Extract obtained from defatted powder of Theobroma cacao (Malvaceae) by extraction with water and ethanol

Toxicity to reproduction

Administrative data

Endpoint:

fertility, other

Remarks:

based on test type (migrated information)

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: A published study containing sufficient details to regard it as reliable for use in hazard assessment.Limited experimental detail provided.

Data source

Reference

Reference Type:

publication

Title:

Rat Offspring Sired by Males Treated with Alcohol.

Author:

Abel, E.L.

Year:

1993

Bibliographic source:

Alcohol 10(3) :237-242

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 415 [One-Generation Reproduction Toxicity Study (before 9 October 2017)]

Deviations:

yes

Remarks:

untreated females, rationale for doses selected not provided

Principles of method if other than guideline:

Method: other

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Automatically generated during migration to IUCLID 6, no data available

IUPAC Name:

Automatically generated during migration to IUCLID 6, no data available

Details on test material:

no further data.

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding laboratory
- Age at study initiation: 2 Months old at acquisition
- Diet: ad libitum.
- Water: ad libitum.
- Acclimation period: 2 weeks.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 +/- 1 degree C
- Humidity: 45 +/- 5%
- Photoperiod: 12 hr:12 hr light:dark cycle

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: distilled water only

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

Exposure period: 9 weeks
Premating exposure period (males): 9 weeks
Food and water was removed at the time of first intubation at 9am and returned at 5pm.

Frequency of treatment:

twice per day

Details on study schedule:

After treatment, each male was paired with 2 untreated 75-90 day old females. Mating was confirmed by presence of plugs. Females were allowed to deliver their litter.

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 2, 3 g/kg twice daily, total dose 0, 4, 6g/kg/day
Basis:
other: nominal dose

No. of animals per sex per dose:

groups of 20.

Control animals:

yes, concurrent no treatment

yes, concurrent vehicle

Details on study design:

Groups receiving 2 g/kg or 0 g/kg were paired fed with those receiving 3 g/kg and a fourth group received no gavage treatment.

Examinations

Litter observations:

STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: Yes
- If yes, maximum of 10 pups per female.

PARAMETERS EXAMINED
The following parameters were examined in offspring: Culled pups were subjected to brain and adrenal gland weight measurements. Offspring were weighed at 7, 14 and 21 days. At 7 days old, three males and 3 females from each of 6 litters were culled and their brain and adrenal gland weights determined. At 21 days, this was repeated with inclusion of more organ weights. Blood alcohol levels were determined after breeding was determined at 1,2,3 & 5 hours after dosing.

Statistics:

ANOVA and Duncan's Multiple Range tests on parametric data; Chi-square on non-parametric data.

Results and discussion

Results: P0 (first parental generation)

Details on results (P0)

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
There were no adverse effects on male reproductive performance and female fecundity was not affected.

ORGAN WEIGHTS (PARENTAL ANIMALS)

Effect levels (P0)

Dose descriptor:

NOAEL

Effect level:

> 6 000 mg/kg bw/day (actual dose received)

Sex:

male/female

Results: F1 generation

Details on results (F1)

VIABILITY (OFFSPRING)
Litter sizes and birth weights were not affected. Litter sizes and birth weights were not affected by paternal ethanol intake at either dose.

BODY WEIGHT (OFFSPRING)
There was a significantly higher number of female runts (bodyweight <5.5 g) in the groups sired by rats exposed to ethanol. There was also a significantly higher number of male runts in the groups sired by rats exposed to ethanol. In both cases, these were only statistically significant in comparison to the vehicle treated controls. There was a slight difference between the untreated control animals as the high dose group but this was not statistically significant. In fact the differences between the two controls reached statistical significance. Ethanol treatment in fathers had no effect on offspring growth rate.

ORGAN WEIGHTS (OFFSPRING)
Ethanol treatment at both levels resulted in a significant increase in absolute and relative adrenal gland weight but not in brain weight in comparison to the vehicle treated controls but not the untreated ones. Organ weights (both absolute and bodyweight relative) were unaffected at 7 days but significant reductions of absolute spleen and heart weight and relative spleen weight were noted at 21 days at the 3g/kg dose level (NOEL=2g/kg) and reported to be statistically significant. It should be noted again that this statistical difference is not apparent in comparison to the untreated controls. Also, the reductions in weight in the mid dose group as reported are even larger, but not reported to be statistically significant.

OTHER FINDINGS (OFFSPRING)
The % of males in litters sired by ethanol treated rats was significantly lower than the % sired by vehicle-treated fathers (p<0.04) although the difference with the non-intubated control was not significant.

Effect levels (F1)

1

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

4 000 mg/kg bw/day (actual dose received)

Sex:

male/female

Basis for effect level:

other: Reduction of spleen and heart weight observed at 21 days in comparison to concurrent vehicle control where effects reported to be statistically significant.

2

Dose descriptor:

NOEL

Generation:

F1

Effect level:

> 6 000 mg/kg bw/day (actual dose received)

Sex:

male/female

Basis for effect level:

other: in comparison to non treatment control

Overall reproductive toxicity

Reproductive effects observed:

not specified

Any other information on results incl. tables

The percentage of males in treatment groups was reported to be significantly lower than controls but this was thought by the authors to be a chance finding.

Applicant's summary and conclusion

Conclusions:

There was no effect on fertility in a group of 20 male rats given 3 g or 2 g/kg ethanol by oral intubation daily for nine weeks, achieving BAL's of 338±15 and 132±5mg/dL, respectively. Other conclusions remain uncertain.

Executive summary:

Male rats were treated with 2 or 3g/kg ethanol twice a day by oral intubation for 9 weeks. At the end of the treatment they were paired with two untreated females each. Pregnant females were allowed to deliver their litter. The brain and adrenal weights were measured in the culled animals at birth, on postnatal day 7, and finally on postnatal day 21 with additional organ weight measurements. Paternal alcohol exposure did not influence litter size, average birth weight per pup or postnatal bodyweights in offspring. However, it induced a significant increase in the number of runts in the highest dose group which suggests an influence of ethanol on individual sperm rather than on entire sperm production. Treatment with ethanol also induced an increase in spleen weight at birth that did not persist, and a decrease in heart and spleen weight at 21 days at the highest dose treated. However, it should be noted that no findings were statistically significantly different from both controls used and that for those end points where the treatment groups caused statistically significant findings, the two control groups also varied significantly. On this basis, the positive findings reported here should be given low weighting.