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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
one-generation reproductive toxicity
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study performed to GLP according to OECD Guidelines for the testing of chemicals No. 415

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 415 [One-Generation Reproduction Toxicity Study (before 9 October 2017)]
Deviations:
no
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
other: liquid
Details on test material:
Name : Cyclamen Aldehyde
CAS # : 103-95-7
CAS name : Benzenepropanal, α-methyl-4-(1-methylethyl)-
EC # : 203-161-1
EC name : 3-p-cumenyl-2methylpropionaldehyde
IUPAC name : 2-methyl-3-[4-(propan-2-yl)phenyl]propanal
Batch No. : VE00051122

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
Twenty-five Crl:CD(SD) strain rats/sex/dose were administered the test material at dosages of 0, 25, 75 or 150 mg/kg/day in corn oil. Dose volume was 4 ml/kg which was adjusted daily on the basis of the individual bodyweights.
These treated male and treated female rats were cohabitated (for a maximum of 21 days) with untreated cohort male or female rats. Treated P generation male rats were 38 days old upon arrival and weighed 131-166 grams at study start. The untreated P generation male rats were 122 days old upon arrival and weighed 437-550 grams at study start. Treated P generation female rats were 66 days old upon arrival and weighed 218-285 grams at study start. Untreated P generation female rats were 66 days old upon arrival and weighed 224-311 grams at study start.

Conditions
The study rooms were maintained under conditions of positive airflow relative to a hallway and independently supplied with a minimum of 10 changes per hour of 100% fresh air that had been passed through 99.97% HEPA filters. Room temperature was maintained at 64 F to 79 F (18 C to 26 C); relative humidity was targeted at 30% to 70 %. P generation rats were housed individually in stainless steel, wire-bottomed cages, except during the cohabitation and postpartum periods. During cohabitation, each pair of male and female rats was housed in the male rat's cage. Beginning no later than DG 20, P generation female rats were individually housed in nesting boxes until they either naturally delivered litters or were sacrificed on DG 25. Each dam and delivered litter were housed in a common nesting box during the postpartum period. After weaning, F1 generation pups selected for continued evaluation were individually housed in stainless steel, wire-bottomed cages. A 12 hour light/dark cycle was maintained. Cage pan liners were changed at least 3 times per week. Cages were changed approximately every other week. Bedding was changed as often as necessary to keep the rats dry and clean. Rats were given ad libitum access to Certified Rodent Diet #5002 meal in individual feeders. Local water that had been processed by passage through a reverse osmosis membrane (R.O. water) was available to the rats ad libitum from an automatic watering access system and/or individual water bottles attached to the cages. Chlorine was added to the processed water as a bacteriostat. Bed-o'-cobs bedding was used as the nesting material. Chewable Nylabones were supplied to all rats during the study.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
Male P generation rats were gavaged once daily beginning 83 days prior to cohabitation, through cohabitation, continuing through the day before sacrifice. Female P generation rats were gavaged once daily beginning 14 days before cohabitation, through cohabitation and DG 25 (rats that did not deliver) or day 22 postpartum (rats that delivered a litter). F1 generation rats were not directly dosed but may have been exposed to the test material in utero during gestation and through maternal milk postpartum.
Details on mating procedure:
Treated or untreated female rats wtih spermatozoa observed in a smear of the vaginal contents and/or a copulatory plug observed in situ were considered to be at DG 0 and assigned to individual housing. Treated female rats not mated within the first 14 days of cohabitation were assigned alternate male rats that had mated (same dosage group) and remained in cohabitation for a maximum of 7 additional days. Treated male rats that did not mate an untreated female rat within the first 14 days of cohabitation were assigned an alternate untreated female rat and remained in cohabitation for a maximum of 7 additional days. Treated or untreated female rats not mated after completing of the 21 day cohabitation period were considered at DG 0 on the last day of cohabitation and assigned to individual housing. Three untreated female rats not mated with a treated male rat within the first 14 days of cohabitation were considered DG 0. Day 1 of lactation (postpartum) was defined as the day of birth and was also the first day of which all pups in a litter were individually weighed. All F1 generation rats were weaned on PPD 22, based on observed growth and viability of the pups. At weaning, 25 male and 25 female pups per group from treated dams (mated with untreated male rats) and 25 male and 25 female pups per group from untreated dams (mated withtreated male rats) were selected resulting in a total of 350 F1 generation rats (175 per sex) chosen for continued evaluation.
Analytical verification of doses or concentrations:
not specified
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
0 mg/kg bw /day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
25 mg/kg bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
75 mg/kg bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
150 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose:
Twenty-five Crl:CD(SD) strain rats/sex/dose were administered the test material at dosages of 0, 25, 75 or 150 mg/kg/day in corn oil. Dose volume was 4 ml/kg which was adjusted daily on the basis of the individual bodyweights.
These treated male and treated female rats were cohabitated (for a maximum of 21 days) with untreated cohort male or female rats. Treated P generation male rats were 38 days old upon arrival and weighed 131-166 grams at study start. The untreated P generation male rats were 122 days old upon arrival and weighed 437-550 grams at study start. Treated P generation female rats were 66 days old upon arrival and weighed 218-285 grams at study start. Untreated P generation female rats were 66 days old upon arrival and weighed 224-311 grams at study start.
Control animals:
yes, concurrent vehicle
Positive control:
No data

Examinations

Parental animals: Observations and examinations:
The following parameters were evaluated in the P generation rats (treated or untreated): viability, clinical observations, body weights, feed consumption, estrous cycling, mating and fertility, natural delivery and litter observations, sperm assessments (motility and concentration), organ weights, histopathology and/or necropsy observations.
Oestrous cyclicity (parental animals):
Estrous cycling was evaluated daily by examination of vaginal cytology beginning 28 days (treated female rats) or 14 days (untreated female rats) before scheduled cohabitation and continuing through cohabitation until mating was confirmed or until the end of the cohabitation period was reached.
Sperm parameters (parental animals):
To assess the potential toxicity of the test material in P generation male rats on the male reproductive system, organ weights, sperm evaluation and histopathology were evaluated. Sperm concentration and motility were evaluated.
Litter observations:
Treated rats were observed for clinical signs, abortions, premature deliveries and deaths daily before dosage administration, between one and two hours following dosage administration and at the end of the normal working day. Untreated rats were observed for clinical signs, abortions, premature deliveries and deaths weekly during the precohabitation and cohabitation periods, on DG 0, twice weekly during the gestation period and on days 1, 5, 8, 11, 15, 18 and 22 postpartum. In addition, treated and untreated female rats were evaluated for adverse clinical signs observed during parturition, duration of gestation (DG 0 to the day the first pup was observed), litter size (all pups delivered) and pup viability at birth. Treated and untreated dams and their respective litters were observed for maternal behaviour on days 1, 5, 8, 14, 18 and 22 postpartum.
. F1 generation pups were individually weighed and litters were examined after delivery to identify the number and sex of pups, stillbirths, live births and gross alterations. Anogenital distance was measured for all live F1 generation pups on days 1 and 22 postpartum. Nipple eruption was evaluated for all live F1 generation pups once on day 12 postpartum. F1 generation litters were observed at least twice daily for viability. The pups in each litter were counted once daily. Clinical observations were recorded once daily during he preweaning period. Pup body weights were recorded on days 1 (birth), 5, 8, 11, 15, 18 and 22 postpartum, and at least weekly during the postweaning period and on the day of sacrifice. During the postweaning period, rats were observed twice daily for viability and once daily for clinical observations and/or general appearance. Body weights and feed consumption values were recorded weekly during the postweaning period. Female F1 generation rats were evaluated for the age of vaginal patency, beginning on day 28 postpartum and continuing until this developmental parameter is achieved, or until day 40 postpartum.
Postmortem examinations (parental animals):
The body weight was recorded for each rat on the day the criterion was achieved/recorded. A terminal body weight was also recorded. Treated P generation male rats that died or were sacrificed before scheduled termination were examined for the cause of death or condition on the day the observation was made. The rats were examined for gross lesions and a complete necropsy was performed. Tissues were retained and/or microscopically evaluated, but not weighed. Surviving treated and untreated P generation male rats were sacrificed by CO2 asphyxiation after completion of the cohabitation period. Treated and untreated female rats that delivered a litter and their respective pups not selected for continued evaluation were sacrificed on day 22 of presumed lactation (DL 22). Female rats that did not deliver a litter were sacrificed on day 25 of gestation (DG 25). Male and female F1 generation rats weaned and selected for continued evaluation were sacrificed by CO2 asphyxiation on day 57, 58 or 60 postpartum. All surviving treated P generation male and female rats were subject to a complete necropsy examination which included:
evaluation of the carcass and musculoskeletal system;
all external surfaces and orifices;
cranial cavity and external surfaces of the brain and spinal column;
the nasal cavity and neck with associated organs and tissues;
and thoracic, abdominal, and pelvic cavities with their associated organs and tissues.
To assess the potential toxicity of the test material in P generation male rats on the male reproductive system, organ weights, sperm evaluation and histopathology were evaluated. Sperm concentration and motility were evaluated. Treated dams with no surviving pus were sacrificed after the last pup was found dead or missing and presumed cannibalized. The number and distribution of implantation sites were recorded. Uteri of apparently nonpregnant rats were examined while being pressed between glass plates to confirm the absence of implantation sites, and retained in 10% Neutral buffered formalin for microscopic evaluation. Tissues identified for microsocpic evalution were embedded in paraffin, sectioned, mounted on glass slides and stained with hematoxylin and eosin. Histopathological evaluation was performed by a board-certified veterinary pathologist. Untreated P generation male rats were sacrificed via CO2 asphyxiation after completion of the cohabitation period and carcasses were discarded without evaluation. Untreated P generation female rats were sacrificed via CO2 asphyxiation after completion of the 22 day postpartum period. The number and distribution of implantation sites were recorded. Untreated female rats that did not deliver a litter were sacrificed on DG 25 and examined for pregnancy status. Uteri of apparently non-pregnant rats were examined to confirm the absence of implantation sites. Female rats without a confirmed mating date that did not deliver a litter were sacrificed on an estimated DG 25. The carcasses of the untreated female rats were discarded without further evaluation.
Postmortem examinations (offspring):
Unscheduled deaths of F1 generation pups were evaluated as follows: pups that died before examination of the litter for pup viability were evaluated for vital status at birth. The lungs were removed and immersed in water. Pups with lungs that sunk were identified as stillborn; pups with lungs that floated were identified as liveborn and to have died shortly after birth. Pups with gross lesions were preserved in Bouin's solution for possible future evlaution. Pups that died or were sacrified before schedule termination were examined for gross lesions and the cuase of death or condition on the day the observation was made. Pups found on PPDs 2 to 5 were preserved in Bouin's solution for possible future evaluation. Pups found on days 6 to 22 postpartum were preserved in neutral buffered 10% formalin for possible future evaluation. Pups that died or were sacrified before scheduled termination from days 5 through 22 postpartum were examined for gross lesions and the cause of death or condition on the day the observation is made. All pups selected for continued evaluation were sacrified by CO2 asphyxiation on PPD 22 and examined for gross lesions; gross lesions were preserved in 10% NBF for possible future histopathological evaluation. Male and female F1 generation rats were sacrificed by CO2 asphyxiation on days 57, 58 and 60 postpartum. A gross necropsy of the thoracic, abdominal and pelvic viscera was performed. Gross lesions were preserved in 10% NBF. Tissues selected were weighed and preserved in 10% NBF. Statistical analyses were conducted on data.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not specified
Mortality:
not specified
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not specified
Other effects:
no effects observed

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
effects observed, treatment-related
Reproductive performance:
no effects observed

Details on results (P0)

In treated male rats, reproductive organ weights were reduced at both 75 and 150mg/kg bw/day. Adverse effects on sperm analyses and histopathological changes to the epididymides were observed at 150mg/kg bw/day dose level. In treated female rats, reduced gestational body weights were observed at 150mg/kg bw/day dose level. Reduced pup body weights were also observed at both 150mg/kg bw/day dose levels. In untreated females mated to the treated males, a reduced number of implantation sites and a reduced fertility index were observed at th 150mg/kg bw/day dose level.

Effect levels (P0)

open allclose all
Dose descriptor:
NOAEL
Effect level:
75 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
mortality
Remarks on result:
other: n/a
Dose descriptor:
NOAEL
Effect level:
25 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
female
Basis for effect level:
mortality
Remarks on result:
other: n/a

Results: F1 generation

General toxicity (F1)

Body weight and weight changes:
effects observed, treatment-related

Effect levels (F1)

open allclose all
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
75 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
mortality
Remarks on result:
other: n/a
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
25 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
female
Basis for effect level:
mortality
Remarks on result:
other: n/a

Overall reproductive toxicity

Reproductive effects observed:
not specified

Any other information on results incl. tables

150mg/kg body weight changes , Developmental effects , eye effects , liver , organ weight changes , reproductive effects

dose was mg/kg/day. P generation male treated rats results: no treatment-related deaths were observed. One male rat at this dose level was sacrificed on DS 53 as a result of a broken palate. In addition, three male rats in the vehicle control group were found dead on DS 86, 123, 130. None of these deaths were attributed to treatment with the test material because the incidence was not dosage-dependent. All other P generation male rats survived to scheduled sacrifice. Clinical signs attributed to the test material were limited to slight or moderate excess salivation at this dose level. This observation occurred intermittently during the dosage period and was not considered an adverse effect of the test material. Reductions bodyweight gain occurred intermittently during the dosage period prior to cohabitation, with statistically significant reductions occurring on DSs 50 to 57 and DSs 64 to 71. Body weight gains were also reduced at this dose level on DSs 84 to 92 at each tabulated interval between DS 99 and DS 134, in comparison to the vehicle control group values. Overall, body weight gains at this dose level were significantly reduced on DSs 1 to 134 and DS 1 to termination. The average body weight was slightly reduced beginning on DS 72 and continuing until scheduled sacrifice, reaching statistical significance on DS 120 when compared to the vehicle control group value. There were no apparent effects of the test material on feed consumption in the P generation male rats at this dose level. The test material caused infertility following mating with untreated female rats at this dose level. There was only one pregnancy (1/24) produced by matings at this dose level. Terminal body weights for P generation male rats at this dose level were slightly reduced, in comparison to the vehicle control group value. This reduction did not reach statistical significance, but reflected the statistically significant reduction in body weight gains that occurred in this dosage group for the cumulative dosage period. The test material increased the absolute and relative weights of the epididymides (left, right and cauda) at this dose level. The increased dpidiymal weights generally reflected the presence of masses on the cauda epidiymis and microscopic observations of moderate to marked sperm granulomas. In addition, the absolute and relative weights of the liver were increased or significantly increased at this dose level, as compared to the vehicle control group values. The absolute weight of the adrenal glands was significantly reduced at this dose level in comparison to the vehicle control group value. The decreased adrenal weights correlated microscopically with minimal adrenal cortical atrophy, affecting the zona fasciculate and zona reticualris. Sperm motility from sperm taken from the vas deferens could not be observed in all rats at this dose level. The motility values in each of the samples generally reflected the presence of drifting debris, headless sperm, detached heads and/or less than the required number of sperm for evaluation. The effects correlated with the infertility that was observed in the treated male rats that were mated with the untreated cohort female rats. Pregnancy occurred in 23 and 1 of the 25 to 28 untreated female rats that were assigned to mate with treated male rats in the 0 (vehicle) and 150 mg/kg/day dose level, respectively. The pregnant dam at this dose level did not deliver a litter. P generation female rat results: all rats survived to scheduled sacrifice. No clinical signs related to the test material were observed in the treated P generation female rats during the premating, gestation and lactation period. During the first week of the dosage period (DSs 1 to 8), body weight gains were significantly reduced as compared to the vehicle controls. Despite the rebound during the second week of the dosage period, body weight gains remained significantly reduced for the entire premating dosage period (DSs 1 to 15) as compared to the vehicle control group value. In gestating rats at this dose level, body weight gains were reduced at each tabulated interval within the gestation dosage period. Although the reductions within the gestation period did not reach statistical significance, the cululative material body weight gains (DGs 0 to 21) were significantly reduced as compared to the vehicle control group value. The average maternal body weight on DG 18 and DG 21 was significantly reduced at this dose level compared to the vehicle control group value. At the beginning o the lactation period (DLs 1 to 5), body weight losses were observed in each test material treated group compared to gains in the vehicle control group during the same period. The losses in body weight were statistically significant at this dose level. Weight gains remained significantly reduced during the next tabulated interval (DLs 5 to 8) compared to the vehicle control group value. The average maternal body weight on DLs 5 to 11 was significantly reduced at this dose level in comparison with the vehicle control group values. During the premating period, absolute and relative feed consumption values at this dose level were slightly, but significantly reduced on DSs 1 to 8 in comparison to the vehicle control group values. These reductions correlated with the statistically significant reductions in body weight gain at this dose level during the same period. These reductions in feed consumption were transient and did not persist during the gestation period. However, during the lactation period, absolute and relative feed consumption values were reduced (often statistically significant) at this dose level at each tabulated interval within the lactation period and overall for DLs 1 to 15 in comparison to the vehicle control group values. There were no apparent effects of the test material on estrous cyclicity or mating and fertility parameters at this dose level. Pregnancy occurred in 24 or 25 of the 25 mated female rats in the 0 (vehicle) and 150 mg/kg/day dose level, respectively. All pregnant dams delivered litters. The test material significantly reduced the average number of implantation sites per delivered litter at this dose level, in comparison to the vehicle control group value. However, the average value for implantation sites per delivered litter was within the range observed historically at the testing facility. Other effects on natural delivery at this dose level included: a significant increase in the number of dams with all pups dying between days 1 and 5 postpartum; a significant reduction in the average number of pups delivered per litter and the average number of liveborn pups per litter; a significant increase in the number of stillborn pups that were delivered; a significant increase in pup mortality on days 1 to 5 postpartum with a corresponding significant reduction in the viability index; and a significant reduction in the averages for the number of surviving pups per litter and the live litter size at weighing at on days 1 through 22 postpartum. The average values for the total number of delivered pups, the number of liveborn pups and the number of stillborn pups were within the range observed historically at the testing facility. In addition, the average pup body weight per litter was significantly reduced at this dose level between days 1 and 22 postpartum as compared to the vehicle control group values. There were no test material-related necropsy observations. Terminal body weights were comparable among the dosage groups and did not significantly differ. The test material significantly increased the absolute and relative weights of the liver in dams at this dose level. The absolute and relative weights of the non-gravid uterus (with the cervix) was significantly decreased and the weights of the left and right ovary were significantly decreased at this dose level as compared to the vehicle control group values. There were no microscopic changes in the uterus or ovaries that could be correlated with the differences in these organ weights. At this dose level, an increase in the number of primordial follicles was present in comparison to the corresponding control group. The increased primordial follicles may be inversely correlated to the decreased ovarian weights at this dose level because the smaller ovaries may cause a more concentrated dispersion of follicles than in the larger ovaries of the rats in the control group. However, since no microscopic correlated were reported for the histopathology of these rat ovaries, biological relevance of the increased number of follicles could not be completely defined. Corpora lutea were present in all rats evaluated. F1 generation pus of treated male rats mated with untreated female rats results: there were no litters produced at this dose level from mating of treated P generation male rats with untreated female rats. F1 generation pups of treated female rats mated with untreated male rats: a significant number of litters (20/24 litters) had one or more pups with a lenticular opacity in one or both eyes. The lenticular opacities were first observed on day 16 and generally persisted until day 22 postpartum and was confirmed during necropsy examination of pups not selected for continued evaluation. During the postweaning period, the number of F1 generation male and female rats with a lenticular opacity in one or both eyes was significantly increased at this dose level in comparison to the vehicle control group value. Ths observation was more prevalent in F1 generation male rats than in the female rats. IN male pups, anogenital distance on day 1 postpartum was not affected by treatment of P generation female rats with the test material. On day 22 postpartum, there was a significant reduction in the anogenital distance of male pups at this dose level in comparison to the vehicle control group.

 

25 mg/kg liver , no observed adverse effect level , organ weight changes dose was mg/kg/day. P generation treated male rats results at 25 mg/kg/day:

there were no treatment-related deaths. One male rat at this dose level was sacrificed on day 65 as a result of a broken palate. In addition, three male rats in the vehicle control group and one male at this dose level were found dead on DS 86, 105, 123, or 130. None of these deaths were attributed to treatment with the test material because the incidence was not dosage-dependent. All other P generation male rats survived to scheduled sacrifice. No clinical signs attributed to the test material were observed at this dose level. There were no apparent effects of the test material on body weight, feed consumption, organ weights, sperm motility, sperm count and density from the cauda epididymis at this dose level in P generation treated male rats. Pregnancy occurred in 23 and 24 of 25 to 28 untreated female rats that were assigned to mate with treated male rats in the 0 (vehicle) and 25 mg/kg/day dosage groups respectively. All pregnant dams in the 0 (vehicle) and 25 mg/kg/day dosage groups delivered litters. Natural delivery and litter observations were unaffected by dosages of the test material at this dose level. P generation female rats results at 25 mg/kg/day: all P generation treated female rats survived to scheduled sacrifice. No clinical signs related to the test material were observed during the premating, gestation and lactation periods. At the beginning of the lactation period (DLs 1 to 5) bodyweight losses were observed in the test material treated group compared to gains in the vehicle control group during the same period. Body weight gains rebounded in the test material treated group during the next tabulated interval (DLs 5 to 8). No effects were observed on food consumption, estrous cyclicity, mating or fertility parameters. Pregnancy occurred in 24 or 25 of the 25 mated female rats in the 0 (vehicle) and 25 mg/kg/day dosage groups, respectively (mated with untreated male rats). All pregnant dams delivered litters. Natural delivery and litter observations were unaffected by dosages of the test material at this dose level. There were no test material related necropsy observations. Terminal body weights were comparable among the dosage groups and did not signficantly differ. The test material significaintly increased the absolute and relative weights of the liver in this effect. Results of F1 generation pups of treated male rats mated with untreated female rats: none of the clinical signs that occurred in F1 generation pups during the preweaning or postweaning periods were attributed to treatment of P generation male rats with test material at this dose level. Anogenital distance, nipple eruption and sexual maturation were not affected by paternal treatment with the test material and there were no gross lesions in the F1 generation pups or rats that were attributed to the test material. Body weights (including terminal body weights), body weight gains and feed consumption values in the F1 generation male and female rats were unaffected by paternal treatment with the test material at this dose level. There were no test material related changes in the absolute or relative (% terminal weight) weight of the reproductive organs or the pituitary, brain or adrenal glands of the F1 generation male or female rats. Results from F1 generation pups of treated female rats mated with untreated male rats: in male pups, anogenital distance on day 1 postpartum was not affected by treatment of P generation female rats with test material at any dose level tested. In female pups, anogenital distance on day 1 postpartum was not affected by treatment of P generation female rats with the test material at this dose level. There were no gross lesions in the F1 generation pups or rats that were attributed to the test material at this dose level. There were no effects on F1 generation male or female rat body weights, body weight gains or feed consumption at this dose level when compared to vehicle control values. There were no effects on sexual maturation (preputial separation or vaginal opening) at any maternal dose level tested. Terminal bodyweights of F1 generation male and female rats were comparable at this dose level when compared to the vehicle control group. There were no test material substance-related changes in the absolute or relative (% terminal body weight) weight of the reproductive organs or the pituitary, brain or adrenal glands in either sex at any dosage tested. The reproductive NOAEL in the P generation male and female rats is 25 mg/kg/day. The NOAEL for general toxicity in P generation female rats is 25 mg/kg/day. The NOAEL for viability and growth of the F1 generation offspring of treated P generation female rats is 25 mg/kg/day.

75 mg/kg body weight changes , liver , no observed adverse effect level , organ weight Changes

dose was mg/kg/day. P generation treated male rats results at 55 mg/kg/day: there were no treatment-related deaths. Three male rats in the vehicle control group were found dead on DS 86, 123, and 130. None of these deaths were attributed to treatment with the test material. All other P generation male rats survived to scheduled sacrifice. No clinical signs attributed to the test material were observed at this dose level. There were no apparent effects of the test material on body weight, feed consumption at this dose level in P generation treated male rats. The absolute and relative weights of the liver were increased or significantly increased at this dose level as compared to the vehicle control group values. The absolute weight of the adrenal glands was significantly reduced at this dose level when compared to the vehicle control group. The decreased adrenal weights correlated microscopically with minimal adrenal cortical atrophy, affecting the zona fasciculate and zona reticularis. Sperm motility from sperm taken from the as deferens could not be observed in 13/25 rats at this dose level. The motility values in each of the samples generally reflected the presence of drifting debris, headless sperm, detached heads and/or less than the required number of sperm for evaluation. The test material significantly reduced the sperm count and density from the cauda epididymis as comapred to the vehicle treated group values. The average values for sperm historically at the testing facility. Pregnancy occurred in 23 of 25 to 28 untreated female rats that were assigned to mate with treated male rats in the 0 (vehicle) and 75 mg/kg/day dosage groups. All pregnant dams in the 0 (vehicle) and 75 mg/kg/day dosage groups delivered litters. Natural delivery and litter observations were unaffected by dosages of the test material at this dose level. P generation female rats results at 25 mg/kg/day: all P generation treated female rats survived to scheduled sacrifice. No clinical signs related to the test material were observed during the premating, gestation and lactation periods. During the first week of the dosage period (DSs 1 to 8) body weight gains were reduced at this dose level as compared to vehicle controls. Despite the rebound during the second week of the dosage period, body weight gains at this dose level remained reduced for the entire premating dosage period (DSs 1 to 15) as compared to the vehicle control group value. At the beginning of the lactation period (DLs 1 to 5) bodyweight losses were observed in the test material treated group compared to gains in the vehicle control group during the same period. Body weight gains rebounded in the test material treated group during the next tabulated interval (DLs 5 to 8). During the lactation period, absolute and relative feed consumption values were reduced (often statistically significant) at this dose level at each tabulated interval within the lactation period and overall for DLs 1 to 15 in comparison to the vehicle control group values. There were no apparent effects of the test material on estrous cyclicity, mating or fertility parameters at this dose level. Pregnancy occurred in 24 or 25 of the 25 mated female rats in the 0 (vehicle) and 75 mg/kg/day dosage groups, respectively (mated with untreated male rats). All pregnant dams delivered litters. Natural delivery were unaffected by dosages of the test material at this dose level. The average pup body weight per litter was significantly reduced at this dsoe level at each tabulated interval between days 1 and 22 postpartum as compared with the vehicle control values. There were no test material related necropsy observations. Terminal body weights were comparable among the dosage groups and did not signficantly differ. The test material significaintly increased the absolute and relative weights of the liver in dams treated at this dose level. There were no microscopic changes to correlate this effect. Results of F1 generation pups of treated male rats mated with untreated female rats: none of the clinical signs that occurred in F1 generation pups during the preweaning or postweaning periods were attributed to treatment of P generation male rats with test material at this dose level. Anogenital distance, nipple eruption and sexual maturation were not affected by paternal treatment with the test material and there were no gross lesions in the F1 generation pups or rats that were attributed to the test material. Body weights (including terminal body weights), body weight gains and feed consumption values in the F1 generation male and female rats were unaffected by paternal treatment with the test material at this dose level. There were no test material related changes in the absolute or relative (% terminal weight) weight of the reproductive organs or the pituitary, brain or adrenal glands of the F1 generation male or female rats. Results from F1 generation pups of treated female rats mated with untreated male rats: in male pups, anogenital distance on day 1 postpartum was not affected by treatment of P generation female rats with the test material at any dosage level tested. On day 22 postpartum, there was a significant reduction in the anogenital distance of male pups at this dose level in comparison to the vehicle control group value. Whe covaried with fetal body weights per litter, the statistically sifniciant reduction in anogenital distance was no longer apparent. This developmental delay correlated with an overall reduction in pup weights on day 22 postpartum. In female pups, anogenital distance on day 1 postpartum was not initially affected by treatment of P generation female rats with the test material at this dose level. There were no gross lesions in the F1 generation pups or rats that were attributed to the test material. Reflecting significant reductions in the average pup body weight per litter that occurred prior to weaning, the average body weight were also significantly reduced in the F1 generation male and female rats on days 23, 30, 37, 44, 51 and/or 57 postpartum at this dose level. There were no effects on sexual maturation (preputial separation or vaginal opening) at any maternal dose level tested. In the F1 generation male and female rats, terminal bodyweights were comparable among the dosage groups and did not significantly differ. There were no test material related changes in the absolute or relative (% terminal body weight) weight of the reproductive organsor the pituitary, brain or adrenal glands in either sex at any dose level tested. The NOAEL for general toxicity in treated males mated to untreated female is 75 mg/kg/day. The NOAEL for F1 viability and growth in treated males mated to untreated females is 75 mg/kg/day. body weight changes , Developmental effects , eye effects , liver , organ weight changes , reproductive effects dose was 150 mg/kg/day (cond't). F1 generation pups of treated female rats mated with untreated male rats: When covaried with fetal body weights per litter, the statistically significant reduction in anogenital distance was no longer apparent. This developmental delay correlated with an overall reduction in pup body weights on day 22 postpartum. In female pups, anogenital distance on day 1 postpartum was not initially affected by treatment of P generation female rats with the test material. However, when covaried with fetal body weights per litter, there was a statistically significant increase in anogenital distance at this dose level in comparison to the control group value. This increase in anogenital distance was no longer apparent by day 22 postpartum. There were no gross lesions in the F1 generation pups or rats that were attributed to the test material. In F1 generation male rats, body weight gains were significantly reduced at this maternal dose group on days 23 to 30 postpartum, days 30 to 37 postpartum and overall for the entire postweaning period (days 23 to 57 postpartum) in comparison to the vehicle control group values. Transient, but statistically significantly reductions in body weight gains occurred in the F1 generation female rats on day 23 to 30 postpartum in comparison to the vehicle control group value. Reflecting significant reductions in the average pup body weight per litter that occurred prior to weaning, the average body weight were also significantly reduced in the F1 generation male and female rats on days 23, 30, 37, 44, 51 and/or 57 postpartum at this dose level. Corresponding to significant reductions in body weight gains, absolute feed consumption values were significantly reduced in the F1 generation male rats on days 23 to 30 postpartum and days 30 to 37 postpartum and in F1 generation female rats on days 23 to 30 postpartum at this dose level in comparison to vehicle control values. Relative to body weight, F1 generation male and female rats consumed significantly more feed overall for the entire postweaning period (days 23 to 57 postpartum) at this dose level. There were no effects on sexual maturation (preputial separation or vaginal opening) at any maternal dosage tested. Terminal body weights in the F1 generation male rats were significantly reduced at this dose level in comparison to vehicle control group value. In the F1 generation female rats, terminal bodyweights were comaprable among the dosage groups and did not significantly differ. There were no test material related changes in the absolute or relative (% terminal body weight) weight of the reproductive organs or the pituitary, brain or adrenal glands in either sex at any dosage level tested.

Applicant's summary and conclusion

Conclusions:
The No-observable-adverse-effect-level (NOAEL) for general toxicity of the test material in P generation male rats is 75 mg/kg/day. The reproductive NOAEL in the P generation male rats is 25 mg/kg/day. The NOAEL for general toxicity of the test material in the P generation female rats is 25 mg/kg/day. The reproductive NOAEL in the P generation female rats is 25 mg/kg/day. The NOAEL for viability and growth of the F1 generation offspring of treated P generation male rats is 75 mg/kg/day. The NOAEL for viability and growth of the F1 generation offspring of treated P generation female rats is 25 mg/kg/day.
Executive summary:

Protocol

The reproductive effects of the test material in corn oil were evaluated in a previous study. This study was the basis for the dosage selection in the associated main reproductive study. Test material dosages of 0 (vehicle), 25, 75 and 150 mg/kg/day were administered via gavage to P generation male and female rats at a volume of 4 ml/kg. Treated rats were mated with untreated cohorts of male and female rats. Male rats were given the vehicle or test material once daily beginning 14 days before cohabitation, through

cohabitation, and continuing through the day before sacrifice, while female rats were given the vehicle or test material once daily beginning 14 days before cohabitation, through cohabitation and continuing through day 24 of presumed gestation (DG 24) (rats that did not deliver a litter), day 4 postpartum (DL 4 (rats that delivered a litter) or DS 44 (rats wtih no confirmed date of mating).

Comments

Number of male and female rats used per dose level not provided. No further details provided.

Results

Details

vehilce was corn oil

Summary

Based on the results of the study, test material dose levels of 25, 75 and 150 mg/kg/day in corn oil were selected for the associated reproductive study. The anticipated NOAELs for the males is 25 mg/kg/day, for the females is 75 mg/kg/day and for the offspring is 25 mg/kg/day.

150mg/kg

body weight changes , organ weight changes , reproductive effects dose was mg/kg/day. In treated male rats, reproductive organ weights were reduced at this dose level. Adverse effects on sperm analyses and histopathological changes to the epididymides were observed at this dose level. In treated female rats, reduced gestational body weights were observed at this dose level. Reduced pup body weights were also observed at this dose level. In untreated females mated to the treated males, a reduced number of implantation sites and a reduced fertility index were observed at this dose level.

25 mg/kg

no effects dose was mg/kg/day. No effects were reported.

75 mg/kg

body weight changes , organ weight changes , reproductive effects dose was mg/kg/day. In treated male rats, reproductive organ weights were reduced at this dose level. Reduced pup body weights were observed at this dose level.