Registration Dossier

Administrative data

Endpoint:
two-generation reproductive toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
12 Jun 2014 - 03 Jun 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)
Version / remarks:
adopted in 2001
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.3800 (Reproduction and Fertility Effects)
Version / remarks:
adopted in 1998
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries, Test Data for Registration of Agricultural Chemicals, 12 Nohsan No 8147
Version / remarks:
adopted in 2000
GLP compliance:
yes (incl. certificate)
Remarks:
Department of Health, United Kingdom
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Species:
rat
Strain:
other: RCCHanTM; WIST rat.
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan (UK) Ltd.
- Age at study initiation: (P) 40 - 46 days; (F1) 28 ± 2 days
- Weight at study initiation: (P) 135 - 197 g (males); 112 - 152 g (females); (F1) Males: 77 - 84 g; Females: 72 - 78 g
- Housing: Pre-pairing, males after pairing to termination and females after weaning to termination: groups up to 4 animals per sex per cage in polycarbonate cages with a stainless steel mesh lid and softwood based bark-free fibre bedding; Pairing: 1 male and 1 female in grid bottomed cages above absorbent paper with a stainless steel mesh lid; Females after mating: individually in polycarbonate cages with a stainless steel mesh lid and softwood based bark-free fibre bedding; Females during littering: one female and litter in polycarbonate cages with a stainless steel mesh lid and softwood based bark-free fibre bedding; an aspen chew block and a plastic shelter were provided to each cage except during pairing and lactation
- Diet: SDS VRF1 powdered diet, ad libitum
- Water: potable water from the public supply, ad libitum
- Acclimation period: 18 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 23
- Humidity (%): 40 - 70
- Photoperiod (hrs dark / hrs light): 12 / 12

Administration / exposure

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on exposure:
DIET PREPARATION
- Rate of preparation of diet: weekly
- Mixing appropriate amounts with: SDS VRF1 powdered diet
- Storage temperature of food: ambient (nominally 21°C)
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: up to two weeks
- Proof of pregnancy: ejected copulation plugs in cage tray and sperm in vaginal smear referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged: individual in solid polycarbonate bottom cages with a stainless stell mesh lid with softwood based bark-free fibre bedding and an aspen chew block and plastic shelter as enrichment
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Formulations prepared at nominal concentrations of 150 and 18000 ppm (168 and 20160 ppm formulated concentrations of material as supplied, respectively) were evaluated for stability and homogeneity using HPLC-UV analysis in a previous study. Stability and homogeneity after storage for up to 22 days at either ambient temperature (nominally 21°C) or frozen (nominally -20°C) were confirmed. For concentration analysis via HPLC-UV analysis during the present study, samples of the following formulations were collected: F0 generation (Week 1), F0 gestation, F0-1 generation (final week in lactation), F1 pairing, F1-F2 generation (final week in lactation), F2 maturation (final study week). The mean concentrations of the test substance in test formulations were between -7.7 and +4.7% of nominal values which is within applied limits +10%/-15% of nominal concentrations, confirming accurate formulation.
Duration of treatment / exposure:
approximately 20 weeks (P and F1 generations; males: ten weeks before pairing until termination; females: ten weeks before pairing, throughout pairing, gestation and lactation)
approximately 8 weeks (F2 generation; from weaning until termination on Day 70 of age)
Frequency of treatment:
continously (via diet)
Details on study schedule:
- Selection of parents from F1 generation when pups were 20 days of age.
Doses / concentrationsopen allclose all
Dose / conc.:
300 ppm
Remarks:
during Weeks 1 - 10:
equivalent to 22 and 25 mg/kg bw/day in males and females of the P0 generation, respectively and equivalent to 28 and 30 mg/kg bw/day in males and females of the P1 generation, respectively
Dose / conc.:
600 ppm
Remarks:
during Weeks 1 - 10:
equivalent to 44 and 51 mg/kg bw/day in males and females of the P0 generation, respectively and equivalent to 57 and 63 mg/kg bw/day in males and females of the P1 generation, respectively
Dose / conc.:
2 700 ppm
Remarks:
during Weeks 1 - 10:
equivalent to 196 and 224 mg/kg bw/day in males and females of the P0 generation, respectively and equivalent to 253 and 266 mg/kg bw/day in males and females of the P1 generation, respectively
Dose / conc.:
12 000 ppm
Remarks:
during Weeks 1 - 10:
equivalent to 896 and 1032 mg/kg bw/day in males and females of the P0 generation, respectively and equivalent to 1138 and 1218 mg/kg bw/day in males and females of the P1 generation, respectively
Dose / conc.:
300 ppm
Remarks:
during gestation:
equivalent to 22 and 23 mg/kg bw/day in females of the P0 and the P1 generation, respectively
Dose / conc.:
600 ppm
Remarks:
during gestation:
equivalent to 43 and 47 mg/kg bw/day in females of the P0 and the P1 generation, respectively
Dose / conc.:
2 700 ppm
Remarks:
during gestation:
equivalent to 197 and 211 mg/kg bw/day in females of the P0 and the P1 generation, respectively
Dose / conc.:
12 000 ppm
Remarks:
during gestation:
equivalent to 875 and 1000 mg/kg bw/day in females of the P0 and the P1 generation, respectively
Dose / conc.:
150 ppm
Remarks:
during lactation:
equivalent to 23 and 23 mg/kg bw/day in females of the P0 and the P1 generation, respectively
Dose / conc.:
300 ppm
Remarks:
during lactation:
equivalent to 47 and 47 mg/kg bw/day in females of the P0 and the P1 generation, respectively
Dose / conc.:
1 350 ppm
Remarks:
during lactation:
equivalent to 211 and 215 mg/kg bw/day in females of the P0 and the P1 generation, respectively
Dose / conc.:
6 000 ppm
Remarks:
during lactation:
equivalent to 890 and 947 mg/kg bw/day in females of the P0 and the P1 generation, respectively
No. of animals per sex per dose:
24
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale: The highest dose (12000 ppm) was selected based on a preliminary study of reproduction performance in the rat where treatment-related findings at 15000 ppm consisted of a slightly low body weight gain during late gestation in the F0 parent females compared with the controls together with a slight loss in body weight during late lactation. In the offspring at the high dose level, body weight gain of both male and females was reduced. For animals selected to form the P1 generation, body weight gain continued to be statistically significantly reduced associated with a delay in sexual maturation. The high dose level of 12000 ppm on the two generation reproduction study was expected to approximate to a limit dose of 1000 mg/kg/day during the F0 pre-mating phase of the study. A high intermediate dose level of 2700 ppm provided an approximate 4.5 fold factor to the highest dose level. The selected low intermediate dose of 600 ppm was anticipated to be a NO(A)EL for both parents and offspring. In the event of toxicity observed at 600 ppm, a dose level of 300 ppm was expected to provide a clear NO(A)EL. During the lactation phase the dietary concentration for females was lowered by 50% to 6000, 1350, 300 and 150 ppm so that the achieved dose for the P0/P1 females in the high dose group does not increase markedly above the limit dose of 1000 mg/kg/day as the maternal food consumption increased.
- Rationale for animal assignment:
P0 generation: Animals were allocated to the P0 generation of the study by sex. Animals showing signs of ill health were excluded. Animals at the extreme of the weight range or litters showing large variation in individual weights were not selected if alternatives were available. The method of allocation of animals to the P0 generation ensured that no more than one offspring of each sex from each litter was present in each group.
P1/F2 generation: The offspring with the lowest within-litter identification per sex from each selected litter will be selected to form the P1/F2 generations, after exclusion of grossly atypical animals. Where possible, either two males and two females or one male and one female were selected from each selected litter. If more were required, offspring were taken from randomly selected litters from each group.

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily
- Cage side observations included: mortality, ill-health, reaction to treatment

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly and on Days 0, 5, 12, 18 and 20 after mating and on Days 1, 7, 14 and 21 of lactation for P0 and P1 animals

BODY WEIGHT: Yes
- Time schedule for examinations: P0 males were weighed on the day of treatment, weekly thereafter and at necropsy. P0 females were weighed on the day of treatment, weekly thereafter until mating, on Days 0, 7, 14 and 20 after mating and on Days 1, 4, 7, 14, 21, 25 and 28 post-partum. P1, F1 and F2 animals were weighed at the same frequency P0 animals following selection (nominally 4 weeks of age).

FOOD CONSUMPTION AND COMPOUND INTAKE:
The weight of food supplied to each cage, that remaining and an estimate of any spilled was recorded weekly until paired for mating or termination for P0 males and females. From these records the mean weekly consumption per animal (g/animal/week) was calculated for each cage. The different weights were also recorded for P0 females on Days 0-6, 7-13 and 14-19 after mating and Days 1-3, 4-6, 7-13 and 14-20 of lactation. From these records the mean daily consumption (g/animal/day) was calculated for each animal. For P1/F1/F2 animals the weight of food supplied to each cage, that remaining and an estimate of any spilled was recorded at the same frequency as P0 animals following selection, at approximately 4 weeks of age.

WATER CONSUMPTION: No
Oestrous cyclicity (parental animals):
Smears were taken daily for 22 days before pairing, using cotton swabs moistened with saline. The smears were subsequently examined to establish the duration and regularity of the oestrous cycle. After pairing with the male, daily smearing was continued using pipette lavage, until evidence of mating was observed. For 4 days before scheduled termination, Days 25 to 28 post-partum for P0 and P1 females and Days 67 to 70 of age for F1 and F2 females, daily vaginal smears were taken and used to determine the stage of the oestrous cycle at termination.
Sperm parameters (parental animals):
Parameters examined in P/F1 male parental generations:
testis weight, epididymis weight, sperm motility, sperm morphology, sperm count in epididymides, other: homogenisation-resistant spermatids count
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes
- Maximum of 8 pups/litter (4/sex/litter wherever possible); excess pups were killed and discarded.

PARAMETERS EXAMINED
The following parameters were examined in F1 / F2 offspring:
number and sex of pups, anogenital distance (AGD), postnatal mortality, other: clinical signs, sex ratio, individual offspring body weights

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death was not determined for pups born or found dead; an assessment for the presence of milk in the stomach was included

ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY: No

ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY: No
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: all surviving animals when the majority of litters had weaned after 18 weeks of treatment for the P0 males or 18 weeks of the F1 generation for the P1 males
- Maternal animals: all surviving animals on Day 28 post-partum

GROSS NECROPSY
- Gross necropsy consisted of a complete macroscopic examination. For all females of the parental generations the number of implantation sites was recorded.

HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table 1 were prepared for microscopic examination and weighed, respectively.
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring not selected as parental animals and all F2 offspring were sacrificed at 21 days of age.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:

GROSS NECROPSY
- Gross necropsy consisted of a complete macroscopic examination.

HISTOPATHOLOGY / ORGAN WEIGTHS
The tissues indicated in Table 2 were prepared for microscopic examination and weighed, respectively.
Statistics:
For body weight, food consumption, litter size and survival indices, sexual maturation and organ weight data, a parametric analysis was performed if Bartlett's test for variance homogeneity was not significant (1% level). The F1 approximate test was applied. If the F1 approximate test for monotonicity of dose-response was not significant (1% level), Williams' test for a monotonic trend was applied. If the F1 approximate test was significant, Dunnett's test was performed instead. A non-parametric analysis was performed if Bartlett's test was still significant at the 1% level following both logarithmic and square-root transformations. The H1 approximate test, the non-parametric equivalent of the F1 test described above, was applied. If the H1 approximate test for monotonicity of dose-response was not significant at the 1% level, Shirley's test for a monotonic trend was applied. If the H1 approximate test was significant, Steel's test was performed instead. For litter size and survival indices and sexual maturation, if 75% of the data (across all groups) were the same value, for example c, Fisher’s Exact tests were performed (pairwise comparison). Sex ratio were analysed by generalised mixed linear model with binomial errors, a logit link function and litter as a random effect. Each treated group was compared to control using a Wald chi-square test. For sex ratio, the numerator was number of males, the denominator was number of live fetuses. For gestation length an exact two-tailed Linear-by-linear test, with equally spaced scores, was applied to all groups. If the test was statistically significant (p<0.05), the highest dose group was excluded and the test re-applied. This ‘step-down’ process was repeated until the test was no longer statistically significant (p≥0.05). If the exact version of the Linear-by-linear test could not be calculated (due to the size of the table containing the data), the asymptotic version was used instead.
Reproductive indices:
Percentage mating = (number animals mating / animals paired) x 100
Conception rate (%) = (number animals achieving pregnancy / animals mated) x 100
Fertility index (%) = (number animals achieving pregnancy / animals pairing) x 100
Gestation index (%) = (number live litters born / number pregnant) x 100
Post-implantation survival index (%) = (total number of offspring born / total number of uterine implantation sites) x 100
Percentage males = (number of males in litter / total number of offspring in litter) x 100
Offspring viability indices:
Live birth index (%) = (number of live offspring on Day 1 after littering / total number of offspring born) x 100
Viability index (%) = (number of live offspring on Day 4 before culling / number of live offspring on Day 1 after littering) x 100
Lactation index (%) = (number of live offspring on Day 21 after littering / number of live on Day 4 after culling) x 100

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not examined
Mortality:
mortality observed, non-treatment-related
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
FEMALES
12000 ppm: decreased mean body weight gain when compared to controls during the first and second week of gestation (Days 0 - 7 and 7 - 14); increased body weight gain when compared to controls from Day 1 - 4 of lactation (both adverse during Days 0 - 7)
2700, 600 and 300 ppm: decreased mean body weight gain when compared to controls during the second week of gestation (Days 7 - 14)

(see table 3)
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
MALES
12000, 2700 and 600 ppm: decreased food consumption when compared with controls during Week 1 (adverse at the highest dose)
12000 and 2700 ppm: decreased food consumption when compared with controls during Week 2 (adverse at the highest dose)
12000, 2700, 600 and 300 ppm: decreased food consumption when compared with controls during Weeks 5,7 and 8 (adverse at the highest dose)

FEMALES
12000, 2700, 600 and 300 ppm: decreased food consumption when compared with controls during Week 1; decreased food consumption when compared with controls during the second week of gestation (adverse at the highest dose)

(see table 4)
Food efficiency:
no effects observed
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
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
effects observed, non-treatment-related
Other effects:
no effects observed

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
no effects observed
Reproductive performance:
effects observed, treatment-related
Description (incidence and severity):
FEMALES
12000 ppm: significant shift towards shorter gestation periods with the percentage exhibiting a gestation length of 22 days exceeding the historical control data range (non-adverse)
(see table 5)

Pre-coital interval, mating performance, fertility and gestation index were unaffected by treatment

Details on results (P0)

For details on results including tables see the attached document.

Effect levels (P0)

open allclose all
Key result
Dose descriptor:
NOAEL
Remarks:
systemic toxicity
Effect level:
2 700 ppm
Based on:
test mat.
Remarks:
equivalent to 196 and 224 mg/kg bw/day in males and females, respectively
Sex:
male/female
Basis for effect level:
body weight and weight gain
food consumption and compound intake
Key result
Dose descriptor:
NOAEL
Remarks:
reproductive toxicity
Effect level:
>= 12 000 ppm
Based on:
test mat.
Remarks:
equivalent to 896 and 1032 mg/kg bw/day for males and females, respectively
Sex:
male/female
Basis for effect level:
other: highest dose tested

Target system / organ toxicity (P0)

Key result
Critical effects observed:
no

Results: P1 (second parental generation)

General toxicity (P1)

Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not examined
Mortality:
mortality observed, non-treatment-related
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
MALES
12000, 2700, 600 and 300 ppm: decreased mean body weight at the beginning of the P1 generation (Week 0) and during Weeks 1, 2, 3, 4, 5 and 6 (adverse at the highest dose)

FEMALES
12000, 2700, 600 and 300 ppm: decreased mean body weight at the beginning of the P1 generation (Week 0) and during all weeks before pairing; decreased weight gain between Weeks 0 - 10 before pairing, decreased mean body weights during gestation and lactation, increased weight gain during Days 1 - 4 of lactation (all effects adverse at the highest dose)

(see table 6)
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
MALES
12000 and 2700 ppm: decreased food consumption during Week 1 (adverse at the highest dose)
600 ppm: increased food consumption during Week 2
12000, 2700, 600 and 300 ppm: decreased food consumption during Weeks 3 and 4 (adverse at the highest dose)

FEMALES
12000 and 2700 ppm: decreased food consumption during Week 1 (adverse at the highest dose)
600 ppm: increased food consumption during Week 2
12000, 2700, 600 and 300 ppm: decreased food consumption during Week 4 (adverse at the highest dose)

(see table 7)
Food efficiency:
no effects observed
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
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Other effects:
no effects observed
Details on results:
For details on results including tables see the attached document.

Reproductive function / performance (P1)

Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
no effects observed
Reproductive performance:
effects observed, treatment-related
Description (incidence and severity):
Pre-coital interval, mating performance and fertility were unaffected by treatment. Assessment of vaginal cytology prior to termination did not reveal any differences across the groups. Assessment of ovarian follicle counts of control females and females at the highest dose (12000 ppm and 6000 ppm during lactation phase) did not reveal any difference.
There was a slight shift in gestation length with a higher percentage of females at 12000 ppm showing a shorter gestation length compared with controls, although, the gestation length for all females at 12000 ppm was within the normal range of 22 to 23.5 days. The shift did not attain statistical significance, however the percentage of females showing a 22-day gestation length exceeded the historical control data range, whilst the percentage of females showing a 22.5 and 23-day gestation length was below the historical control data range. It should be noted however, that the concurrent control data was also outside the historical control data range.

Effect levels (P1)

open allclose all
Key result
Dose descriptor:
NOAEL
Remarks:
systemic toxicity
Effect level:
2 700 ppm
Based on:
test mat.
Remarks:
equivalent to 253 and 266 mg/kg bw/day for males and females, respectively
Sex:
male/female
Basis for effect level:
body weight and weight gain
food consumption and compound intake
Key result
Dose descriptor:
NOAEL
Remarks:
reproductive toxicity
Effect level:
>= 12 000 ppm
Based on:
test mat.
Remarks:
equivalent to 1138 and 1218 mg/kg bw/day for males and females, respectively
Sex:
male/female
Basis for effect level:
other: highest dose tested

Target system / organ toxicity (P1)

Key result
Critical effects observed:
no

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not examined
Mortality / viability:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
MALES
12000/6000 ppm (parental animal before lactation phase/during lactation phase): decreased offspring body weight gain when compared with controls from Day 7 of age up to weaning on day 21 of age
12000, 2700, 600 and 300 ppm (treatment following weaning at Days 21 - 25 in selected animals): decreased offspring body weight gain when compared to controls

FEMALES
12000/6000 ppm (parental animal before lactation phase/during lactation phase): decreased offspring body weight gain when compared with controls from Day 7 of age up to weaning on day 21 of age
2700/1350, 600/300 and 300/150 ppm (parental animal before lactation phase/during lactation phase): decreased offspring body weight gain when compared with controls from Day 7 to 14 of age

(see table 8)

Body weight and weight changes during the 10 week pre-mating period of the F1 generation becoming P1 generation are shown in section "Results: P1 generation".
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Food consumption during the 10 week pre-mating period of the F1 generation becoming P1 generation is shown in section "Results: P1 generation".
Food efficiency:
no effects observed
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:
effects observed, non-treatment-related
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Gross pathological findings:
no effects observed
Histopathological findings:
no effects observed
Other effects:
no effects observed

Developmental neurotoxicity (F1)

Behaviour (functional findings):
not examined

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
not examined

Details on results (F1)

For details on results including tables see the attached document.

Effect levels (F1)

open allclose all
Key result
Dose descriptor:
NOAEL
Remarks:
systemic toxicity
Generation:
F1
Effect level:
2 700 ppm
Based on:
test mat.
Remarks:
equivalent to 253 and 266 mg/kg bw/day in males and females, respectively
Sex:
male/female
Basis for effect level:
body weight and weight gain
food consumption and compound intake
Key result
Dose descriptor:
NOAEL
Remarks:
reproductive toxicity
Generation:
F1
Effect level:
>= 12 000 ppm
Based on:
test mat.
Remarks:
equivalent to 1138 and 1218 mg/kg bw/day in males and females, respectively
Sex:
male/female
Basis for effect level:
other: highest dose tested

Target system / organ toxicity (F1)

Key result
Critical effects observed:
no

Results: F2 generation

General toxicity (F2)

Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not examined
Mortality / viability:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
MALES
12000/6000 ppm (parental animal before lactation phase/during lactation phase): decreased offspring weight gain when compared with controls from Day 14 to 21 of age; decreased absolute weights from Day 14 to 25
12000 ppm: decreased absolute body weights when compared with controls on Weeks 0, 1, 2, 4 and 5 (adverse)

FEMALES
12000/6000 ppm (parental animal before lactation phase/during lactation phase): decreased offspring weight gain when compared with controls from Day 14 to 21 of age; decreased absolute weights from Day 21 to 25
12000 ppm: decreased absolute body weights when compared with controls on Weeks 1, 4 and 5 (adverse)

(see tables 12 and 13)
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
MALES
12000 and 2700 ppm: decreased food consumption during Week 3

(see table 14)
Food efficiency:
no effects observed
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:
effects observed, non-treatment-related
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
MALES
12000/6000 ppm (parental animal before lactation phase/during lactation phase): increased brain weight relative to body weight (see table 16)
12000 ppm (selected animals terminated on Day 70 of age): decreased absolute and body weight relative ovarian weights (see table 17)
Gross pathological findings:
no effects observed
Histopathological findings:
not examined
Other effects:
effects observed, non-treatment-related

Developmental neurotoxicity (F2)

Behaviour (functional findings):
not examined

Developmental immunotoxicity (F2)

Developmental immunotoxicity:
not examined

Details on results (F2)

For details on results including tables see the attached document.

Effect levels (F2)

open allclose all
Key result
Dose descriptor:
NOAEL
Remarks:
systemic toxicity
Generation:
F2
Effect level:
2 700 ppm
Based on:
test mat.
Remarks:
equivalent to 307 and 312 mg/kg bw/day for males and females, respectively
Sex:
male/female
Basis for effect level:
sexual maturation
body weight and weight gain
food consumption and compound intake
Key result
Dose descriptor:
NOAEL
Remarks:
reproductive toxicity
Generation:
F2
Effect level:
>= 12 000 ppm
Based on:
test mat.
Remarks:
equivalent to 1361 and 1392 mg/kg bw/day for males and females, respectively
Sex:
male/female
Basis for effect level:
other: highest dose tested

Target system / organ toxicity (F2)

Key result
Critical effects observed:
no

Overall reproductive toxicity

Key result
Reproductive effects observed:
no

Any other information on results incl. tables

For details on results including tables see the attached document.


 

Applicant's summary and conclusion