7-amino-3-{(E)-[5-({4-(2-chloroethyl)butanoyl}amino)]diazenyl}-4-hydroxy-8-[(E)-(4-{2-(sulfonatooxy)ethyl}) diazenyl]naphthalene, polysulfonate, polysulfonyl, polyphenyl, sodium/potassium salt

Toxicity to reproduction

Administrative data

Endpoint:

screening for reproductive / developmental toxicity

Remarks:

based on test type (migrated information)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16 May 2012 to 01 July 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study performed under GLP compliance

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

(Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany)

Limit test:

no

Test material

Test material information

Constituent 1

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST SYSTEM:
Young healthy male and nulliparous, non pregnant female rats [strain: Wistar rat Crl:WI(Han)] (Full-Barrier), were used in this study. The animals were derived from a controlled full barrier maintained breeding system (SPF) (Source: Charles River, 97633 Sulzfeld, Germany)
According to Art. 9.2, No.7 of the German Act on Animal Welfare the animals were bred for experimental purposes.
At the start of treatment the age of the animals was 10-11 weeks. The range of the body weight was:
Females: 184.6-210.6 g, (mean: 199.79g, ± 20%= 39.96 g)
Males: 262.2-302.6 g, (mean: 281.89 g, ± 20%= 56.38 g)

HOUSING AND FEEDING CONDITIONS:
- Full barrier in an air-conditioned room
- Temperature: 22 3 °C
- Relative humidity: 55 10%
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: 10 x / hour
- Free access to Altromin 1324 maintenance diet for rats and mice (Lot. No. 2355)
- Free access to tap water, sulphur acidified to a pH of approximately 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals)
- The animals were housed individually in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding (Lot. No. 261111) except during mating period when individual male and female rats were cohabited.
- Certificates of food, water and bedding are filed at BSL BIOSERVICE.
- Adequate acclimatisation period (at least five days)

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

The test item was administered daily during 14 days pre mating and 14 days mating period in both male and in female rats, during gestation period and up to post natal day 3 in female rats. The male rats were dosed until the minimum total dosing period of 28 days is completed.
The test item was administered by gavage using a gavaging canula. The maximum dose volume administered was 5 mL / kg body weight.
For each animal the individual dosing volume was calculated on the basis of the most recently measured body weight.

Details on mating procedure:

Animals were mated in the ratio of 1:1 (Male to Female). The subsequent morning and the next morning then onwards the vaginal smear of females were checked to confirm the pregnancy. The day of sperm positive vaginal smear was considered as gestation day (GD) 0.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Each dosing concentration was analysed for nominal concentration. Homogeneity of the test item in the vehicle was analyzed for the low and high dose concentrations.
Samples for the nominal concentration verification were taken in study week 1 (first week of pre-mating period), week 3 (first week of mating), week 5 (gestation) and week 7 (gestation/lactation).
Samples for homogeneity were taken from the top, middle and bottom of the HD and LD preparation in study week 1 and 5.
All concentration samples were stored frozen (approximately -20°C) until the analysis was performed.
The dose formulation analysis was performed at BSL Bioservice Scientific Laboratories GmbH. The results are reported in the annexure of the final report.

Duration of treatment / exposure:

Males: 28 days; Females: approx. 54 days

Frequency of treatment:

7 days/ week

Details on study schedule:

Arrival of the Test Item: 28 November 2012
Date of Draft Study Plan: 03 May 2012
Date of Final Study Plan: 08 May2012
Date of Start of Experiment: 16 May 2012
Date of End of Experiment: 15 May 2011
Date of Start of Delegated Phase (Histopathology): 04 August 2012
Date of End of Delegated Phase (Histopathology): 21 September 2012
Date of Start of Delegated
Phase (Formulation Analysis) 05 September 2012
Date of End of Delegated
Phase (Formulation Analysis) 21 September 2012
Date of Date of Draft Phase Report (Histopathology): 25 September 2012
Date of Final Phase Report (Histopathology): 05 November 2012
Date of Draft Phase Report (Formulation Analysis): 19 October 2012
Date of Final Phase Report (Formulation Analysis): 28 November 2012
Date of Study Completion: 09 December 2012

Doses / concentrations

Remarks:

Doses / Concentrations:
100, 300, 1000
Basis:
nominal conc.

No. of animals per sex per dose:

10 animals/sex/dose

Control animals:

yes, concurrent vehicle

Details on study design:

Number and sex of the animals
80 animals (40 males and 40 females) were included in the study (10 male and 10 female animals per group). The study included three dose groups (LD, MD and HD) and one control group (C).

Preparation of the animals
The rats were assigned to the dose/control groups using a randomization procedure based on stratified body weight to ensure harmonized group mean body weights between the groups. Each animal was assigned a unique identification number and caged individually. The animals were acclimatised for at least five days before the first dose administration.

Dosage
In consultation with the sponsor the doses 100, 300, 1000 were selected for the 3 dose groups (LD, MD and HD):
The highest dose level was chosen with the aim of inducing toxic effects, but not death or severe suffering. Thereafter, a descending sequence of dose levels was selected with a view to demonstrate any dose-related response and a NOAEL.
The animals in the control group were handled in an identical manner to the test group subjects and received aqua ad injectionem (sterile water) the same volume as used for the treatment groups.


Administration of doses
The test item was administered daily during 14 days pre mating and 14 days mating period in both male and in female rats, during gestation period and up to post natal day 3 in female rats. The male rats were dosed until the minimum total dosing period of 28 days is completed.
The test item was administered by gavage using a gavaging canula. The maximum dose volume administered was 5 mL / kg body weight.
For each animal the individual dosing volume was calculated on the basis of the most recently measured body weight.
On treatment day 8 most of the HD group animals (males 31-40 and females 71-75) received the dose concentration higher than 1000 mg/kg body weight. Considering the overall result of the study this deviation did not impact the validity of the study.

Mating
Animals were mated in the ratio of 1:1 (Male to Female). The subsequent morning and the next morning then onwards the vaginal smear of females were checked to confirm the pregnancy. The day of sperm positive vaginal smear was considered as gestation day (GD) 0. Cages were arranged in such a way that possible effects due to cage placement are minimised.
Females showing no evidence of copulation up to 14 day mating period were sacrificed 26 days after the last day of the mating period.

Clinical observation
General clinical observations were made twice a day except during weekend and holidays where observation was made daily once, approximately at the same time each day and considering the peak period of anticipated effects after dosing.

Body weight and food Consumption
All animals were weighed at randomisation, male rats weighed weekly during the entire study period and at termination. Females were weighed weekly during pre mating period, on GD 0, 7, 14, 20 and on PND 0 (within 24 hours of parturition) and PND 4 along with pups.
The food consumption was measured on corresponding day of body weight after the beginning of the dose administration. The food consumption was not measured during mating period in both male and female rats.

Litter observations
The duration of gestation was recorded and was calculated from day 0 of pregnancy. Each litter was examined as soon as possible after delivery of the dam to establish the number and sex of pups, stillbirths, live births, runts and the presence of gross abnormalities.
Live pups were counted and sexed. Litters were weighed within 24 hours of parturition (day 0 post-partum) and on day 4 post-partum. Live pups were identified by marking with tattoo. In addition to the observations on parent animals, any abnormal behavior of the offspring was recorded.

Pathology
Gross necropsy
Males were sacrificed after the completion of mating period (total dosing of 28 days), pregnant females were sacrificed on respective post natal day 4 along with pups and non pregnant females sacrificed on their respective day 26 after the evidence of mating or completion of mating period by using Ketamine/Xylazine (2:1) at the dose volume of 1.4 ml/kg body weight. At the time of sacrifice the adult animals were examined macroscopically for any abnormalities or pathological changes. Special attention was paid to the organs of the reproductive system.
Dead pups and pups killed at day 4 post-partum were carefully examined for gross external abnormalities.
The number of implantation sites and corpora lutea was recorded in all pregnant females by gross observations.
The ovaries, uterus with oviduct and cervix, vagina, testes, epididymides, accessory sex organs (prostate, seminal vesicles with coagulating glands as a whole), and all organs showing macroscopic lesions of all adult animals were preserved in 10 % neutral buffered formalin. Testes and epididymides were fixed in modified Davidson’s Solution for 24 hours and then transferred to 10 % neutral buffered formalin.

Organ weight
The testes and epididymides of all male adult animals and ovaries, uterus with oviduct and cervix of all female adult animals were weighed. Paired organs were weighed separately.

Histopathology
Full histopathology was carried out on the preserved organs/ tissues of all animals in the control and high dose groups. All organs and tissues listed in table 2 were evaluated in C and HD group animals. In addition, testis and epididymis was also evaluated in MD group animals. Macroscopic lesions were evaluated in all study animals.
For paired organs marked with (*), both sides were examined.

Tissues evaluated microscopically
Cervix
Seminal vesicle *
Coagulating gland *
Ovary *
Testis *
Uterus
Epididymis *
Vagina
Prostate gland
Macroscopic lesions
For testes, a detailed qualitative examination was made; taking into account the tubular stages of the spermatogenic cycle at evaluation of additional haematoxylin-PAS (Periodic Acid Schiff) stained slides.
All gross lesions were examined microscopically.
Processing and histopathological evaluation was performed at GLP-certified test site Propath UK Ltd Willow Court, Netherwood Road, GB - Hereford HR2 6JU and KALEIDIS –Consultancy in Histopathology, 6 rue du Gers, 68300 Saint-Louis, France. Blocking, embedding, cutting, staining and professional evaluation was performed according to the corresponding SOPs of the test site.

Examinations

Parental animals: Observations and examinations:

Body weight, food consumption, clinical signs, pathology, organ weight (reproductive organs), histopathology (reproductive organs)

Oestrous cyclicity (parental animals):

Not examined

Sperm parameters (parental animals):

Not Examined

Litter observations:

The duration of gestation was recorded and was calculated from day 0 of pregnancy. Each litter was examined as soon as possible after delivery of the dam to establish the number and sex of pups, stillbirths, live births, runts and the presence of gross abnormalities.
Live pups were counted and sexed. Litters were weighed within 24 hours of parturition (day 0 post-partum) and on day 4 post-partum. Live pups were identified by marking with tattoo. In addition to the observations on parent animals, any abnormal behavior of the offspring was recorded.

Postmortem examinations (parental animals):

yes

Postmortem examinations (offspring):

not examined

Statistics:

For statistical analysis one-way analysis of variance (ANOVA) followed by Dunnett’s multiple comparison test was carried out to reveal any differences between control- and test groups. Statistical analysis was performed with GraphPad Prism (version V) software and p<0.05 was considered as statistical significants.

Reproductive indices:

Copulation, fertility, delivery indices

Offspring viability indices:

yes

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Other effects:

not examined

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

Details on results (P0)

Day of sacrifice
All male animals were sacrificed on day 29. Non pregnant females were sacrificed on the respective day 26 after the sperm positive vaginal smear as an evidence of mating. Lactating females along with pups were sacrificed on respective post natal day 4.

Clinical observation and mortality
There were very few clinical signs recorded in control and treated group animals during the study period, which are as follows,
Males: C, LD and MD groups- no findings; HD group- moderate salivation (1/10 animals), slight piloerection (4/10 animals), moving the bedding (1/10 animals), red urine (1/5 animals), dark coloured nasal discharge (1/10 animals), dark coloured feces (1/10 animals).
Females: C- Alopecia on back (1/10 animals); LD group- small wound on snout (1/10 animals); MD group- Alopecia on shoulder, forelimbs and abdomen (3/10 animals) and Eschar on shoulder and snout (1/10 animals); HD group- no findings).
In addition to above findings all males and females from HD group had red colored skin and urine. These clinical findings observed in male and female animals were not considered to be of toxicological relevance.
There were no mortalities observed in male and female animals during the study period.

Body weight and body weight change
In males and females, statistical analysis of body weight data revealed no effect on body weight throughout the study period in treated groups when compared with controls. However, there was decrease in body weight change noted in male HD group between the mating/ post mating days 14-21. This decrease was considered to be incidental in origin.

Food consumption
In males and females, statistical analysis of food consumption data revealed no effect on food consumption throughout the study period in treated groups when compared to control.

Precoital interval and duration of gestation
There was no treatment related effect observed on the duration of gestation and precoital interval in the treated groups when compared with controls. However, the precoital interval was slightly higher in treated groups, but without reaching the statistical significance.
All females in control and treated groups showed evidence of copulation during 14 days mating period. Successful mating of females resulted in pregnancy rate as follows, Control and LD groups: 90%; MD and HD groups: 100%.

Gross pathology
At necropsy, male (after minimum total dosing of 28 days) and female rats (on post-natal day 4) using Ketamine/Xylazine (2:1) at the dose volume of 1.4 mL/kg body weight.
In males, various macroscopic findings observed were reddish discoloured mesenteric lymph nodes (C:4/10, LD:4/10, MD:8/10, HD 7/10 animals), two dark points (holes) on left kidneys (C:2/10, LD:6/10, MD:10/10, HD 10/10 animals), yellow spot ø 0,3cm on right epididymidis (C: 1/10), yellow spot ø 0,5cm on left epididymidis (C: 1/10), discoloured axillary lymph nodes (LD:1/10, MD:3/10, HD 7/10 animals), reddish discoloured kidneys (LD:5/10, MD:10/10, HD 10/10 animals), yellow spot ø 0,7cm on right epididymidis (MD:1/10 animals), discoloured skin/ mammary gland (MD:6/10, HD 9/10 animals), reddish discoloured testes (MD:2/10, HD 4/10 animals), reddish discoloured lung: (MD:1/10 animals).
In females, reddish discoloured mesenteric lymph nodes (LD: 1/10, MD: 3/10 animals), reddish discoloured kidneys (LD: 1/10, MD: 2/10, HD: 7/10 animal), reddish discoloured skin (MD: 1/10 animal), enlarged spleen (MD: 1/10 animal), reddish discoloured thymus (HD: 1/10 animal), discoloured adrenal glands (HD: 2/10 animal), reddish discoloured uterus with oviduct and cervix (HD: 3/10 animal), reddish discoloured axillary Lymph nodes (HD: 1/10 animal), reddish discoloured ovaries (HD: 3/10 animal), reddish discoloured vagina (HD: 3/10 animal), reddish discoloured implantation sites (HD: 3/10 animal).
The reddish discoloration of organs in treatment groups were considered to be attributable to the colour of the test item. Other macroscopic organ findings were very few and not considered to be test item-related.

Organ weight
In males, statistically significant increase in absolute and relative (to body weight) of Prostate (including Seminal vesicle with coagulating gland) weight was observed in HD group. Microscopic examination of Prostate (including Seminal vesicle with coagulating gland) revealed no changes. Hence, this difference in organ weight was considered less likely to be adverse.
There were no treatment related effects observed for testes and epididymides weights in treated groups compared to the controls.
In females, there were no statistically significant differences observed in the absolute organ weight and relative organ weight to terminal body weight of the treated groups when compared with the controls.

Histopathology
No histopathological findings considered to be of toxicological relevance were seen in this study.
In the male and female reproductive organs, findings related to test item administration were restricted to the presence of minor amounts of brown or red-brown pigment, mainly seen in macrophages, in MD and HD groups. The pigment was considered to be due to test item colour. Under the conditions of this study, it was considered to be without toxicological significance, in view of the low degree of severity observed and the absence of any changes indicating functional impairment of the organs concerned. Therefore, these organs were not further evaluated in the low dose group (testis, epididymis) or in the low and medium dose group (other male and female reproductive organs).
In the kidney, red-brown pigment deposition in the corticotubular epithelium was observed in a dose-related manner in MD and HD groups and was considered due to test item colour. Furthermore, some minor degenerative renal changes were seen in a low number of males or females in HD group (tubular degeneration, debrisfilled tubules), and minor degrees of basophilic (regenerative tubules) occurred in a proportion of animals in MD and HD groups. However, as these latter changes may occasionally be seen spontaneously in rats of this strain and age, and as kidney was not systematically evaluated in control rats of this study, a test item relationship could not be established under the conditions of this study. Furthermore, no indication of renal histopathology (other than pigment deposition) had been observed in a 28-day repeated dose oral toxicity study in Wistar rats performed with the same dosages and at the same test facility (BSL Study No. 115998).
Minimal brown pigment deposition was also seen in a number of lymph nodes from animals in MD and HD groups, but was not accompanied by any functional impairment of these organs.

Dose formulation analysis
Concentration analysis of formulation samples was determined in study week 1, 3, 5
and 7 for all dose groups. The mean recoveries observed in LD, MD and HD groups
were 95.7%, 99.3% and 100.4% of the nominal concentration, respectively.
Stability of formulation samples was investigated in study week 1 for LD and HD dose groups. After 6 hours storage at -20°C recovery compared to starting value was between 96.1% and 121.2%.
Homogeneity of formulation samples was determined in study week 1 and 5 for LD and HD dose groups. The mean recovery observed for LD dose group was between 90.3 and 96.6% of the nominal value, and between 100.1 and 100.3% for HD dose group. The coefficients of variation of the different sampling locations (top, middle, bottom) were between 1.8 and 6.9% in LD dose group, and between 1.5 and 6.1% in HD dose group.

Effect levels (P0)

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: see 'Remark'

Remarks on result:

other: Generation: P and F1 Generation (migrated information)

Results: F1 generation

General toxicity (F1)

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

not examined

Histopathological findings:

not examined

Details on results (F1)

Litter weight data
The statistical analysis of litter weight data of treated and control groups measured on PND 0 and PND 4 revealed no treatment related effect on group mean litter weight, total litter weight, male litter weight and female litter weight.

Pre and post natal data
The group means of corpora lutea, implantation sites, live pups born on PND 0, percent pre implantation loss and percent post implantation loss in the treated groups remained unaffected due to treatment when compared with corresponding controls.

Litter data
There was no treatment related effect observed on the number of male pups, number of female pups, sex ratio, live pups, still birth and runt on PND 0 and total number of live pups and sex ratio on PND 4.

Reproductive indices
The copulation index, fertility index and viability index remained unaffected due to treatment in treated groups compared to corresponding control.
All pregnancies resulted in normal births and therefore delivery index remained unaffected in all treated groups. 1/10 control female and 1/10 female treated at 100 mg/kg/day were found not to be pregnant at terminal sacrifice, but this was not considered to be test item-related.

Pup survival data
The survival of the pups from PND 0 to PND 4 remained unaffected due to the treatment in all treated groups compared to control. However, 1 pup from animal 59 (LD group) was found dead on PND 1, which was considered to be incidental.

Pup external findings
There were no treatment related gross external findings observed in pups from any of the treated groups on PND 0 and 4.

Overall reproductive toxicity

Reproductive effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

In conclusion, the repeated dose administration of FAT 40858/A TE in sterile water to the male (28-29 days) and female (maximum 54 days) Wistar rats at dosages of 100, 300 and 1000 mg/kg body weight revealed no major toxicological findings.

Based on the data generated from this reproduction/ developmental toxicity screening test with FAT 40858/A TE, the no observed adverse effect level (NOAEL) is considered to be 1000 mg/kg body weight for reproduction/ developmental toxicity screening in males and females.

Executive summary:

The aim of this study was to assess the possible effect of FAT 40858/A TE on male, female fertility and embryofetal development in Wistar rats.

In this study, four groups comprised of 10 adult male and 10 non pregnant nulliparous female rats [Wistar Crl:WI(Han)] were dosed daily by oral gavage with 100, 300 and 1000 mg/kg body weight per day of FAT 40858/A TE at dose volume of 5 mL/kg body weight. The test item was formulated in sterile water with an administration volume of 5 mL/kg body weight. Control animals were handled identically as treated groups and received sterile water in similar volume as treated groups.

Animals were examined daily for the clinical signs and mortality. Body weight and food consumption was measured weekly except during the mating period.

After 14 days of treatment to both male and female, animals were paired (1:1) for maximum 14 days. The subsequent morning onwards, vaginal smears of females were checked to confirm the evidence of mating. After the confirmation of the mating, females were separated. Each litter was examined as soon as possible after delivery of the dam to establish the number and sex of pups, stillbirths, live births, runts and the presence of gross abnormalities. Live pups were counted, sexed and litters weighed within 24 hours of parturition and on day 4 post-partum.

Males and females were sacrificed on day 29 and post natal day 4 respectively and subjected to necropsy. Non pregnant females were sacrificed on their respective day 26 after the evidence of mating.

Clinical Observation and Mortality:

There were no predominant clinical signs considered to be due to toxicity observed in treatment groups compared to the controls.

There were no mortalities observed in males or females during the study period.

Body Weight Development :

In males and females, statistical analysis of body weight data revealed no effect on body weight throughout the study period in treated groups when compared with controls.

Food Consumption:

In males and females, statistical analysis of food consumption data revealed no effect on food consumption throughout the study period in treated groups when compared with controls.

Litter Weight data:

The statistical analysis of litter weight data of treated and control groups measured on PND 0 and PND 4 revealed no treatment related effect on group mean litter weight, total litter weight, male litter weight and female litter weight.

The test item formulation was prepared freshly and administered daily during 14 days pre mating and 14 days mating period in both males and in females, during gestation period and up to post natal day 3 in females. Males were dosed for 28 days. Dose volumes were adjusted weekly based on the most recent body weight measurement.

Precoital interval and duration of gestation:

There was no treatment related effect observed for the duration of gestation and precoital interval the in treated groups when compared with controls. All females in control and treated groups showed evidence of copulation during 14 days mating period. Successful mating of females resulted pregnancy rate as follows, Control and LD groups: 90%; MD and HD groups: 100%.

Pre and post natal data:

Group mean of corpora lutea, implantation sites, live pups born on PND 0, percent pre implantation loss and post implantation loss in treated groups remained unaffected due to treatment when compared with controls.

Litter data:

There was no treatment related effect observed on total number of pups born, number of male pups, number of female pups, sex ratio, live pups, still birth and runt on PND 0 and total number of live pups and sex ratio on PND 4.

Reproductive indices:

The copulation index, fertility index and viability index in treated groups remained unaffected due to treatment when compared to the control. All pregnancies resulted in normal births and therefore delivery index remained unaffected in all treated groups. 1/10 control female and 1/10 female treated at 100 mg/kg/day were found not to be pregnant at terminal sacrifice, but this was not considered to be test item-related.

Pup survival data:

Survival of the pups from PND 0 to PND 4 remained unaffected due to the treatment in all treated groups compared to the control.

Pup External findings:

There were no treatment related gross external findings observed in pups from the treated groups on PND 0 and 4.

Gross Pathology:

At terminal sacrifice, reddish discoloration of various organs were noted in a dose-related manner in all treated groups. In LD group, reddish discoloration was noted for kidney in 6/10 males and 1/10 female. In MD group, reddish discoloration was seen in the kidney of 10/10 males and 2/10 females. Skin, testis and some other organs were also reddish discolored in some animals of MD group. In HD group, reddish discoloration was observed in all males and females, most often in numerous organs. Reddish discolorations were considered to be due to the test item color. Other macroscopic organ findings were very few and not considered to be test item-related, including a yellow spot in the epididymis of 2/10 control males and 1/10 male of LD group, considered to represent spontaneous spermatic granuloma(s).

Organ Weight:

In males and females, there were no treatment related changes observed in the absolute and relative organ weights of the treated groups when compared with the controls.

Histopathology :

No histopathological findings considered to be of toxicological relevance were seen in this study, based on the evaluation of a limited number of organs only.