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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
Study period:
March to August 1992
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: well documented study performed according to OECD guideline

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1993
Report Date:
1993

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 415 [One-Generation Reproduction Toxicity Study (before 9 October 2017)]
Version / remarks:
adopted May 26, 1983
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
- Name of test material (as cited in study report): Marlotherm S
- Substance type: pure active substance
- Physical state: yellow liquid
- Lot/batch No.: 200 l barrel from August 21, 1991
- Stability under test conditions: unlimited
- Storage condition of test material: no data
Specific details on test material used for the study:
The purity of the material used for the test was lower than the purity of the actual material. It contained ca. 26% of isomers of dimethyl tribenzenes. However, based on comparable basic structure of the impurities and the assumption that any toxic effects would increase with higher molecular weight and lipophilicity, we regard that the studies conducted with the lower purity material being "worst case" and can be used for the evaluation of the actual product.

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Wiga GmbH, Sulzfeld, Germany
- Age at study initiation: 6 wks (males) and 8 wks (females)
- Weight at study initiation: Males: 208-261 g; Females: 188-243 g
- Fasting period before study: no
- Housing: single in Macrolon cages (except during mating and lactation)
- Diet: "Ssniff R" pelleted diet produced by Ssniff Spezialitätendiäten GmbH, Soest, Germany; ad libitum
- Water: tap water; ad libitum
- Acclimation period: 12 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.5 +/- 1.5
- Humidity (%): 50-70
- Air changes (per hr): 16
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: 0.5 % Sodium Carboxymethylcellulose
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
All dosing solutions were prepared freshly each day by means of an ultraturrax. The emulsions were permanently stirred in the animal room prior to administration in order to maintain homogeneity.


VEHICLE
- Justification for use and choice of vehicle (if other than water): no data
- Concentration in vehicle: according to the dose levels 0, 2, 12 and 72 g/l, respectively
- Amount of vehicle (if gavage): 10 ml/kg bw
- Lot/batch no. (if required): 109 F 0361 (supplied by Sigma Deisenhofen)
- Purity: 0.5 % aqueous solution
Details on mating procedure:
- M/F ratio per cage: 1/1
- Length of cohabitation: 15 hours
- Proof of pregnancy: vaginal plug and/or sperm in vaginal smear referred to as day 0 of pregnancy
- After 21 days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: no
- After successful mating each pregnant female was caged (how): singly in Macrolon cages
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Dichloromethane extracts of the dose suspensions were analysed by photometry.
Additionally the stability of test substance in dose suspensions were analysed over an 24 hour interval.
Duration of treatment / exposure:
Males were treated for 10 weeks prior to mating and throughout mating up to sacrifice.
All females were treated for 2 weeks prior to mating and throughout mating, gestation and lactation up to weaning on, or shortly after day 28 post partum. Treatment ceased on the day prior to sacrifice.
Frequency of treatment:
once daily
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
20 mg/kg bw
Basis:
actual ingested
Remarks:
Doses / Concentrations:
120 mg/kg bw
Basis:
actual ingested
Remarks:
Doses / Concentrations:
720 mg/kg bw
Basis:
actual ingested
No. of animals per sex per dose:
24
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale:
A preliminary experiment was carried out in pregnant rats with dose levels of 100, 500 and 1000 mg/kg bw. During the course of this study a clear effect upon weight development was noted during the first week of treatment of pregnant rats at all dose levels. Food consumption and food conversion ratios were also effected.
During a 3-month oral toxicity study in rats increased liver and kidney weights were found at dose levels of 100 and 500 mg/kg. The high dose of 720 mg/kg was selected to obtain clear toxic effects according to the recommendation of the OECD guideline. The low and mid doses were chosen in order to obtain information about possible dose-related effects and in order to establish a clear "no observable adverse effect level".
Positive control:
no

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: No data


DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily observations. Viability was checked twice daily. Dose response examinations were carried out at appropriate intervals after administration of the test article.


BODY WEIGHT: Yes
- Time schedule for examinations:
Males: Males were weighed prior to commencement of treatment, at weekly intervals during pre-mating treatment and after the mating period and at terminal sacrifice.

Females: All females were weighed prior to commencement of treatment and at weekly intervals during the pre-mating treatment phase. Weights were also recorded on days 0, 7, 14 and 20 of gestation; dams that littered were weighed on day 0 or 1 and on days 4, 7, 14, 21 and 28 post partum.
For non-mated females, weekly body weight determinations were continued until sacrifice, beginning on the first day after the mating period.

During mating, both males and females were weighed at least weekly for dose adjustment purposes; these body weights were not recorded. During gestation and lactation females were weighed daily for dose adjustment.


FOOD CONSUMPTION: Yes
- Time schedule for examinations:
Males: Food intake was determined at weekly intervals during pre-mating treatment until terminal sacrifice.

Females: Food intake was determined at weekly intervals during the pre-mating treatment phase. Food intake was also recorded on days 7, 14 and 20 of gestation; dams that littered were weighed on day 0 or 1 and on days 4, 7, 14, 21 and 28 post partum.
For non-mated females, weekly body weight determinations were continued until sacrifice, beginning on the first day after the mating period.

Food consumption was not determined during mating.
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no

PARAMETERS EXAMINED
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities, age of developmental stage (pinna folding, hair growth, eye opening, upper incisor eruption), testis descent, surface righting reflex, startle reflex, air righting reflex, pupil reflex, ophthalmoscopy, special clinical examinations (modified Irwin Screening); between days 25 and 28 post partum behavioural examinations were carried out in at least 10 male and 10 female pups of each group (rotating rod test, inclined plane test, open field test)


GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals at the end of the mating period
- Parent females: All surviving animals on, or shortly after, day 28 of lactation, or after death of the whole litter
Females that failed to mate successfully or which were considered to have mated successfully but failed to produce a viable litter were sacrified approx. 26 days after the last day of the mating period.

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the internal viscera.
Females that failed to mate successfully were examined macroscopically for organ changes with special attention to the organs of the reproduction tract.

HISTOPATHOLOGY/ORGAN WEIGHTS
Males: liver, testes with epididymides, seminal vesicles with coagulating gland, prostate, combined weights of kidneys (only in control and high dose group animals). These organs, together with the pituitary gland and any tissues showing severe changes, were preserved.
Females: Weights of liver and combined kidneys were determined only in control and high dose group. Ovaries, uterus, cervix, vagina, pituitary gland and tissues showing severe changes, were preserved.
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring were sacrificed at 28 days of age.
- These animals were subjected to postmortem examinations as follows:


GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations


HISTOPATHOLOGY / ORGAN WEIGTHS
Where possible, two male and two female pups were selected from each litter in order to determine the weight of brain, heart, liver, kidneys and spleen except the weight of liver in females of low and mid dose group (due to operational reasons).
Tissue showing severe changes were preserved.
Statistics:
Body weight changes, food consumption, pup weights and the number of implantations and offspring were subjected to analyses of variance with a subsequent multiple range test, or, if indicated, group mean values were compared by the "Kruskal-Wallis test" and "Mann-Whitney-U-test".
The quotient (weight changes/food consumtion) x 100 was calculated for each determination phase (food conversion ratio)
Organ weights were evaluated as both absolute and relative weights (% body weight). Dose groups were compared to the control group by the Dunnett test (two-tailed), modified according to Kramer.
Sex distribution, gestation length, the number of days until successful mating, physical and reflex development and behaviour in the rotating rod, inclined plane and open field tests were analysed by "Kruskal-Wallis test" and "Mann-Whitney-U-test".
Indices were compared by an appropriate statistical method, if indicated.
Where appropriate, the basic unit for all parameters was the litter.

References:
- Winer, B. J. (1971) Statistical principles in experimental design, International student edition, McGraw Hill Kogakusha Ltd.
- Snedecor, G. W. and W. G. Cochran (1967) Statistical methods. 6th ed. Iowa State University Press Ames, Iowa.
- Dunnett, C. W. (1955) A multiple comparison procedure for comparing several treatments with a control. J. Am. Stat. Assoc. 50, 1096-1121.
- Kramer, C. Y. (1956) Extension of multiple range test to group means with unequal numbers of replications. Biometrics 12, 307-310.
Reproductive indices:
Mating Index = 100 x No. of females in which mating was confirmed/No. of paired females
Fertility Index= 100 x No. of pregnant females/No. of paired females
Conception Rate= 100 x pregnant females/No. of females in which mating was confirmed
Abortion rate = 100 x No. of dams showing abortion/No. of pregnant dams
Gestation Index = 100 x No. of litters with live fetuses/No. of pregnant females
Pre-birth Loss Index = 100 x (No. of implantations - total litter size at birth)/No. of implantations
Pup Loss Index (at birth)= 100 x (total litter size at birth - live litter size at birth)/Total litter size at birth
Cumulative Pup Loss Index (Day x)= 100 x (total litter size at birth - live litter size on day x)/Total litter size at birth
Sex Ratio= 100 x No. of live male pups per litter/No. of live pups per litter

Results and discussion

Results: P0 (first parental animals)

General toxicity (P0)

Clinical signs:
effects observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
not examined
Other effects:
no effects observed

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)

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
No treatment-related clinical findings were noted in males and females of all dose groups except salivation which was observed in males in the high dose group during pre-mating only in two animals and nearly all animals after the mating period. Salivation was observed in females during gestation from day 8 onwards in high dose group and during lactation in mid and high dose group with dose-related increase in incidence. Salivation is considered to be a local effect of the test article.
Two males of the control group showed dermatopathia on the neck from weeks 2 and 3 onwards which was treated with dexpanthenol ointment from week 5 until week 7 or 8, respectivlely. The the animals displayed alopecia only.
One female of the high dose group appeared emaciated during the last weeks of lactation.
One male of the mid dose group was found dead at the end of week 13 of treatment. This spontaneous death is considered to be without toxicological importance. One female of high dose group was found dead on day 24 of lactation.

BODY WEIGHT (PARENTAL ANIMALS)
Weight gain rates of high dose males were significantly reduced during the entire treatment period (see table 1). In females during the pre-mating period reduced body weight gain was observed in mid and high dose group but attained significance only in the high dose group. During gestation decrease in weight gain of high dose females was observed between days 0 and 7 and 14 and 20 and over the whole period of gestation. Between beginning and end of lactation weight gain of high dose females was significantly increased relative to control females. They showed a smaller weight loss from days 14 to 21 and 21 to 28 compared to controls resulting in higher absolute body weights of high dose females compared to control. The increased weight gain was probably a compensation of decreased body weight at delivery as the mean body weigth of the high dose group females were decreased at the start of lactation and only slightly increased on day 28 compared to control females. (see table 2 and 3)

FOOD CONSUMPTION AND FOOD CONVERSION RATIO (PARENTAL ANIMALS)
No significant changes in food consumption was found in males and females during pre-mating treatment. During gestation food consumption was significantly reduced in high dose females between days 1 and 7 and over the entire period of gestation. During the lactation period mean food consumption values were slighly decreased in high dose females on days 14, 21 and 28 of lactation but with a high standard deviation. In addition the pups also eat the provided food particularly in the last part of lactation and the number of pups per dam was decreased in high dose group.
Food conversion ratio of males showed only slighly decreased mean values in high dose group in weeks 3, 4, 6, 7, and 8 of treatment. This corresponds to the reduced weight gain in this dose group. In females the food conversion ratio was decreased in high dose females during week 1 of pre-mating treatment period and slightly on day 20 of gestation.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
Mating index, mating performance, fertility index and conception rate did not reveal any differences attributable to the treatment. The fertility index and conception rate of the high dose group were decreased but both were within the historical range of historical data and the differences were not statistically significant.
No females showed signs of abortion or premature delivery. Gestation indices did not reveal treatment-related differences. The decreased value of the high dose group was due to one animal only. No significant differences were found for gestation lenghts. No instances of prolonged parturition or dystocia. No significant differences were found either in the total number of implantations or in the distribution between the right and left uterine horns. Pre-birth loss indices did not differ significantly between groups and no-doserelationship was evident.

ORGAN WEIGHTS (PARENTAL ANIMALS)
The relative and absolute liver weights were significantly increased in males of all dose groups with a clear dose-relationship and in females only in the high dose group. Absolute and relative kidney weights were significantly increased in males of the high dose group. In females absolute kidney weights were significantly increased in the high dose group. However since the body weights of these females were higher the relative weights did not reveal a significant difference. Thus, an effect on the weights of kidneys cannot be determined for the females. (see table 4)
With respect to the reproductive organs, relative prostate weights were significantly increased in high dose males. Since mating performance and fertility did not reaveal any treatment-related differences, this finding may be of minor toxicological importance. The weights of testes and seminal vesicles did not show any significant differences. (see table 4)

GROSS PATHOLOGY (PARENTAL ANIMALS)
No distinct treatment-related findings were found during necropsy of both males and females.

Effect levels (P0)

open allclose all
Key result
Dose descriptor:
NOAEL
Effect level:
120 mg/kg bw/day (actual dose received)
Sex:
male
Basis for effect level:
other: based on body weight; food consumption; organ weights; liver weight effects appear to be an adaptive response rather than a toxic effect;
Key result
Dose descriptor:
NOAEL
Effect level:
120 mg/kg bw/day (actual dose received)
Sex:
female
Basis for effect level:
other: body weight; food consumption; organ weights
Dose descriptor:
NOAEL
Remarks:
reproductive toxicity
Effect level:
ca. 720 mg/kg bw/day (actual dose received)
Sex:
male/female
Basis for effect level:
other: absence of effects on reproductive functions

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Mortality / viability:
mortality observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
no effects observed
Histopathological findings:
not examined

Details on results (F1)

LITTER SIZE
The litter size was significantly reduced in the high dose group on days 4, 7, 14, 21 and 28 post partum which was predominantly caused by the increased pup loss after birth. (see table 5)

SEX DISTRIBUTION, SEX RATIO (OFFSPRING)
The sex ratio did not reveal important differences. The percentage of males showed only slight intergroup variation, which was considered to be within the physiological range.

VIABILIY INDEX (OFFSPRING)
The pup loss index at birth was not affected by treatment in all test groups. Correspondingly, the number of litters with dead pups at birth was very similar between all groups.
Cumulative pup loss indices were increased in the high dose group from day 7 post partum onwards. (see table 6)
The cumulative pup loss index of the mid dose group also seemed to be increased. But an evaluation of the individual mid dose group litters affected showed that the increased cumulative pup loss index was predominantly caused by one litter, in which 15 of 16 pups were found dead. Additionally with respect to the number of litters with dead pups, an increase in the mid dose group cannot be determined.

CLINICAL SIGNS (OFFSPRING)
No clear indication of treatment effect on the appearance and general condition of offspring.

BODY WEIGHT (OFFSPRING)
Mean group body weights of both male and female pup were significantly reduced in the high dose group at birth and on days 4, 7, 14, 21 and 28 post partum. (see table 7)

ORGAN WEIGHTS (OFFSPRING)
Relative weights of spleen did not reveal any significant differences between the groups. Relative heart weights were significantly increased in high dose group males and females. Decreased relative heart weights in males of the the low dose group is considered to be not treatment-related, since relative heart weights in mid dose group are lower (no clear dose-relationship) and no increase of relative heart weights was found in females of the low dose group.
Relative kidney weights and relative brain weights were significantly increased in high dose males and females. Decreased relative brain weights in males and females of the low dose group is considered to be not treatment-related as the brain weights of the mid dose group were close to those of the control (no clear dose-relationship). Also the significant difference in relative liver weights of the low dose females is considered to be incidental since no dose-relationship existed. (see table 8)

GROSS PATHOLOGY (OFFSPRING)
Necropsy of pups found dead during pre-weaning development did not reveal any specific findings which point to a manifestation of parental treatment-effect.
Macroscopic examinations of pups sacrificed at weaning revealed no changes clearly attributable to the treatment of parents. A diaphragmatic hernia was found in one pup of the high dose group. This finding is believed to be associated with the distinct growth or developmental retardation found in this dose group.

PHYSICAL DEVELOPMENT (OFFSPRING)
No obviously treatment-related retardations were found for the live pups. Group mean values of cumulative frequencies of pups which had attained each specific developmenal stage were comparable between the groups with the exception of hair growth. A slightly delayed commencement of hair growth was observed in the high dose group, as the cumulative frequency on days 5 and 6 was decreased relative to control. Although not statistically significant, this decrease corresponds to the reduction of group mean body weights in the high dose pups.
The occasional significant differences in the low and mid dose group are considered to be incidental.

REFLEX DEVELOPMENT (OFFSPRING)
The onset of the startle reflex was definitely retarded in the high dose group pups; the cumulative frequency of responding pups was significantly decreased relative to control. All pups of the high dose group showed this reflex only on day 18 post partum whereas control pups did on day 15 post partum. This retardation was most probably in corrspondence to the reduced body weight development.
Evaluation of the development of the other reflexes examined did not reveal any treatment-related differences.

OTHER FINDINGS (OFFSPRING)
During ophthalmoscopic examination of pups after day 18 post partum only a few spontaneous findings were ascertained.
The modified Irwin Screening of live pups did not provide evidence of a treatment-related effect.
The inclined plane test showed a significantly decreased clinging ability of high dose group female pups. A slightly but non-significantly decreased clinging ability may also be stated for high dose group male pups. This decrease seems to be in correspondence with the affected weights of the brain and the developmental retardations which were found in high dose group pups.
Results of the open field test did not provide clear evidence of a treatment-related effect. Since no clear dose-relationship is obvious a treatment-related effect cannot be stated.
No significant differences were found during the rotating rod test in both male and female pups.

Effect levels (F1)

Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
720 mg/kg bw/day (actual dose received)
Sex:
male/female
Basis for effect level:
other: all effects observed at the high dose level (e.g. litter size; pup weight;) are linked to the body weight reduction of the pups itself and appear to be secondary to maternal toxicity.

Overall reproductive toxicity

Reproductive effects observed:
not specified

Any other information on results incl. tables

Table 1: Weight changes (parental males; mean values and standard deviations)
Group Control 20 mg/kg 120 mg/kg 720 mg/kg
Weeks 0-15  absolute (g) 383.3 ± 48.3 385.2 ± 50.3 390.5 ± 38.9 346.0 ± 47.0*
relative (%) 100.0 100.5 101.9 90.3
Weeks 5-10 absolute (g) 105.8 ± 20.1 101.2 ± 18.5 101.6 ± 20.5 87.8 ± 23.0*
relative (%) 100.0 95.7 96.1 83.1
Weeks 10-15  absolute (g) 48.4 ± 17.8 49.7 ± 13.9 45.2 ± 20.3 34.7 ± 14.0*
relative (%) 100.0 102.7 93.3 71.6
Weeks 13-15  absolute (g) 26.3 ± 11.0 20.3 ± 10.9 25.1 ± 15.7 19.3 ± 8.4*
relative (%) 100.0 77.5 95.6 73.5
*p < 0.05 (Scheffé test)
Table 2: Weight changes (parental females; mean values and standard deviations)
Group Control 20 mg/kg 120 mg/kg 720 mg/kg
Weeks 0-2 (pre-mating)  absolute (g) 39.1 ± 10.5 37.9 ± 12.2 34.6 ± 7.9 31.4 ± 7.1*
relative (%) 100.0 96.8 88.5 80.3
Days 0-20 (gestation)  absolute (g) 156.7 ± 22.4 150.1 ± 23.4 153.8 ± 23.7 133.7± 19.4*
relative (%) 100.0 95.8 98.2 85.3
Days 0-7 (gestation)  absolute (g) 37.6 ± 7.9 37.9 ± 7.1 38.6± 9.5 34.6 ± 7.0*
relative (%) 100.0 100.8 102.6 91.6
Days 14-20 (gestation)  absolute (g) 82.4 ± 12.2 80.9 ± 19.5 82.3 ± 20.2 70.5 ± 12.8*
relative (%) 100.0 98.2 99.9 85.6
Days 0/1-28 (lactation)  absolute (g) -25.8 ± 19.0 -18.7 ± 35.0 -22.0 ± 19.6 5.8 ± 16.8*
relative (%) 100.0 72.3 85.1 -22.3
Days 14-21 (lactation)  absolute (g) -26.6 ± 20.2 -24.0 ± 17.0 -15.2 ± 17.3 -4.6 ± 13.2
relative (%) 100.0 90.5 57.1 17.1
Days 21-28 (lactation)  absolute (g) -36.7 ± 19.3 -39.7 ± 21.8 -37.5 ± 17.2 -18.5 ± 14.4*
relative (%) 100.0 108.0 102.0 50.3
*p < 0.05 (Dunnett-test)
Table 3: Body weights of parental females during lactation (Mean values and standard deviations)
Group Control 20 mg/kg 120 mg/kg 720 mg/kg
Day 0/1 (lactation) 323.4 ± 25.6 313.7 ± 37.7 329.9 ± 28.6 304.2 ± 18.0
Day 4 (lactation) 338.0 ± 31.0 332.9 ± 22.4 344.0 ± 33.4 311.2 ± 18.1
Day 7 (lactation) 357.4 ± 30.3 351.2 ± 22.7 356.4 ± 28.5 327.2 ± 21.2
Day 14 (lactation) 360.9 ± 29.0 358.7 ± 23.1 360.6 ± 25.0 333.8 ± 23.6
Day 28 (lactation) 297.6 ± 25.6 295.0 ± 16.6 308.0 ± 28.2 311.4 ± 15.6
Table 4: Selected organ weights of parental animals (mean values and standard deviations)
Group Control 20 mg/kg 120 mg/kg 720 mg/kg
Liver weights males 23.2310 ± 3.0147 25.7816 ± 4.1249* 27.8573 ± 2.7969* 29.5665 ± 4.4976*
absolute (g) females 16.0065 ± 1.9983 22.7884 ± 2.7078*
Liver weights males 3.771 ± 0.361 4.137 ± 0.430* 4.456 ± 0.302* 5.089 ± 0.436*
relative (%) females 5.340 ± 0.524 7.395 ± 0.395*
Kidney weights males 4.1578 ± 0.5618 4.8922 ± 0.6641*
absolute (g) females 2.7601 ± 0.2707 3.0071 ± 0.3162*
Kidney weights males 0.675 ± 0.071 0.845 ± 0.087
relative (%) females 0.922 ± 0.079 0.977 ± 0.100
Prostate weights absolute (g) 1.6846 ± 0.1821 1.6622± 0.2078 1.7406 ± 0.2621 1.7579 ± 0.2005
relative (%) 0.2752 ± 0.0347 0.2689 ± 0.0340 0.2801 ± 0.0488 0.3057 ± 0.0419*
*p < 0.05 (Dunnett-test, two-tailed)
Table 5: Litter size; No. of pups (mean values and standard deviations)
Group Control 20 mg/kg 120 mg/kg 720 mg/kg
at birth (total) 15.0 ± 2.8 15.3 ± 2.9 14.4 ± 3.4 13.6 ± 3.6
at birth (live pups) 14.9 ± 2.8 15.2 ± 3.0 14.1 ± 3.3 13.4 ± 3.8
Day 4 14.7 ± 2.8 15.1 ± 3.0 13.4 ± 4.1 12.8 ± 3.8*
Day 7 14.5 ± 2.7 14.9 ± 3.0 13.2 ± 4.1 12.2 ± 4.1*
Day 14 14.5 ± 2.7 14.9 ± 3.0 13.1 ± 4.1 12.1 ± 4.3*
Day 21 14.5 ± 2.7 14.9 ± 3.0 13.1 ± 4.1 11.9 ± 4.3*
Day 28 14.5 ± 2.7 14.9 ± 3.0 13.1 ± 4.1 11.9 ± 4.4*
number of litters 21 23 22
* p (U-test) < 0.05
Table 6: Cumulative pup loss index (%) 
Group Control 20 mg/kg 120 mg/kg 720 mg/kg
Day 4 2.8 1.1 7.0 5.8
Day 7 3.8 2.3 8.2 10.4
Day 14 4.1 2.6 8.5 11.6
Day 21 4.1 2.6 8.5 12.4
Day 28 4.1 2.6 8.5 12.7
Tabe 7: Body weight of offspring (g; mean values and standard deviations)
Group Control 20 mg/kg 120 mg/kg 720 mg/kg
males
Day 0/1 6.75 ± 0.48 6.44 ± 5.6 6.78 ± 0.72 6.19 ± 0.53*
Day 4 10.02 ± 1.08 9.64 ± 1.22 9.91 ± 1.79 8.34 ± 1.36*
Day 7 14.52 ± 1.81 13.95 ± 1.95 14.59 ± 2 08 11.76 ± 2.14*
Day 14 26.69 ± 4.22 25.56 ± 3.82 26.40 ± 3.86 21.90 ± 3.77*
Day 21 45.60 ± 4.22 42.86 ± 7.49 44.65 ± 7.14 35.59 ± 7.26*
Day 28 78.32 ± 9.91 73.79 ± 12.62 78.88 ± 11.37 65.86 ± 12.71*
females
Day 0/1 6.33 ± 0.42 6.18 ± 0.56 6.43 ± 0.63 5.94 ± 0.51*
Day 4 9.46 ± 0.91 9.17 ± 1.24 9.45 ± 1.40 8.03 ± 1.33*
Day 7 13.70 ± 1.58 13.17 ± 2.05 13.75 ± 2.36 11.38 ± 2.03*
Day 14 25.55 ± 4.26 24.25 ± 4.17 24.98 ± 4.26 21.07 ± 3.71*
Day 21 43.38 ± 6.07 41.8 ± 7.70 42.51 ± 6.96 34.07 ± 7.26*
Day 28 73.57 ± 9.16 68.99 ± 12.60 73.17 ± 11.38 61.85 ± 12.18*
* p< 0.05 (Dunnett-test)
Table 8 Selected relative organ weights of offspring (%, mean values and standard deviations)
Group Control 20 mg/kg 120 mg/kg 720 mg/kg
Heart  males 0.5396 ± 0.0754 0.5858 ± 0.0810* 0.5727 ± 0.0808 0.6135 ± 0.0750*
females 0.5357 ± 0.0688 0.5701 ± 0.0793 0.5770 ± 0.0930 0 5823 ± 0.0604*
Kidney males 1.419 ± 0.120 1.462 ± 0.158 1.421 ± 0.120 1.551 ± 0.141*
females 1.443 ± 0.102 1.468 ± 0.315 1.463 ± 0.103 1.536 ± 0.147*
Brain  males 1.9647 ± 0.2751 2.141 ± 0.3488* 2.0290 ± 0.3020 2.3799 ± 0.4206*
females 2.0406 ± 0.2921 2.1350 ± 0.3288 2.1193 ± 0.3733 2.3916 ± 0.4123*
Liver  males 5.967 ± 0.512 5.762 ± 0.494 5.621 ± 0.877 6.127 ± 0.398
females 6.092 ± 0.562 5.793 ± 0.550* 5.942 ± 0.470 6.149 ± 0.451
*p < 0.05 (Dunnett-test, two-tailed)

Applicant's summary and conclusion

Conclusions:
The NOAEL for general, systemic toxicity for males is 120 mg/kg/d, based on increased kidney and prostate weights and reduced body weights. The increase in liver weights is likely to be an adaptive response to the increased metabolic activity of the liver and can not be regarded as a toxic effect.

The NOAEL for general, systemic toxicity for parental females is also 120 mg/kg bw /d based on reduced body weight gain, reduced food consumption and increased organ weights (liver, and kidney) in the high dose group.

The NOAEL for reproductive toxicity is considered to be 720 mg/kg bw /d for the parental rats since no changes in reproductive parameters was observed.

The NOAEL for developmental toxicity in the F1 progeny is 720 mg/kg bw/d since effects on litter size, pup weight, pup survival and organ weights are secondary to maternal toxicity and reduced pup weight.
Executive summary:

The effects on fertility of the test material was evaluated in an OECD 415 one-generation study in rats. Animals received the test material via oral gavage in doses up to 720 mg/kg/d. Treatment related effects included organ weight effects (kidney & liver; males all dose groups and females high dose group only) and reduced food intake. Prostate weights were increased in high dose males as well.

No effects on reproductive parameters were observed. Mating performance and fertility were unaffected up to a dose level of 720 mg/kg. In the high dose group litter size, pup weight, pup survival and organ weights were affected, but these effects are secondary to maternal toxicity.