams were observed for body weights and feed consumption during gestation and lactation periods, gestation length, live birth index, number of pups, sex ratio and pup survival indexthroughout the lactation period. The pups were observed forclinical signs and once daily from PND 1 to 13. The both male and female pupweights were recorded separately on lactation days 1, 4, 7 and 13.The anogenital distance of each pup was measured on lactation day 4. The numbers of nipples/areolae in male pups were counted on lactation day 13.Gross pathology and organ weighing were performed on day 35 for males. The gross pathology was performed on lactation day 13 for pups, lactation day 14 for dams. The number of corpora lutea and implantation sites for dams were recorded during necropsy.
All animals did not reveal any clinical signs of toxicity, no mortality or morbidity observed throughout the experimental period. No treatment related changes in body weight, percent change in body weight, feed consumption, organ weights (both absolute and relative) were observed in both the sex of all tested dose group animals. There were no treatment related changes observed in serum T4 levels of adult animals and in serum T4 levels of pups at any of the tested dose groups.
Females did not reveal any treatment related changes in oestrus cyclicity, copulatory interval, fertility index, body weights and feed consumption during gestation and lactation periods, gestation length, implantation index, pre-implantation loss, post-impalntation loss, number of pups delivered, live birth index of each litter, sex ratio and pup survival index at all the tested dose groups throughout the lactation periodwhen compared with vehicle control group animals.
All pups did not reveal any clinical signs or external anomalies throughout the lactation period. No treatment related changes in pup weights, anogenital distance ratio were noted. No occurrences of nipples in male pups at any of the tested dose groups and vehicle control group.
All animals of all dose groups did not reveal any treatment related gross pathological and histopathological findings compared to vehicle control group.
Referenceopen allclose all
TABLE 1. SUMMARY OF CLINICAL SIGNS OF TOXICITY AND MORTALITY RECORD
Group, Sex & Dose (mg/kg body weight) |
No. of Animals |
Clinical Signs of Toxicity |
Mortality (No. of Mortality/ No. of Animals dosed) |
G1, M & 0 |
6 |
N |
0/6 |
G2, M & 100 |
6 |
N |
0/6 |
G3, M & 300 |
6 |
N |
0/6 |
G4, M & 1000 |
6 |
N |
0/6 |
M: Male; N: Normal
TABLE 1 (Contd..,). SUMMARYOF CLINICAL SIGNS OF TOXICITYAND MORTALITY RECORD
Group, Sex & Dose (mg/kg body weight) |
No. of Animals |
Clinical Signs of Toxicity |
Mortality (No. of Mortality/ No. of Animals dosed) |
G1, F & 0 |
6 |
N |
0/6 |
G2, F & 100 |
6 |
N |
0/6 |
G3, F & 300 |
6 |
N |
0/6 |
G4, F & 1000 |
6 |
N |
0/6 |
F: Female; N: Normal
TABLE 2. SUMMARY OF BODY WEIGHTS (g) RECORD
Group, Sex & Dose (mg/kg body weight) |
|
Body Weight (g) on Days |
|||||
1 |
8 |
15 |
22 |
29 |
|||
G1, M & 0 |
Mean |
331.25 |
362.71 |
386.31 |
409.37 |
421.80 |
|
±SD |
13.90 |
10.59 |
7.59 |
9.44 |
8.63 |
||
n |
6 |
6 |
6 |
6 |
6 |
||
G2, M & 100 |
Mean |
331.77 |
357.98 |
381.55 |
405.92 |
418.53 |
|
±SD |
14.34 |
10.81 |
10.70 |
10.11 |
8.89 |
||
n |
6 |
6 |
6 |
6 |
6 |
||
G3, M & 300 |
Mean |
332.10 |
361.95 |
380.97 |
402.04 |
415.77 |
|
±SD |
16.49 |
9.19 |
8.89 |
7.83 |
6.15 |
||
n |
6 |
6 |
6 |
6 |
6 |
||
G4, M & 1000 |
Mean |
334.17 |
362.38 |
382.72 |
406.87 |
422.84 |
|
±SD |
16.95 |
13.77 |
10.38 |
10.84 |
12.60 |
||
n |
6 |
6 |
6 |
6 |
6 |
M: Male; SD: Standard Deviation; n: Number of Animals
TABLE 2 (Contd..,). SUMMARY OF BODY WEIGHTS (g) RECORD
Group, Sex & Dose (mg/kg body weight) |
|
Body Weight (g) on Days |
||||
1 |
8 |
15 |
22# |
|||
G1, F & 0 |
Mean |
240.69 |
252.13 |
260.78 |
267.33 |
|
±SD |
8.35 |
7.06 |
2.96 |
- |
||
n |
6 |
6 |
6 |
1 |
||
G2, F & 100 |
Mean |
241.04 |
253.47 |
263.23 |
267.77 |
|
±SD |
7.44 |
6.97 |
7.49 |
2.46 |
||
n |
6 |
6 |
6 |
2 |
||
G3, F & 300 |
Mean |
240.83 |
253.07 |
262.61 |
269.33 |
|
±SD |
7.53 |
7.63 |
7.60 |
- |
||
n |
6 |
6 |
6 |
1 |
||
G4, F & 1000 |
Mean |
242.16 |
255.14 |
264.16 |
260.79 |
|
±SD |
7.12 |
6.55 |
6.73 |
- |
||
n |
6 |
6 |
6 |
1 |
F: Female; SD: Standard Deviation; n: Number of Animals; -: Not applicable
#:The body weight of females only in cohabitation was consideredfor mean calculation.
TABLE 3. SUMMARY OF PERCENTCHANGE IN BODY WEIGHT (%) WITH RESPECT TO DAY 1 RECORD
Group, Sex & Dose (mg/kg body weight) |
|
Percent Change in Body Weight (%) during Days |
||||
1 to 8 |
1 to 14 |
1 to 21 |
1 to 29 |
|||
G1, M & 0 |
Mean |
9.60 |
16.74 |
23.68 |
27.44 |
|
±SD |
4.14 |
3.88 |
3.24 |
3.12 |
||
n |
6 |
6 |
6 |
6 |
||
G2, M & 100 |
Mean |
7.99 |
15.13 |
22.55 |
26.34 |
|
±SD |
3.53 |
4.71 |
6.33 |
6.00 |
||
n |
6 |
6 |
6 |
6 |
||
G3, M & 300 |
Mean |
9.11 |
14.85 |
21.22 |
25.38 |
|
±SD |
3.05 |
3.60 |
3.81 |
4.49 |
||
n |
6 |
6 |
6 |
6 |
||
G4, M & 1000 |
Mean |
8.51 |
14.65 |
21.88 |
26.65 |
|
±SD |
2.00 |
2.90 |
2.92 |
2.94 |
||
n |
6 |
6 |
6 |
6 |
M: Male; SD: Standard Deviation; n: Number of Animals
TABLE 3 (Contd..,). SUMMARY OF CHANGE IN BODY WEIGHT (%) WITH RESPECT TO DAY 1 RECORD
Group, Sex & Dose (mg/kg body weight) |
Percent Change in Body Weight (%) during Days |
||||
1 to 8 |
1 to 14 |
1 to 21# |
|||
G1, F & 0 |
Mean |
4.78 |
8.45 |
8.96 |
|
±SD |
1.17 |
3.52 |
- |
||
n |
6 |
6 |
1 |
||
G2, F & 100 |
Mean |
5.17 |
9.22 |
13.13 |
|
±SD |
1.05 |
1.19 |
1.35 |
||
n |
6 |
6 |
2 |
||
G3, F & 300 |
Mean |
5.09 |
9.06 |
15.52 |
|
±SD |
1.11 |
1.68 |
- |
||
n |
6 |
6 |
1 |
||
G4, F & 1000 |
Mean |
5.37 |
9.10 |
11.71 |
|
±SD |
0.70 |
1.33 |
- |
||
n |
6 |
6 |
1 |
F: Female; SD: Standard Deviation; n: Number of Animals; -: Not applicable
#:The percent change in body weight of females only in cohabitation was consideredfor mean calculation.
TABLE 4. SUMMARY OF FEED CONSUMPTION (g/rat/day) RECORD
Group, Sex & Dose |
Week 1 Feed Consumption (Day 1 to 8) |
Week 2 Feed Consumption (Day 8 to 14) |
Post mating Feed Consumption (Day 25 to 29) |
Post mating Feed Consumption (Day 29 to 30) |
||
G1, M & 0 |
Mean |
24.38 |
27.37 |
28.83 |
28.18 |
|
±SD |
1.10 |
0.71 |
0.63 |
1.16 |
||
n |
6 |
6 |
6 |
6 |
||
G2, M & 100 |
Mean |
25.20 |
28.29* |
29.72* |
28.07 |
|
±SD |
0.49 |
0.47 |
0.57 |
0.71 |
||
n |
6 |
6 |
6 |
6 |
||
G3, M & 300 |
Mean |
25.19 |
28.42* |
29.63 |
29.00 |
|
±SD |
0.73 |
0.58 |
0.82 |
0.53 |
||
n |
6 |
6 |
6 |
6 |
||
G4, M & 1000 |
Mean |
25.07 |
28.00 |
29.08 |
28.00 |
|
±SD |
0.72 |
0.60 |
0.71 |
1.13 |
||
n |
6 |
6 |
6 |
6 |
M: Male;SD: Standard Deviation; n: Number ofanimals
*: Statistically significant than the control group (p<0.05)
TABLE 4 (Contd..,). SUMMARY OF FFED CONSUMPTION (g/rat/day) RECORD
Group, Sex & Dose |
Feed Consumption (g) during Premating |
|||
Week 1 |
Week 2 |
|||
G1, F & 0 |
Mean |
19.97 |
21.26 |
|
±SD |
0.55 |
0.53 |
||
n |
6 |
6 |
||
G1, F & 100 |
Mean |
19.86 |
21.35 |
|
±SD |
0.64 |
0.53 |
||
n |
6 |
6 |
||
G3, F & 300 |
Mean |
20.38 |
21.91 |
|
±SD |
0.62 |
0.41 |
||
n |
6 |
6 |
||
G4, F & 1000 |
Mean |
21.81* |
23.19* |
|
±SD |
1.68 |
1.30 |
||
n |
6 |
6 |
F: Female; SD: Standard Deviation;Number of animals
*: Statistically significant than the control group (p<0.05)
TABLE 5. SUMMARY OF ABSOLUTE ORGAN WEIGHT (g) RECORD
Group, Sex & Dose (mg/kg body weight) |
Epididymis |
Testes |
Prostate + Seminal vesicles with coagulating glands (PSC) |
|
G1, M & 0 |
Mean |
1.7403 |
3.5235 |
3.8015 |
±SD |
0.0339 |
0.2507 |
0.4641 |
|
n |
6 |
6 |
6 |
|
G2, M & 100 |
Mean |
1.5891 |
3.4216 |
3.4161 |
±SD |
0.0593 |
0.2754 |
0.2124 |
|
n |
6 |
6 |
6 |
|
G3, M & 300 |
Mean |
1.6754 |
3.4561 |
3.5618 |
±SD |
0.1192 |
0.2128 |
0.3601 |
|
n |
6 |
6 |
6 |
|
G4, M & 1000 |
Mean |
1.6969 |
3.4100 |
3.7384 |
±SD |
0.1082 |
0.2617 |
0.1575 |
|
n |
6 |
6 |
6 |
M: Male;SD: Standard Deviation; n: Number of animals
TABLE 5 (Contd..,). SUMMARY OF ABSOLUTE ORGAN WEIGHT (g) RECORD
Group, Sex & Dose (mg/kg body weight) |
Uterus |
Ovaries |
G1, F & 0 |
0.4807 |
0.1691 |
0.0440 |
0.0339 |
|
6 |
6 |
|
G2, F & 100 |
0.5102 |
0.1695 |
0.0627 |
0.0335 |
|
6 |
6 |
|
G3, F & 300 |
0.4604 |
0.1868 |
0.0367 |
0.0255 |
|
6 |
6 |
|
G4, F & 1000 |
0.4749 |
0.1799 |
0.0424 |
0.0206 |
|
6 |
6 |
F: Female;SD: Standard Deviation; n: Number of animals
TABLE 6. SUMMARY OF TERMINAL BODY WEIGHT (g) AND RELATIVE ORGAN WEIGHT (%) RECORD
Group, Sex & Dose (mg/kg body weight) |
Fasting Body Weight (g) |
Epididymis |
Testes |
Prostate + Seminal vesicles with coagulating glands (PSC) |
|
G1, M & 0 |
Mean |
402.18 |
0.4329 |
0.8762 |
0.9471 |
±SD |
8.40 |
0.0153 |
0.0609 |
0.1324 |
|
n |
6 |
6 |
6 |
6 |
|
G2, M & 100 |
Mean |
397.70 |
0.3996 |
0.8602 |
0.8591 |
±SD |
5.44 |
0.0136 |
0.0650 |
0.0537 |
|
n |
6 |
6 |
6 |
6 |
|
G3, M & 300 |
Mean |
394.75 |
0.4243 |
0.8753 |
0.9027 |
±SD |
6.35 |
0.0273 |
0.0483 |
0.0944 |
|
n |
6 |
6 |
6 |
6 |
|
G4, M & 1000 |
Mean |
400.77 |
0.4233 |
0.8522 |
0.9331 |
±SD |
12.55 |
0.0220 |
0.0797 |
0.0373 |
|
n |
6 |
6 |
6 |
6 |
M: Male;SD: Standard Deviation; n: Number of animals
TABLE 6 (Contd..,). SUMMARY OF RELATIVE ORGAN WEIGHT (%) RECORD
Group, Sex & Dose (mg/kg body weight) |
Fasting Body Weight (g) |
Uterus |
Ovaries |
|
G1, F & 0 |
Mean |
283.56 |
0.1693 |
0.0596 |
±SD |
12.27 |
0.0106 |
0.0117 |
|
n |
6 |
6 |
6 |
|
G2, F & 100 |
Mean |
298.89 |
0.1722 |
0.0572 |
±SD |
29.01 |
0.0277 |
0.0127 |
|
n |
6 |
6 |
6 |
|
G3, F & 300 |
Mean |
279.78 |
0.1653 |
0.0668 |
±SD |
13.38 |
0.0197 |
0.0093 |
|
n |
6 |
6 |
6 |
|
G4, F & 1000 |
Mean |
278.56 |
0.1716 |
0.0650 |
±SD |
23.11 |
0.0223 |
0.0096 |
|
n |
6 |
6 |
6 |
F: Female;SD: Standard Deviation; n: Number of animals
TABLE 7. SUMMARY OF VAGINAL SMEAR EXAMINATION RECORD FOR DETERMINATION OF OESTRUS CYCLICITY
Group & Dose |
No. of Females |
No. of Females with Regular Oestrus Cyclicity during Pre-mating, mating and on Lactation day 14 |
No. of Females with Irregular Oestrus Cyclicity during Pre-mating, mating and on Lactation day 14 |
G1 & 0 |
6 |
6 |
0 |
G2 & 100 |
6 |
6 |
0 |
G3 & 300 |
6 |
6 |
0 |
G4 & 1000 |
6 |
6 |
0 |
TABLE 8. SUMMARY OF COHABITATION RECORD
Group & Dose |
Copulatory Interval (Days) |
|
Conceiving Days (1 to 5) |
Conceiving Days (6 to 20) |
|
G1 & 0 |
Mean |
6.50 |
n |
2 |
4 |
±SD |
3.33 |
||||
n |
6 |
% |
33.3 |
66.7 |
|
G2 & 100 |
Mean |
5.17 |
n |
4 |
2 |
±SD |
3.97 |
||||
n |
6 |
% |
66.7 |
33.3 |
|
G3 & 300 |
Mean |
4.00 |
n |
4 |
2 |
±SD |
3.41 |
||||
n |
6 |
% |
66.7 |
33.3 |
|
G4 & 1000 |
Mean |
5.33 |
n |
4 |
2 |
±SD |
2.80 |
||||
n |
6 |
% |
66.7 |
33.3 |
SD: Standard Deviation; n: Number of animals; %: Percent
TABLE 9. SUMMARY OF BODY WEIGHT (g) RECORD DURING GESTATION
Group & Dose |
Body weight (g) on Gestation Days |
||||
0 |
7 |
14 |
20 |
||
G1 & 0 |
Mean |
265.73 |
277.18 |
312.04 |
378.43 |
±SD |
3.75 |
6.04 |
16.21 |
20.04 |
|
n |
6 |
6 |
6 |
6 |
|
G2 & 100 |
Mean |
268.66 |
281.41 |
328.45 |
398.77 |
±SD |
10.15 |
10.55 |
11.72 |
22.33 |
|
n |
6 |
6 |
6 |
6 |
|
G3 & 300 |
Mean |
264.01 |
275.66 |
311.19 |
380.17 |
±SD |
7.22 |
3.86 |
5.59 |
25.44 |
|
n |
6 |
6 |
6 |
6 |
|
G4 & 1000 |
Mean |
266.89 |
278.45 |
313.20 |
377.31 |
±SD |
5.22 |
5.19 |
10.25 |
24.42 |
|
n |
6 |
6 |
6 |
6 |
SD: Standard Deviation; n: Number of animals
TABLE 10. SUMMARY OF CHANGE IN BODY WEIGHT (%) RECORD DURING GESTATION
Group & Dose |
Change in body weight (%) during Gestation |
||||
0 to 7 |
7 to 14 |
14 to 20 |
0 to 20 |
||
G1 & 0 |
Mean |
4.30 |
12.53 |
21.31 |
42.39 |
±SD |
0.98 |
3.97 |
3.40 |
6.73 |
|
n |
6 |
6 |
6 |
6 |
|
G2 & 100 |
Mean |
4.77 |
16.79 |
21.36 |
48.65 |
±SD |
2.17 |
4.20 |
3.91 |
10.80 |
|
n |
6 |
6 |
6 |
6 |
|
G3 & 300 |
Mean |
4.45 |
12.91 |
22.10 |
44.25 |
±SD |
1.90 |
2.89 |
6.40 |
12.93 |
|
n |
6 |
6 |
6 |
6 |
|
G4 & 1000 |
Mean |
4.34 |
12.50 |
20.41 |
41.45 |
±SD |
1.57 |
3.71 |
5.16 |
10.04 |
|
n |
6 |
6 |
6 |
6 |
SD: Standard Deviation; n: Number of animals
TABLE 11. SUMMARY OF FEED CONSUMPTION (g/dam/day) RECORD DURING GESTATION
Group & Dose |
Feed Consumption (g) during Gestation |
||||
GD 0 to 7 |
GD 7 to 14 |
GD 14 to 20 |
|||
G1 & 0 |
Mean |
22.25 |
25.64 |
27.24 |
|
±SD |
1.53 |
1.41 |
1.90 |
||
n |
6 |
6 |
6 |
||
G2 & 100 |
Mean |
22.60 |
25.44 |
25.49 |
|
±SD |
1.14 |
1.93 |
2.44 |
||
n |
6 |
6 |
6 |
||
G3 & 300 |
Mean |
22.73 |
24.57 |
25.98 |
|
±SD |
0.60 |
2.00 |
3.02 |
||
n |
6 |
6 |
6 |
||
G4 & 1000 |
Mean |
22.33 |
26.08 |
25.96 |
|
±SD |
1.41 |
2.16 |
0.89 |
||
n |
6 |
6 |
6 |
SD: Standard Deviation; n: Number of animals
TABLE 12. SUMMARY OF BODY WEIGHT (g) RECORD DURING LACTATION
Group & Dose |
Body Weight (g) on Lactation Day |
||||
1 |
4 |
7 |
13 |
||
G1 & 0 |
Mean |
279.76 |
287.68 |
294.42 |
304.97 |
±SD |
19.67 |
21.31 |
18.89 |
15.00 |
|
n |
6 |
6 |
6 |
6 |
|
G2 & 100 |
Mean |
292.41 |
299.66 |
307.01 |
320.33 |
±SD |
34.67 |
35.09 |
34.00 |
30.14 |
|
n |
6 |
6 |
6 |
6 |
|
G3 & 300 |
Mean |
276.63 |
283.55 |
290.71 |
301.02 |
±SD |
16.51 |
16.40 |
16.14 |
14.71 |
|
n |
6 |
6 |
6 |
6 |
|
G4 & 1000 |
Mean |
273.45 |
279.79 |
286.38 |
296.82 |
±SD |
23.69 |
23.94 |
24.64 |
24.15 |
|
n |
6 |
6 |
6 |
6 |
SD: Standard Deviation; n: Number of animals
TABLE 13. SUMMARY OF CHANGE IN BODY WEIGHT (%) RECORD DURING LACTATION
Group & Dose |
Change in body weight (%) during Lactation |
|||
1 to 4 |
4 to 7 |
7 to 13 |
||
G1 & 0 |
Mean |
2.82 |
2.41 |
3.68 |
±SD |
1.67 |
1.12 |
2.21 |
|
n |
6 |
6 |
6 |
|
G2 & 100 |
Mean |
2.49 |
2.52 |
4.52 |
±SD |
1.15 |
1.02 |
2.74 |
|
n |
6 |
6 |
6 |
|
G3 & 300 |
Mean |
2.51 |
2.54 |
3.59 |
±SD |
0.57 |
0.66 |
1.59 |
|
n |
6 |
6 |
6 |
|
G4 & 1000 |
Mean |
2.33 |
2.35 |
3.69 |
±SD |
1.13 |
0.59 |
1.51 |
|
n |
6 |
6 |
6 |
SD: Standard Deviation; n: Number of animals
TABLE 14. SUMMARY OF FEED CONSUMTION (g/dam/day) RECORD DURING LACTATION
Group & Dose |
Feed Consumption (g) during Lactation |
||||
LD 1 to 4 |
LD 4 to 7 |
LD 7 to 14 |
|||
G1 & 0 |
Mean |
32.09 |
40.27 |
40.91 |
|
±SD |
2.46 |
3.65 |
6.25 |
||
n |
6 |
6 |
6 |
||
G2 & 100 |
Mean |
31.75 |
40.42 |
43.24 |
|
±SD |
3.79 |
5.40 |
5.29 |
||
n |
6 |
6 |
6 |
||
G3 & 300 |
Mean |
29.77 |
39.58 |
44.18 |
|
±SD |
3.34 |
4.87 |
5.27 |
||
n |
6 |
6 |
6 |
||
G4 & 1000 |
Mean |
32.98 |
45.05 |
41.47 |
|
±SD |
5.93 |
6.65 |
6.34 |
||
n |
6 |
6 |
6 |
SD: Standard Deviation; n: Number of animals; LD: Lactation Day
TABLE 15. SUMMARY OF GESTATION LENGTH AND DELIVERY RECORD
Group & Dose |
|
Gestation Length (Days) |
|
Delivery Record at Birth |
|||||||||
Litter Size (No.) |
Live Pups (No.) |
Dead Pups (No.) |
Sex Ratio (M/F) at Birth |
Live Birth Index |
|||||||||
Total (No.) |
Male (No.) |
Female (No.) |
Total (No.) |
Male (No.) |
Female (No.) |
||||||||
G1 & 0 |
Mean |
22.17 |
12.33 |
12.33 |
6.50 |
5.83 |
0.00 |
0.00 |
0.00 |
1.30 |
100.00 |
||
±SD |
0.41 |
2.94 |
2.94 |
0.84 |
2.56 |
0.00 |
0.00 |
0.00 |
0.59 |
0.00 |
|||
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
|||
G2 & 100 |
Mean |
21.83 |
11.50 |
11.33 |
4.67* |
6.67 |
0.17 |
0.00 |
0.17 |
0.80* |
98.33 |
||
±SD |
0.41 |
1.22 |
1.51 |
1.75 |
1.75 |
0.41 |
0.00 |
0.41 |
0.60 |
4.08 |
|||
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
|||
G3 & 300 |
Mean |
22.17 |
12.17 |
11.50 |
3.83* |
7.67 |
0.67 |
0.33 |
0.33 |
0.53* |
91.82 |
||
±SD |
0.41 |
3.54 |
4.32 |
1.47 |
3.27 |
1.21 |
0.82 |
0.52 |
0.25 |
17.17 |
|||
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
|||
G4 & 1000 |
Mean |
22.33 |
12.17 |
12.00 |
4.83* |
7.17 |
0.17 |
0.00 |
0.17 |
0.78* |
98.72 |
||
±SD |
1.03 |
2.32 |
2.28 |
0.75 |
2.23 |
0.41 |
0.00 |
0.41 |
0.45 |
3.14 |
|||
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
M: Male: F: Female;SD: Standard Deviation; n: Number of animals; LD: Lactation Day
*: Statistically significant than the control group (p<0.05)
TABLE 16. SUMMARY OF LITTER OBSERVATION DATA DURING LACTATION
Group & Dose |
|
During LD 1 to 4 |
|||||||||
No. of Pups |
Sex Ratio (M/F) at LD 4 |
No. of Survived Pups during LD 1-4 |
Pup Survival Index |
||||||||
Live Pups (No.) |
|
Dead Pups (No.) |
|||||||||
Total (No.) |
Male (No.) |
Female (No.) |
Total (No.) |
Male (No.) |
Female (No.) |
||||||
G1 & 0 |
Mean |
12.33 |
6.50 |
5.83 |
0.00 |
0.00 |
0.00 |
1.30 |
12.33 |
100.00 |
|
±SD |
2.94 |
0.84 |
2.56 |
0.00 |
0.00 |
0.00 |
0.59 |
2.94 |
0.00 |
||
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
||
G2 & 100 |
Mean |
11.33 |
4.67* |
6.67 |
0.00 |
0.00 |
0.00 |
0.80* |
11.33 |
100.00 |
|
±SD |
1.51 |
1.75 |
1.75 |
0.00 |
0.00 |
0.00 |
0.60 |
1.51 |
0.00 |
||
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
||
G3 & 300 |
Mean |
11.50 |
3.83* |
7.67 |
0.00 |
0.00 |
0.00 |
0.53* |
11.50 |
100.00 |
|
±SD |
4.32 |
1.47 |
3.27 |
0.00 |
0.00 |
0.00 |
0.25 |
4.32 |
0.00 |
||
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
||
G4 & 1000 |
Mean |
12.00 |
4.83* |
7.17 |
0.00 |
0.00 |
0.00 |
0.78* |
12.00 |
100.00 |
|
±SD |
2.28 |
0.75 |
2.23 |
0.00 |
0.00 |
0.00 |
0.45 |
2.28 |
0.00 |
||
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
M: Male: F: Female;SD: Standard Deviation; n: Number of animals; LD: Lactation Day
*: Statistically significant than the control group (p<0.05)
TABLE 16 (Contd..,). SUMMARY OF LITTER OBSERVATION DATA DURING LACTATION
Group & Dose |
|
During LD 4 to 7 |
|||||||||
No. of Pups |
Sex Ratio (M/F) at LD 7 |
No. of Survived Pups during LD 4-7 |
Pup Survival Index LD 4-7 |
||||||||
Live Pups (No.) |
Dead Pups (No.) |
||||||||||
Total (No.) |
Male (No.) |
Female (No.) |
Total (No.) |
Male (No.) |
Female (No.) |
||||||
G1 & 0 |
Mean |
12.33 |
6.50 |
5.83 |
0.00 |
0.00 |
0.00 |
1.30 |
12.33 |
100.00 |
|
±SD |
2.94 |
0.84 |
2.56 |
0.00 |
0.00 |
0.00 |
0.59 |
2.94 |
0.00 |
||
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
||
G2 & 100 |
Mean |
11.33 |
4.67* |
6.67 |
0.00 |
0.00 |
0.00 |
0.80* |
11.33 |
100.00 |
|
±SD |
1.51 |
1.75 |
1.75 |
0.00 |
0.00 |
0.00 |
0.60 |
1.51 |
0.00 |
||
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
||
G3 & 300 |
Mean |
11.50 |
3.83* |
7.67 |
0.00 |
0.00 |
0.00 |
0.53* |
11.50 |
100.00 |
|
±SD |
4.32 |
1.47 |
3.27 |
0.00 |
0.00 |
0.00 |
0.25 |
4.32 |
0.00 |
||
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
||
G4 & 1000 |
Mean |
12.00 |
4.83* |
7.17 |
0.00 |
0.00 |
0.00 |
0.78* |
12.00 |
100.00 |
|
±SD |
2.28 |
0.75 |
2.23 |
0.00 |
0.00 |
0.00 |
0.45 |
2.28 |
0.00 |
||
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
M: Male: F: Female;SD: Standard Deviation; n: Number of animals; LD: Lactation Day
*: Statistically significant than the control group (p<0.05)
TABLE 16 (Contd..,). SUMMARY OF LITTER OBSERVATION DATA DURING LACTATION
Group & Dose |
|
During LD 7 to 14 |
|||||||||
No. of Pups |
Sex Ratio (M/F) at LD 13 |
No. of Survived Pups during LD 7-13 |
Pup Survival Index LD 7-13 |
||||||||
Live Pups (No.) |
Dead Pups (No.) |
||||||||||
Total (No.) |
Male (No.) |
Female (No.) |
Total (No.) |
Male (No.) |
Female (No.) |
||||||
G1 & 0 |
Mean |
12.33 |
6.50 |
5.83 |
0.00 |
0.00 |
0.00 |
1.30 |
12.33 |
100.00 |
|
±SD |
2.94 |
0.84 |
2.56 |
0.00 |
0.00 |
0.00 |
0.59 |
2.94 |
0.00 |
||
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
||
G2 & 100 |
Mean |
11.33 |
4.67* |
6.67 |
0.00 |
0.00 |
0.00 |
0.80* |
11.33 |
100.00 |
|
±SD |
1.51 |
1.75 |
1.75 |
0.00 |
0.00 |
0.00 |
0.60 |
1.51 |
0.00 |
||
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
||
G3 & 300 |
Mean |
11.50 |
3.83* |
7.67 |
0.00 |
0.00 |
0.00 |
0.53* |
11.50 |
100.00 |
|
±SD |
4.32 |
1.47 |
3.27 |
0.00 |
0.00 |
0.00 |
0.25 |
4.32 |
0.00 |
||
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
||
G4 & 1000 |
Mean |
12.00 |
4.83* |
7.17 |
0.00 |
0.00 |
0.00 |
0.78* |
12.00 |
100.00 |
|
±SD |
2.28 |
0.75 |
2.23 |
0.00 |
0.00 |
0.00 |
0.45 |
2.28 |
0.00 |
||
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
M: Male: F: Female;SD: Standard Deviation; n: Number of animals; LD: Lactation Day
*: Statistically significant than the control group (p<0.05)
TABLE 17. SUMMARY OF UTERI OBSERVATION RECORD
Group & Dose |
No. of Corpora luetea |
No. of Implantations |
Implantation Index (%) |
Pre-Implantation Loss (%) |
Post-Implantation Loss (%) |
Pre-natal Loss (No.) |
Post-natal Loss (At birth to LD 13) (%) |
Post-natal Loss (At birth to LD 13) (No.) |
No. of Early Resorptions (No.) |
No. of Late Resorptions (No.) |
|
G1 & 0 |
Mean |
12.83 |
12.33 |
95.69 |
4.31 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
±SD |
2.64 |
2.94 |
4.75 |
4.75 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
|
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
|
G2 & 100 |
Mean |
11.83 |
11.50 |
96.97 |
3.03 |
1.67 |
0.17 |
0.00 |
0.00 |
0.00 |
0.00 |
±SD |
0.75 |
1.22 |
4.69 |
4.69 |
4.08 |
0.41 |
0.00 |
0.00 |
0.00 |
0.00 |
|
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
|
G3 & 300 |
Mean |
12.67 |
12.17 |
96.52 |
3.48 |
8.18 |
0.67 |
0.00 |
0.00 |
0.00 |
0.00 |
±SD |
3.88 |
3.54 |
3.98 |
3.98 |
17.17 |
1.21 |
0.00 |
0.00 |
0.00 |
0.00 |
|
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
|
G4 & 1000 |
Mean |
12.50 |
12.17 |
97.53 |
2.47 |
1.28 |
0.17 |
0.00 |
0.00 |
0.00 |
0.00 |
±SD |
2.43 |
2.32 |
3.83 |
3.83 |
3.14 |
0.41 |
0.00 |
0.00 |
0.00 |
0.00 |
|
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
SD: Standard Deviation; n: Number of animals; LD: Lactation Day
TABLE 18. SUMMARY OF PUP OBSERVATION RECORD DURING LACTATION
Group & Dose |
Pup Observation on LD 1 |
Pup Observation on LD 4 |
Pup Observation on LD 7 |
Pup Observation on LD 13 |
G1 & 0 |
N |
N |
N |
N |
G2 & 100 |
N |
N |
N |
N |
G3 & 300 |
N |
N |
N |
N |
G4 & 1000 |
N |
N |
N |
N |
N: Normal
TABLE 19. SUMMARY OF PUP WEIGHT (g) RECORD
Group & Dose |
Pup Weight (g) on Lactation Day 1 |
Pup Weight (g) on Lactation Day 4 |
Pup Weight (g) on Lactation Day 7 |
Pup Weight (g) on Lactation Day 14 |
||||||
Male |
Female |
Male |
Female |
Male |
Female |
Male |
Female |
|||
G1 & 0 |
Mean |
6.68 |
6.24 |
9.84 |
8.55 |
15.22 |
11.90 |
26.83 |
20.61 |
|
±SD |
0.50 |
0.17 |
0.53 |
0.33 |
0.49 |
0.43 |
0.70 |
0.61 |
||
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
||
G2 & 100 |
Mean |
6.86 |
6.15 |
10.27 |
8.81 |
15.40 |
12.33 |
27.18 |
19.22 |
|
±SD |
0.40 |
0.41 |
0.98 |
0.45 |
0.72 |
0.84 |
1.08 |
4.12 |
||
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
||
G3 & 300 |
Mean |
7.22 |
6.16 |
9.67 |
7.19 |
15.43 |
12.36 |
27.40 |
21.55 |
|
±SD |
0.61 |
0.29 |
1.58 |
3.52 |
0.26 |
0.57 |
1.19 |
0.68 |
||
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
||
G4 & 1000 |
Mean |
7.01 |
6.43 |
10.51 |
9.07 |
15.19 |
12.34 |
26.48 |
21.09 |
|
±SD |
0.75 |
0.41 |
0.51 |
0.46 |
0.57 |
0.37 |
0.50 |
0.67 |
||
n |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
SD: Standard Deviation; n: Number of dams
TABLE 20. SUMMARY OF GROSS PATHOLOGY FINDINGS
Organs/ Lesions |
Group |
G1 |
G2 |
G3 |
G4 |
||||
Dose (mg/kg body weight) |
0 |
100 |
300 |
1000 |
|||||
Sex |
M |
F |
M |
F |
M |
F |
M |
F |
|
No. of Animals |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
|
No Abnormality Detected |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
6 |
M: Male; F: Female
TABLE 20 (Contd..,). SUMMARY OF GROSS PATHOLOGY FINDINGS
Organs/ Lesions |
Group |
G1 |
G2 |
G3 |
G4 |
||||
Dose (mg/kg body weight) |
0 |
100 |
300 |
1000 |
|||||
Sex |
M |
F |
M |
F |
M |
F |
M |
F |
|
No. of Dead pups (At birth) |
0 |
0 |
0 |
1 |
2 |
2 |
0 |
1 |
|
No Abnormality Detected |
0 |
0 |
0 |
1 |
2 |
2 |
0 |
1 |
|
|
No. of pups sacrificed on Lactation Day 13 |
39 |
35 |
28 |
40 |
23 |
46 |
29 |
43 |
No Abnormality Detected |
39 |
35 |
28 |
40 |
23 |
46 |
29 |
43 |
Note:. M: Male; F: Female
TABLE 1. SUMMARYOF CLINICAL SIGNSOF TOXICITY, DETAILED CLINICAL EXAMINATIONAND MORTALITY RECORD
Group, Sex & Dose (mg/kg body weight/day) |
No. of Animals |
Clinical Signs of Toxicity/ Detailed Clinical Examination |
Mortality (No. of Mortality/ No. of Animals dosed) |
G1, M & 0 |
12 |
N |
0/12 |
G2, M & 100 |
12 |
N |
0/12 |
G3, M & 300 |
12 |
N |
0/12 |
G4, M & 1000 |
12 |
N |
0/12 |
M: Male; N: Normal
TABLE 1 (Contd..,). SUMMARYOF CLINICAL SIGNSOF TOXICITY, DETAILED CLINICAL EXAMINATIONAND
MORTALITY RECORD
Group, Sex & Dose (mg/kg body weight/day) |
No. of Animals |
Clinical Signs of Toxicity/ Detailed Clinical Examination |
Mortality (No. of Mortality/ No. of Animals dosed) |
||||
G1, F & 0 |
12 |
N |
0/12 |
||||
G2, F & 100 |
12 |
N |
0/12 |
||||
G3, F & 300 |
12 |
N |
0/12 |
||||
G4, F & 1000 |
12 |
N |
0/12 |
F: Female; N: Normal
TABLE 2. SUMMARY OF BODY WEIGHT (g)RECORD
Group, Sex & Dose (mg/kg body weight/day) |
Body Weight (g) on Days |
|||||||
1 |
7 |
14 |
21 |
28 |
34 |
|||
G1, M & 0 |
Mean |
331.48 |
355.57 |
372.35 |
392.61 |
408.21 |
429.21 |
|
±SD |
21.09 |
25.46 |
26.61 |
30.62 |
31.33 |
35.64 |
||
n |
12 |
12 |
12 |
12 |
12 |
12 |
||
G2, M & 100 |
Mean |
333.49 |
348.67 |
366.07 |
382.71 |
394.48 |
414.61 |
|
±SD |
20.91 |
24.45 |
30.62 |
31.50 |
33.73 |
39.05 |
||
n |
12 |
12 |
12 |
12 |
12 |
12 |
||
G3, M & 300 |
Mean |
333.86 |
349.97 |
361.35 |
384.41 |
399.83 |
423.79 |
|
±SD |
20.50 |
18.44 |
22.25 |
23.24 |
26.15 |
27.88 |
||
n |
12 |
12 |
12 |
12 |
12 |
12 |
||
G4, M & 1000 |
Mean |
331.99 |
357.47 |
378.38 |
396.73 |
411.93 |
426.47 |
|
±SD |
18.56 |
18.42 |
19.85 |
21.46 |
21.62 |
23.80 |
||
n |
12 |
12 |
12 |
12 |
12 |
12 |
M: Male; SD: Standard Deviation; n: Number of Animals
TABLE 2 (Contd..,). SUMMARY OF BODY WEIGHT (g) RECORD
Group, Sex & Dose (mg/kg body weight/day) |
Body Weight (g) on Days |
||||
1 |
7 |
14 |
21# |
||
G1, F & 0 |
Mean |
263.38 |
269.72 |
279.18 |
292.40 |
±SD |
16.71 |
16.23 |
19.53 |
22.58 |
|
n |
12 |
12 |
12 |
4 |
|
G2, F & 100 |
Mean |
263.21 |
269.81 |
281.42 |
292.27 |
±SD |
16.04 |
13.15 |
13.93 |
21.16 |
|
n |
12 |
12 |
12 |
5 |
|
G3, F & 300 |
Mean |
263.38 |
269.54 |
281.01 |
281.16 |
±SD |
15.85 |
13.80 |
16.37 |
21.01 |
|
n |
12 |
12 |
12 |
6 |
|
G4, F & 1000 |
Mean |
262.96 |
269.34 |
286.13 |
284.89 |
±SD |
14.76 |
14.63 |
15.25 |
19.99 |
|
n |
12 |
12 |
12 |
5 |
F: Female; SD: Standard Deviation; n: Number of Animals
#: The statistical analysis is not performed for day 21 body weight as the presented mean and standard deviations were calculated only for the females in cohabitation.
TABLE 3. SUMMARY OF PERCENTCHANGE IN BODY WEIGHT (%) WITH RESPECT TO DAY 1 RECORD
Group, Sex & Dose (mg/kg body weight/day) |
Percent Change in Body Weight (%) with respect to Day 1 during Days |
||||||
1 to 7 |
1 to 14 |
1 to 21 |
1 to 28 |
1 to 34 |
|||
G1, M & 0 |
Mean |
7.23 |
12.33 |
18.43 |
23.16 |
29.51 |
|
±SD |
2.62 |
4.24 |
5.26 |
5.83 |
7.60 |
||
n |
12 |
12 |
12 |
12 |
12 |
||
G2, M & 100 |
Mean |
4.55 |
9.74 |
14.78 |
18.31 |
24.26 |
|
±SD |
2.94 |
5.40 |
6.29 |
6.98 |
7.56 |
||
n |
12 |
12 |
12 |
12 |
12 |
||
G3, M & 300 |
Mean |
4.94 |
8.42 |
15.37 |
19.94 |
27.21 |
|
±SD |
3.94 |
6.65 |
7.45 |
7.38 |
9.22 |
||
n |
12 |
12 |
12 |
12 |
12 |
||
G4, M & 1000 |
Mean |
7.79 |
14.23 |
19.91 |
24.49 |
28.83 |
|
±SD |
4.61 |
7.57 |
9.96 |
10.01 |
9.93 |
||
n |
12 |
12 |
12 |
12 |
12 |
M: Male; SD: Standard Deviation; n: Number of Animals
TABLE 3 (Contd..,). SUMMARY OF PERCENTCHANGE IN BODY WEIGHT (%) WITH RESPECT TO DAY 1RECORD
Group, Sex & Dose (mg/kg body weight/day) |
Percent Change in Body Weight (%) with respect to Day 1 during Days |
|||
1 to 7 |
1 to 14 |
1 to 21# |
||
G1, F & 0 |
Mean |
2.45 |
6.02 |
9.31 |
±SD |
1.97 |
3.71 |
5.11 |
|
n |
12 |
12 |
4 |
|
G2, F & 100 |
Mean |
2.59 |
7.10 |
11.07 |
±SD |
2.35 |
5.26 |
4.99 |
|
n |
12 |
12 |
5 |
|
G3, F & 300 |
Mean |
2.42 |
6.86 |
8.90 |
±SD |
3.00 |
6.13 |
6.84 |
|
n |
12 |
12 |
6 |
|
G4, F & 1000 |
Mean |
2.45 |
8.87 |
11.94 |
±SD |
1.38 |
3.28 |
8.06 |
|
n |
12 |
12 |
5 |
F: Female; SD: Standard Deviation; n: Number of Animals
#: The statistical analysis is not performed for day 21 body weight as the presented mean and standard deviations were calculated only for the females in cohabitation.
TABLE 4. SUMMARY OFFEED CONSUMPTION (g/rat/day)RECORD
Group, Sex & Dose |
Feed Consumption (g) |
||||
Week 1 (Day 1 to 7) |
Week 2 (Day 7 to 14) |
Post Mating (Day 28 to 34) |
|||
G1, M & 0 |
Mean |
23.12 |
22.81 |
29.41 |
|
±SD |
1.34 |
2.28 |
0.67 |
||
n |
12 |
12 |
12 |
||
G2, M & 100 |
Mean |
22.76 |
23.08 |
28.95 |
|
±SD |
1.71 |
1.95 |
0.76 |
||
n |
12 |
12 |
12 |
||
G3, M & 300 |
Mean |
22.66 |
23.23 |
29.49 |
|
±SD |
1.48 |
2.35 |
0.66 |
||
n |
12 |
12 |
12 |
||
G4, M & 1000 |
Mean |
24.16 |
24.26 |
29.68 |
|
±SD |
1.12 |
1.73 |
0.66 |
||
n |
12 |
12 |
12 |
M: Male;SD: Standard Deviation; n: Number ofanimals
TABLE 4 (Contd..,). SUMMARY OF FEED CONSUMPTION (g/rat/day)RECORD
Group, Sex & Dose |
Feed Consumption (g) |
|||
Week 1 (Day 1 to 7) |
Week 2 (Day 7 to 14) |
|||
G1, F & 0 |
Mean |
18.19 |
18.43 |
|
±SD |
1.13 |
1.55 |
||
n |
12 |
12 |
||
G2, F & 100 |
Mean |
18.85 |
18.65 |
|
±SD |
1.93 |
1.58 |
||
n |
12 |
12 |
||
G3, F & 300 |
Mean |
17.63 |
18.21 |
|
±SD |
1.98 |
0.86 |
||
n |
12 |
12 |
||
G4, F & 1000 |
Mean |
18.33 |
18.60 |
|
±SD |
1.18 |
0.80 |
||
n |
12 |
12 |
F: Female; SD: Standard Deviation; n: Number of animals
TABLE 5. SUMMARY OF ABSOLUTE ORGAN WEIGHT (g) RECORD
Group, Sex & Dose |
Epididymes |
Testes |
Prostate+Seminal vesicles with coagulating glands (PSC) |
Thyroid along with parathyroid# |
|||||||
G1, M & 0 |
Mean |
1.7248 |
3.3488 |
3.8053 |
0.0357 |
||||||
±SD |
0.1932 |
0.2725 |
0.5705 |
0.0033 |
|||||||
n |
12 |
12 |
12 |
12 |
|||||||
G2, M & 100 |
Mean |
1.7994 |
3.3460 |
3.7837 |
0.0370 |
||||||
±SD |
0.1389 |
0.2603 |
0.3461 |
0.0022 |
|||||||
n |
12 |
12 |
12 |
12 |
|||||||
G3, M & 300 |
Mean |
1.6893 |
3.2506 |
4.1135 |
0.0359 |
||||||
±SD |
0.1685 |
0.2056 |
0.2480 |
0.0027 |
|||||||
n |
12 |
12 |
12 |
12 |
|||||||
G4, M & 1000 |
Mean |
1.7526 |
3.3840 |
4.1498 |
0.0363 |
||||||
±SD |
0.1925 |
0.2029 |
0.4138 |
0.0020 |
|||||||
n |
12 |
12 |
12 |
12 |
M: Male;SD: Standard Deviation; n: Number of animals: #: Weighed post fixation
TABLE 5 (Contd..,). SUMMARY OF ABSOLUTE ORGAN WEIGHT (g) RECORD
Group, Sex & Dose |
|
Uterus |
Thyroid along with parathyroid# |
||||
G1, F & 0 |
Mean |
0.6154 |
0.0291 |
||||
±SD |
0.0395 |
0.0031 |
|||||
n |
11 |
11 |
|||||
G2, F & 100 |
Mean |
0.5932 |
0.0253 |
||||
±SD |
0.0270 |
0.0033 |
|||||
n |
11 |
11 |
|||||
G3, F & 300 |
Mean |
0.6264 |
0.0279 |
||||
±SD |
0.0764 |
0.0056 |
|||||
n |
11 |
11 |
|||||
G4, F & 1000 |
Mean |
0.5864 |
0.0306 |
||||
±SD |
0.0420 |
0.0048 |
|||||
n |
12 |
12 |
F: Female;SD: Standard Deviation; n: Number of dams;
#: Weighed post fixation (excluding non-pregnant animals)
TABLE 6. SUMMARY
OF FASTING BODY WEIGHT (g) AND
ORGAN WEIGHT (%) RELATIVE TO FASTING BODY WEIGHT RECORD
Group, Sex & Dose (mg/kg body weight/day) |
Fasting Body Weight (g) |
Epididymes |
Testes |
Prostate+Seminal vesicles with coagulating glands (PSC) |
Thyroid along with parathyroid |
||||||||
G1, M & 0 |
Mean |
403.62 |
0.4300 |
0.8341 |
0.9467 |
0.0089 |
|||||||
±SD |
34.33 |
0.0578 |
0.0859 |
0.1466 |
0.0011 |
||||||||
n |
12 |
12 |
12 |
12 |
12 |
||||||||
G2, M & 100 |
Mean |
391.26 |
0.4644 |
0.8645 |
0.9795 |
0.0095 |
|||||||
±SD |
37.96 |
0.0638 |
0.1260 |
0.1606 |
0.0010 |
||||||||
n |
12 |
12 |
12 |
12 |
12 |
||||||||
G3, M & 300 |
Mean |
397.31 |
0.4275 |
0.8208 |
1.0396 |
0.0091 |
|||||||
±SD |
26.57 |
0.0570 |
0.0674 |
0.0932 |
0.0008 |
||||||||
n |
12 |
12 |
12 |
12 |
12 |
||||||||
G4, M & 1000 |
Mean |
399.57 |
0.4393 |
0.8503 |
1.0407 |
0.0091 |
|||||||
±SD |
24.64 |
0.0473 |
0.0805 |
0.1104 |
0.0008 |
||||||||
n |
12 |
12 |
12 |
12 |
12 |
M: Male;SD: Standard Deviation; n: Number of animals
TABLE 6 (Contd..,). SUMMARY OF FASTING BODY WEIGHT (g) AND
ORGAN WEIGHT (%) RELATIVE TO FASTING BODY WEIGHT RECORD
Group, Sex & Dose (mg/kg body weight/day) |
|
Fasting Body Weight |
Uterus |
Thyroid along with parathyroid |
|||||
G1, F & 0 |
Mean |
304.24 |
0.2035 |
0.0096 |
|||||
±SD |
21.64 |
0.0224 |
0.0013 |
||||||
n |
11 |
11 |
11 |
||||||
G2, F & 100 |
Mean |
306.66 |
0.1943 |
0.0083 |
|||||
±SD |
23.92 |
0.0148 |
0.0012 |
||||||
n |
11 |
11 |
11 |
||||||
G3, F & 300 |
Mean |
324.21 |
0.1933 |
0.0087 |
|||||
±SD |
28.55 |
0.0177 |
0.0022 |
||||||
n |
11 |
11 |
11 |
||||||
G4, F & 1000 |
Mean |
306.69 |
0.1921 |
0.0101 |
|||||
±SD |
24.23 |
0.0182 |
0.0020 |
||||||
n |
12 |
12 |
12 |
F: Female;SD: Standard Deviation; n: Number of animals
TABLE 7. SUMMARY RECORD OF VAGINAL SMEAR EXAMINATION FOR DETERMINATION OF OESTRUS CYCLICITY
Group & Dose |
No. of Females |
No. of Females with Regular Oestrus Cyclicity during Pre-mating, mating and on Lactation day 14 |
No. of Females with Irregular Oestrus Cyclicity during Pre-mating, mating and on Lactation day 14 |
||||
G1 & 0 |
12 |
12 |
0 |
||||
G2 & 100 |
12 |
12 |
0 |
||||
G3 & 300 |
12 |
12 |
0 |
||||
G4 & 1000 |
12 |
12 |
0 |
TABLE 8. SUMMARY OF COHABITATION RECORD
Group & Dose |
Copulatory Interval (Days) |
|
Conceiving Days (1 to 5) |
Conceiving Days (6 to 20) |
|||||||
G1 & 0 |
Mean |
5.83 |
n |
8 |
4 |
||||||
±SD |
4.63 |
||||||||||
n |
12 |
% |
66.7 |
33.3 |
|||||||
G2 & 100 |
Mean |
7.00 |
n |
6 |
6 |
||||||
±SD |
4.24 |
||||||||||
n |
12 |
% |
50.0 |
50.0 |
|||||||
G3 & 300 |
Mean |
7.42 |
n |
5 |
7 |
||||||
±SD |
4.60 |
||||||||||
n |
12 |
% |
41.7 |
58.3 |
|||||||
G4 & 1000 |
Mean |
7.58 |
n |
6 |
6 |
||||||
±SD |
4.80 |
||||||||||
n |
12 |
% |
50.0 |
50.0 |
SD: Standard Deviation; n: Number of animals; %: Percent
TABLE 9. SUMMARY RECORD OF BODY WEIGHT (g) DURINGGESTATION PERIOD
Group & Dose |
Body weight (g) on Gestation Days |
||||
0 |
7 |
14 |
20 |
||
G1 & 0 |
Mean |
283.86 |
297.35 |
342.70 |
421.08 |
±SD |
21.63 |
21.53 |
28.88 |
20.97 |
|
n |
11 |
11 |
11 |
11 |
|
G2 & 100 |
Mean |
291.15 |
304.38 |
349.07 |
422.35 |
±SD |
15.19 |
13.65 |
19.69 |
17.65 |
|
n |
11 |
11 |
11 |
11 |
|
G3 & 300 |
Mean |
291.93 |
307.30 |
364.18* |
440.73 |
±SD |
11.96 |
11.79 |
12.56 |
22.28 |
|
n |
11 |
11 |
11 |
11 |
|
G4 & 1000 |
Mean |
292.90 |
305.82 |
358.55 |
429.48 |
±SD |
14.68 |
15.66 |
17.10 |
24.61 |
|
n |
12 |
12 |
12 |
12 |
SD: Standard Deviation; n: Number of dams
*: Statistically significant than the control group (p<0.05)
TABLE 10. SUMMARY RECORD OF PERCENT CHANGE IN BODY WEIGHT (%) DURING GESTATION PERIOD
Group & Dose |
Percent Change in body weight (%) during Gestation |
|||
0 to 7 |
7 to 14 |
14 to 20 |
||
G1 & 0 |
Mean |
4.78 |
15.24 |
23.31 |
±SD |
0.82 |
4.95 |
6.98 |
|
n |
11 |
11 |
11 |
|
G2 & 100 |
Mean |
4.59 |
14.70 |
21.17 |
±SD |
1.38 |
4.36 |
5.18 |
|
n |
11 |
11 |
11 |
|
G3 & 300 |
Mean |
5.28 |
18.57 |
21.02 |
±SD |
1.78 |
3.46 |
4.51 |
|
n |
11 |
11 |
11 |
|
G4 & 1000 |
Mean |
4.41 |
17.30 |
19.82 |
±SD |
0.92 |
2.71 |
4.74 |
|
n |
12 |
12 |
12 |
SD: Standard Deviation; n: Number of dams
TABLE 11. SUMMARY RECORD OF FEED CONSUMPTION (g/dam/day) DURING GESTATION PERIOD
Group, Sex & Dose |
Feed Consumption (g) during Gestation |
||||
GD 0 to 7 |
GD 7 to 14 |
GD 14 to 20 |
|||
G1 & 0 |
Mean |
22.64 |
26.85 |
30.28 |
|
±SD |
1.35 |
1.46 |
1.12 |
||
n |
11 |
11 |
11 |
||
G2 & 100 |
Mean |
23.92* |
27.62 |
30.43 |
|
±SD |
0.79 |
0.96 |
0.91 |
||
n |
11 |
11 |
11 |
||
G3 & 300 |
Mean |
23.19 |
27.05 |
30.00 |
|
±SD |
0.89 |
1.49 |
1.06 |
||
n |
11 |
11 |
11 |
||
G4 & 1000 |
Mean |
23.92* |
26.79 |
30.47 |
|
±SD |
0.78 |
1.12 |
0.69 |
||
n |
12 |
12 |
12 |
SD: Standard Deviation; n: Number of
dams
*: Statistically significant than the control group (p<0.05)
TABLE 12. SUMMARY RECORD OF BODY WEIGHT (g) DURING LACTATION PERIOD
Group, Sex & Dose |
Body Weight (g) on Lactation Day |
||||
1 |
4 |
7 |
13 |
||
G1 & 0 |
Mean |
309.57 |
313.98 |
320.23 |
327.06 |
±SD |
21.76 |
22.19 |
24.05 |
21.94 |
|
n |
11 |
11 |
11 |
11 |
|
G2 & 100 |
Mean |
307.28 |
313.82 |
318.42 |
329.52 |
±SD |
30.13 |
28.65 |
28.85 |
23.41 |
|
n |
11 |
11 |
11 |
11 |
|
G3 & 300 |
Mean |
327.46 |
335.18 |
341.73 |
346.58 |
±SD |
35.21 |
31.80 |
31.26 |
28.52 |
|
n |
11 |
11 |
11 |
11 |
|
G4 & 1000 |
Mean |
306.28 |
313.07 |
318.52 |
327.51 |
±SD |
33.02 |
28.48 |
27.29 |
24.06 |
|
n |
12 |
12 |
12 |
12 |
SD: Standard Deviation; n: Number of animals
TABLE 13. SUMMARY RECORD OF PERCENT CHANGE IN BODY WEIGHT (%) DURING LACTATION PERIOD
Group, Sex & Dose |
Percent Change in body weight (%) during Lactation |
|||
1 to 4 |
4 to 7 |
7 to 13 |
||
G1 & 0 |
Mean |
1.42 |
1.97 |
2.20 |
±SD |
0.46 |
1.22 |
1.28 |
|
n |
11 |
11 |
11 |
|
G2 & 100 |
Mean |
2.20 |
1.48 |
3.70 |
±SD |
1.85 |
0.49 |
3.39 |
|
n |
11 |
11 |
11 |
|
G3 & 300 |
Mean |
2.50 |
1.98 |
1.50 |
±SD |
2.26 |
0.89 |
1.15 |
|
n |
11 |
11 |
11 |
|
G4 & 1000 |
Mean |
2.42 |
1.79 |
2.93 |
±SD |
2.73 |
0.64 |
1.70 |
|
n |
12 |
12 |
12 |
SD: Standard Deviation; n: Number of dams
TABLE 14. SUMMARY RECORD OF FEED CONSUMTION (g/dam/day) DURING LACTATION PERIOD
Group & Dose |
Feed Consumption (g) during Lactation |
||||
LD 1 to 4 |
LD 4 to 7 |
LD 7 to 13 |
|||
G1 & 0 |
Mean |
23.34 |
27.53 |
32.34 |
|
±SD |
1.26 |
1.55 |
1.22 |
||
n |
11 |
11 |
11 |
||
G2 & 100 |
Mean |
24.65 |
29.18 |
33.01 |
|
±SD |
2.32 |
2.43 |
1.04 |
||
n |
11 |
11 |
11 |
||
G3 & 300 |
Mean |
22.80 |
28.77 |
33.44 |
|
±SD |
1.89 |
3.14 |
2.17 |
||
n |
11 |
11 |
11 |
||
G4 & 1000 |
Mean |
24.67 |
28.77 |
33.64 |
|
±SD |
3.09 |
2.52 |
1.33 |
||
n |
12 |
12 |
12 |
SD: Standard Deviation; n: Number of dams; LD: Lactation Day
TABLE 15. SUMMARY RECORD OF GESTATION LENGTH (DAYS) AND DELIVERY DATA
Group & Dose |
|
Gestation Length (Days) |
|
Delivery Data |
||||||||||
Litter Size (No.) |
Live Pups (No.) |
Dead Pups (No.) |
|
Sex Ratio (M/F) at Birth |
Live Birth Index |
|||||||||
Total (No.) |
Male (No.) |
Female (No.) |
Total (No.) |
Male (No.) |
Female (No.) |
|||||||||
G1 & 0 |
Mean |
22.64 |
12.64 |
12.55 |
5.91 |
6.64 |
0.09 |
0.00 |
0.09 |
1.13 |
99.30 |
|||
±SD |
1.03 |
2.06 |
2.07 |
1.76 |
2.84 |
0.30 |
0.00 |
0.30 |
0.71 |
2.32 |
||||
n |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
||||
G2 & 100 |
Mean |
22.55 |
13.73 |
13.45 |
7.00 |
6.45 |
0.27 |
0.09 |
0.18 |
1.14 |
97.85 |
|||
±SD |
1.13 |
2.15 |
2.30 |
1.95 |
1.51 |
0.47 |
0.30 |
0.40 |
0.42 |
3.74 |
||||
n |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
||||
G3 & 300 |
Mean |
22.73 |
13.00 |
12.82 |
6.36 |
6.45 |
0.18 |
0.00 |
0.18 |
1.16 |
98.52 |
|||
±SD |
1.42 |
1.95 |
2.04 |
1.86 |
2.21 |
0.40 |
0.00 |
0.40 |
0.69 |
3.31 |
||||
n |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
||||
G4 & 1000 |
Mean |
22.83 |
11.83 |
11.83 |
5.58 |
6.25 |
0.00 |
0.00 |
0.00 |
0.92 |
100.00 |
|||
±SD |
1.27 |
2.17 |
2.17 |
2.15 |
0.87 |
0.00 |
0.00 |
0.00 |
0.38 |
0.00 |
||||
n |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
M: Male; F: Female;SD: Standard Deviation; n: Number of dams; NAD: No abnormality detected; LD: Lactation Day
TABLE 16. SUMMARY OF LITTER OBSERVATION RECORD DURING LACTATION PERIOD
Group & Dose |
|
No. of Live Pups At Birth |
During LD 1 to 4 |
Sex Ratio (M/F) LD 4 |
No. of Survived Pups during LD 1 to 4 |
Pup Survival Index LD 1 to 4 |
||||||
Live Pups (No.) |
|
Dead Pups (No.) |
||||||||||
Total (No.) |
Male (No.) |
Female (No.) |
Total (No.) |
Male (No.) |
Female (No.) |
|||||||
G1 & 0 |
Mean |
12.55 |
12.36 |
5.91 |
6.45 |
0.18 |
0.00 |
0.18 |
1.15 |
11.33 |
98.48 |
|
±SD |
2.07 |
2.20 |
1.76 |
2.81 |
0.60 |
0.00 |
0.60 |
0.70 |
4.14 |
5.03 |
||
n |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
12 |
11 |
||
G2 & 100 |
Mean |
13.45 |
13.45 |
7.00 |
6.45 |
0.00 |
0.00 |
0.00 |
1.14 |
12.33 |
100.00 |
|
±SD |
2.30 |
2.30 |
1.95 |
1.51 |
0.00 |
0.00 |
0.00 |
0.42 |
4.46 |
0.00 |
||
n |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
12 |
11 |
||
G3 & 300 |
Mean |
12.82 |
12.82 |
6.36 |
6.45 |
0.00 |
0.00 |
0.00 |
1.16 |
11.75 |
100.00 |
|
±SD |
2.04 |
2.04 |
1.86 |
2.21 |
0.00 |
0.00 |
0.00 |
0.69 |
4.18 |
0.00 |
||
n |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
12 |
11 |
||
G4 & 1000 |
Mean |
11.83 |
11.83 |
5.58 |
6.25 |
0.00 |
0.00 |
0.00 |
0.92 |
11.83 |
100.00 |
|
±SD |
2.17 |
2.17 |
2.15 |
0.87 |
0.00 |
0.00 |
0.00 |
0.38 |
2.17 |
0.00 |
||
n |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
M: Male: F: Female;SD: Standard Deviation; n: Number of dams; LD: Lactation Day
TABLE 16 (Contd..,). SUMMARY OF LITTER OBSERVATION RECORD DURING LACTATION PERIOD
Group & Dose day) |
Live Pups (No.) on LD 4 |
|
Pups Sacrificed for Blood Collection on LD 4 (No.) |
|
Live Pups (No.) on LD 4 after Sacrificed for Blood Collection |
During LD 4 to 7 |
|
Sex Ratio (M/F) at |
No. of Survived Pups during LD 4-7 |
Pup Survival Index |
|||||||||||||
Live Pups (No.) |
|||||||||||||||||||||||
Male (No.) |
Female (No.) |
Total (No.) |
Male (No.) |
Female (No.) |
Total (No.) |
Male (No.) |
Female (No.) |
Total (No.) |
Male (No.) |
Female (No.) |
Total (No.) |
|
Male (No.) |
Female (No.) |
Total (No.) |
||||||||
G1 & |
Mean |
5.91 |
6.45 |
12.36 |
0.00 |
1.36 |
1.36 |
5.91 |
5.09 |
11.00 |
5.91 |
5.09 |
11.00 |
0.00 |
0.00 |
0.00 |
1.62 |
11.00 |
100.00 |
||||
±SD |
1.76 |
2.81 |
2.20 |
0.00 |
0.81 |
0.81 |
1.76 |
2.47 |
1.55 |
1.76 |
2.47 |
1.55 |
0.00 |
0.00 |
0.00 |
1.32 |
1.55 |
0.00 |
|||||
n |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
|||||
G2 & |
Mean |
7.00 |
6.45 |
13.45 |
0.00 |
1.64 |
1.64 |
7.00 |
4.82 |
11.82 |
7.00 |
4.82 |
11.82 |
0.00 |
0.00 |
0.00 |
1.65 |
11.82 |
100.00 |
||||
±SD |
1.95 |
1.51 |
2.30 |
0.00 |
0.67 |
0.67 |
1.95 |
1.47 |
1.83 |
1.95 |
1.47 |
1.83 |
0.00 |
0.00 |
0.00 |
0.81 |
1.83 |
0.00 |
|||||
n |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
|||||
G3 & |
Mean |
6.36 |
6.45 |
12.82 |
0.00 |
1.55 |
1.55 |
6.36 |
4.91 |
11.27 |
6.36 |
4.91 |
11.27 |
0.00 |
0.00 |
0.00 |
1.75 |
11.27 |
100.00 |
||||
±SD |
1.86 |
2.21 |
2.04 |
0.00 |
0.82 |
0.82 |
1.86 |
2.12 |
1.49 |
1.86 |
2.12 |
1.49 |
0.00 |
0.00 |
0.00 |
1.29 |
1.49 |
0.00 |
|||||
n |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
|||||
G4 & 1000 |
Mean |
5.58 |
6.25 |
11.83 |
0.00 |
1.00 |
1.00 |
5.58 |
5.25 |
10.83 |
5.58 |
5.25 |
10.83 |
0.00 |
0.00 |
0.00 |
1.17 |
10.83 |
100.00 |
||||
±SD |
2.15 |
0.87 |
2.17 |
0.00 |
0.95 |
0.95 |
2.15 |
1.06 |
1.40 |
2.15 |
1.06 |
1.40 |
0.00 |
0.00 |
0.00 |
0.64 |
1.40 |
0.00 |
|||||
n |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
M: Male: F: Female;SD: Standard Deviation; n: Number of dams; LD: Lactation Day
TABLE 16 (Contd..,). SUMMARY OF LITTER OBSERVATION RECORD DURING LACTATION PERIOD
Group & Dose |
Live Pups (No.) on LD 7 |
During LD 7 to 13 |
|
Sex Ratio (M/F) at LD 7 |
No. of Survived Pups during LD |
Pup Survival Index |
||||||||||
Live Pups (No.) |
||||||||||||||||
Male (No.) |
Female (No.) |
Total (No.) |
Male (No.) |
Female (No.) |
Total (No.) |
Male (No.) |
Female (No.) |
Total (No.) |
||||||||
G1 & 0 |
Mean |
5.91 |
5.09 |
11.00 |
5.91 |
5.09 |
11.00 |
0.00 |
0.00 |
0.00 |
1.62 |
11.00 |
100.00 |
|||
±SD |
1.76 |
2.47 |
1.55 |
1.76 |
2.47 |
1.55 |
0.00 |
0.00 |
0.00 |
1.32 |
1.55 |
0.00 |
||||
n |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
||||
G2 & 100 |
Mean |
7.00 |
4.82 |
11.82 |
7.00 |
4.82 |
11.82 |
0.00 |
0.00 |
0.00 |
1.65 |
11.82 |
100.00 |
|||
±SD |
1.95 |
1.47 |
1.83 |
1.95 |
1.47 |
1.83 |
0.00 |
0.00 |
0.00 |
0.81 |
1.83 |
0.00 |
||||
n |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
||||
G3 & 300 |
Mean |
6.36 |
4.91 |
11.27 |
6.36 |
4.91 |
11.27 |
0.00 |
0.00 |
0.00 |
1.75 |
11.27 |
100.00 |
|||
±SD |
1.86 |
2.12 |
1.49 |
1.86 |
2.12 |
1.49 |
0.00 |
0.00 |
0.00 |
1.29 |
1.49 |
0.00 |
||||
n |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
||||
G4 & 1000 |
Mean |
5.58 |
5.25 |
10.83 |
5.58 |
5.25 |
10.83 |
0.00 |
0.00 |
0.00 |
1.17 |
10.83 |
100.00 |
|||
±SD |
2.15 |
1.06 |
1.40 |
2.15 |
1.06 |
1.40 |
0.00 |
0.00 |
0.00 |
0.64 |
1.40 |
0.00 |
||||
n |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
M: Male: F: Female;SD: Standard Deviation; n: Number of dams; LD: Lactation Day
TABLE 17. SUMMARY OF PUP OBSERVATIONS RECORD DURING LACTATION PERIOD
Group & Dose (mg/kg body weight/day) |
Pup Observation on LD 1 |
Pup Observation on LD 4 |
Pup Observation on LD 7 |
Pup Observation on LD 13 |
|
G1 & 0 |
N |
N |
N |
N |
|
G2 & 100 |
N |
N |
N |
N |
|
G3 & 300 |
N |
N |
N |
N |
|
G4 & 1000 |
N |
N |
N |
N |
|
LD: Lactation Day; N: Normal
TABLE 18. SUMMARY RECORD OF MEAN PUP WEIGHT (g) DURING LACTATION PERIOD
Group& Dose |
Mean Pup weight (g) on LD 1 |
|
Mean Pup weight (g) on LD 4 |
Mean Pup Weight (g) on |
Mean Pup weight (g) on LD 13 |
|||||||
Male |
Female |
Male |
Female |
Male |
Female |
Male |
Female |
|||||
G1 & 0 |
Mean |
6.68 |
5.98 |
9.99 |
8.52 |
14.55 |
12.15 |
25.65 |
21.90 |
|||
±SD |
0.49 |
0.66 |
0.76 |
0.74 |
1.12 |
1.26 |
1.63 |
2.36 |
||||
n |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
||||
G2 & 100 |
Mean |
6.24 |
5.30 |
10.26 |
8.89 |
14.99 |
13.13 |
27.22 |
22.26 |
|||
±SD |
0.68 |
0.83 |
1.15 |
1.11 |
1.54 |
1.82 |
2.26 |
2.16 |
||||
n |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
||||
G3 & 300 |
Mean |
6.39 |
5.76 |
10.22 |
8.65 |
15.63 |
13.13 |
27.11 |
21.93 |
|||
±SD |
0.59 |
0.66 |
1.41 |
0.83 |
1.14 |
1.62 |
1.48 |
3.26 |
||||
n |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
||||
G4 & 1000 |
Mean |
6.52 |
5.60 |
11.19* |
9.06 |
16.03* |
13.22 |
27.29 |
21.68 |
|||
±SD |
0.64 |
0.44 |
1.22 |
1.36 |
1.19 |
1.61 |
1.90 |
3.61 |
||||
n |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
SD: Standard Deviation; n: Number of dams;LD: Lactation Day
*: Statistically significant than the control group (p<0.05)
TABLE 19. SUMMARY OF UTERI OBSERVATIONS RECORD
Group & Dose |
No. of Corporaluetea |
No. of Implantations |
Pre-Implantation Loss |
Post-Implantation Loss |
Pre-natal Loss |
Post-natal Loss |
Post-natal Loss |
No. of Early Resorptions |
No. of |
|
G1 & 0 |
Mean |
12.82 |
12.64 |
1.22 |
0.70 |
0.09 |
1.52 |
0.18 |
0.00 |
0.00 |
±SD |
2.23 |
2.06 |
2.72 |
2.32 |
0.30 |
5.03 |
0.60 |
0.00 |
0.00 |
|
n |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
|
G2 & 100 |
Mean |
13.82 |
13.73 |
0.57 |
2.15 |
0.27 |
0.00 |
0.00 |
0.00 |
0.00 |
±SD |
2.23 |
2.15 |
1.88 |
3.74 |
0.47 |
0.00 |
0.00 |
0.00 |
0.00 |
|
n |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
|
G3 & 300 |
Mean |
13.18 |
13.00 |
1.36 |
1.48 |
0.18 |
0.00 |
0.00 |
0.00 |
0.00 |
±SD |
1.99 |
1.95 |
3.11 |
3.31 |
0.40 |
0.00 |
0.00 |
0.00 |
0.00 |
|
n |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
11 |
|
G4 & 1000 |
Mean |
12.00 |
11.83 |
1.28 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
±SD |
2.26 |
2.17 |
3.05 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
|
n |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
SD: Standard Deviation; n: Number of dams
TABLE 20. SUMMARY RECORD OF ANO-GENITAL DISTANCE [AGD] RATIO ON LACTATION DAY 4
Group & Dose |
|
Mean Male AGD Ratio |
Mean Female AGD Ratio |
|
G1 & 0 |
Mean |
2.53 |
1.17 |
|
±SD |
0.18 |
0.07 |
||
n |
11 |
11 |
||
G2 & 100 |
Mean |
2.66 |
1.17 |
|
±SD |
0.16 |
0.09 |
||
n |
11 |
11 |
||
G3 & 300 |
Mean |
2.65 |
1.18 |
|
±SD |
0.11 |
0.07 |
||
n |
11 |
11 |
||
G4 & 1000 |
Mean |
2.69 |
1.23 |
|
±SD |
0.21 |
0.17 |
||
n |
12 |
12 |
SD: Standard Deviation; n: Number of dams
TABLE 21. SUMMARY RECORD OF MALE PUP NIPPLE/AREOLAE RETENTION (no.) ON LACTATION DAY 13
Group & Dose |
No. of Nipples/Areolae |
||
G1 & 0 |
Mean |
0.00 |
|
±SD |
0.00 |
||
n |
12 |
||
G2 & 100 |
Mean |
0.00 |
|
±SD |
0.00 |
||
n |
11 |
||
G3 & 300 |
Mean |
0.00 |
|
±SD |
0.00 |
||
n |
11 |
||
G4 & 1000 |
Mean |
0.00 |
|
±SD |
0.00 |
||
n |
11 |
SD: Standard Deviation; n: Number of dams
TABLE 22. SUMMARY OF SERUM T4 LEVELS (ng/mL) RECORD - MALES
Group, Sex & Dose (mg/kg body weight/day) |
Serum T4 Levels (ng/mL) |
||
G1, M & 0 |
Mean |
101.492 |
|
±SD |
18.172 |
||
n |
12 |
||
G2, M & 100 |
Mean |
84.759* |
|
±SD |
5.744 |
||
n |
12 |
||
G3, M & 300 |
Mean |
82.176* |
|
±SD |
8.380 |
||
n |
12 |
||
G4, M & 1000 |
Mean |
73.659* |
|
±SD |
5.815 |
||
n |
12 |
M: Male; SD: Standard Deviation; n: Number of animals
*: Statistically significant than the control group (p<0.05)
TABLE 23. SUMMARY OF SERUM T4 LEVELS (ng/mL) RECORD - FEMALES
Group, Sex & Dose (mg/kg body weight/day) |
Serum T4 Levels (ng/mL) |
||
G1, F & 0 |
Mean |
78.821 |
|
±SD |
11.933 |
||
n |
11 |
||
G2, F & 100 |
Mean |
73.067 |
|
±SD |
7.682 |
||
n |
11 |
||
G3, F & 300 |
Mean |
64.148* |
|
±SD |
5.037 |
||
n |
11 |
||
G4, F & 1000 |
Mean |
67.385* |
|
±SD |
8.707 |
||
n |
12 |
SD: Standard Deviation; n: Number of dams
*: Statistically significant than the control group (p<0.05)
TABLE 24. SUMMARY OF SERUM T4 LEVELS (ng/mL) RECORD - LACTATION DAY 4 PUPS
Group & Dose |
Serum T4 Levels (ng/mL) |
||
G1 & 0 |
Mean |
70.985 |
|
±SD |
5.708 |
||
n |
9 |
||
G2 & 100 |
Mean |
69.599 |
|
±SD |
3.939 |
||
n |
10 |
||
G3 & 300 |
Mean |
69.114 |
|
±SD |
3.762 |
||
n |
9 |
||
G4 & 1000 |
Mean |
70.672 |
|
±SD |
5.016 |
||
n |
7 |
SD: Standard Deviation; n: Number of dams (Serum collected from pups with litter size of more than ten per dam pooled for analysis)
TABLE 25. SUMMARY OF SERUM T4 LEVELS (ng/mL) RECORD - LACTATION DAY 13 PUPS
Group & Dose |
Serum T4 Levels (ng/mL) |
||
G1 & 0 |
Mean |
73.460 |
|
±SD |
8.044 |
||
n |
11 |
||
G2 & 100 |
Mean |
71.633 |
|
±SD |
7.751 |
||
n |
11 |
||
G3 & 300 |
Mean |
71.633 |
|
±SD |
7.751 |
||
n |
11 |
||
G4 & 1000 |
Mean |
75.088 |
|
±SD |
7.169 |
||
n |
12 |
SD: Standard Deviation; n: Number of dams (Serum collected from pups per dam pooled for analysis
TABLE 26. SUMMARY OF GROSS PATHOLOGY RECORD
Organs/Lesions |
Group |
G1 |
G2 |
G3 |
G4 |
||||
Dose (mg/kg body weight/day) |
0 |
100 |
300 |
1000 |
|||||
Sex |
M |
F |
M |
F |
M |
F |
M |
F |
|
No. of Animals |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
|
No Abnormality Detected |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
Note: The numeral indicates number of animal/s without lesion. M: Male; F: Female
TABLE 27. SUMMARY OF HISTOPATHOLOGY RECORD
Group No. |
G1 |
G4 |
||
Dose (mg/kg body weight/day) |
0 |
1000 |
||
Sex |
M |
F |
M |
F |
No. of Animals |
12 |
12 |
12 |
12 |
ORGANS AND OBSERVATIONS |
||||
EPIDIDYMIDES |
|
|
|
|
Interstitial MNC infiltration ; unilateral |
1 |
X |
1 |
X |
Note: The numeral indicates number of animal/s with lesion; M: Male; F: Female; X: Organs not subjected for histopathological examination due to sex difference.
Effect on fertility: via oral route
- Endpoint conclusion:
- no adverse effect observed
- Dose descriptor:
- NOAEL
- 1 000 mg/kg bw/day
- Study duration:
- subacute
- Species:
- rat
- Quality of whole database:
- Study is an OECD 421 guideline conform GLP study.
Effect on fertility: via inhalation route
- Endpoint conclusion:
- no study available
Effect on fertility: via dermal route
- Endpoint conclusion:
- no study available
Effects on developmental toxicity
Effect on developmental toxicity: via inhalation route
- Endpoint conclusion:
- no study available
Effect on developmental toxicity: via dermal route
- Endpoint conclusion:
- no study available
Justification for classification or non-classification
There is no evidence to suggest that a classification for reproductive toxicity is appropriate.
With reference to the OECD 421 studies performed with the registration substance, it is concluded that the test item is not subject to classification and labelling according to Regulation 1272/2008/EC regarding reproductive toxicity.
Additional information
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