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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
fertility, other
Remarks:
based on test type (migrated information)
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
Not reported
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study is classified as reliable without restrictions because it is an acceptable and well-documented study report.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2000
Report Date:
2000

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 415 [One-Generation Reproduction Toxicity Study (before 9 October 2017)]
Principles of method if other than guideline:
This was a reproductive study performed in two parts. In the first part, males were treated for 70 to 90 days with 0 (1mL of distilled water), 750, 1500, or 3000 mg/kg/day of undiluted JP-8 jet fuel, then mated to untreated females (one female at a time). In the second part of the study, female rats were administered the test compound at doses of 0 (1mL of distilled water), 375, 750, or 1500 mg/kg/day undiluted JP-8 jet fuel for 90 -day prior to mating, through mating, gestation, delivery, and lactation for a total of 21 weeks.
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
other: low viscosity liquid hydrocarbon
Details on test material:
- Name of test material (as cited in study report): JP-8 jet fuel
- Other: The fuel met the requirements of Military Specification MIL-T-83133A.

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
- Source: Charles River Breeding Labs, Kingston, New York
- Weight at study initiation: 180 to 220 grams
- Housing: Individually, except during cohabitation (1 male with 1 female)
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 2 weeks


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 to 25 °C
- Humidity (%): Not reported
- Air changes (per hr): Not reported
- Photoperiod (hrs dark / hrs light): 12 hours dark/12 hours light

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on mating procedure:
- M/F ratio per cage: 1 to 1
- Length of cohabitation: Not reported
- Proof of pregnancy: vaginal plug or sperm in vaginal smear referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged (how): Not reported
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
Not applicable
Duration of treatment / exposure:
Males were treated for 70 to 90 days. Females were treated for 21 weeks.
Frequency of treatment:
Daily
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
Males: 750, 1500, or 3000 mg/kg/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
Females: 325, 750, or 1500 mg/kg/day
Basis:
actual ingested
No. of animals per sex per dose:
A minimum of 20 male rats per dose were used to test male fertility and a minimum of 35 female rats were used to test effects on female fertility.
Control animals:
yes, sham-exposed
Details on study design:
- Dose selection rationale: Doses were based on a previous 90-day study.
Positive control:
None reported

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Not reported
- Cage side observations examined were not reported.


DETAILED CLINICAL OBSERVATIONS: No data


BODY WEIGHT: Yes
- Time schedule for examinations:Daily


FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No


OTHER: Hematology (table 1), clinical chemistry (table 2), and urinalysis (table 3) was performed on a maximum of 10 females per treatment. In the same rats used for haematology and clinical chemistry, the brain, kidneys, liver, spleen, and ovaries were weighed.
Oestrous cyclicity (parental animals):
Not performed
Sperm parameters (parental animals):
Parameters examined in P male parental generations:
sperm count in epididymides, sperm motility, velocity, linearity, maximum and mean amplitude of lateral head displacement, beat/cross frequency, mean radius, number of circular cells, % circular cells/motile cells, % circular cells/all cells
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 5 postpartum: Four male and four female pups

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, stillbirths, live births, weight gain

GROSS EXAMINATION OF DEAD PUPS:no
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals were sacrificed on day 90 of treatment (mating occurred during days 70 to 90 of treatment.
- Maternal animals: All surviving animals were sacrificed 1 day after weaning.

GROSS NECROPSY
- Gross necropsy details were not reported.

HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table 4 were prepared for microscopic examination and weighed, respectively.
Postmortem examinations (offspring):
No postmortem examinations were conducted on the offspring.
Statistics:
General toxicity data: an ANOVA with or without multiple comparisons
Reproductive measures: a one- or two-factor ANOVA and a post-hoc comparison of dose using two-tailed t-test with pooled error was used for continuous data; a Chi-square test of proportions followed by a Fischer's Exact test was used for categorical data
Reproductive indices:
Pregnancy rate, gestation duration, litter size
Offspring viability indices:
Live and dead offspring, pup weight on postnatal days 1, 4, 7, 14, 21, and 90

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
no effects observed
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
Histopathological findings: non-neoplastic:
effects observed, treatment-related

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
no effects observed
Reproductive performance:
no effects observed

Details on results (P0)

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS): There were no changes in clinical signs or mortality.


BODY WEIGHT AND WEIGHT GAIN (PARENTAL ANIMALS): Body weights in male rats were decreased in a dose-dependent manner. Terminal body weights were approximately 545 grams, 520 grams, 475 grams, and 315 grams in the control, 750, 1500, and 3000 mg/kg/day, respectively (results were approximated from a figure). In females, body weight was only significantly reduced in the high-dose group, but the differences were not significant at terminal sacrifice. The body weight in females at 20 weeks (1 week before sacrifice) was approximately 400 grams, 385 grams, 382 grams, and 335 grams in the control, 375, 750, and 1500 mg/kg/day, respectively.


HAEMATOLOGY (PARENTAL ANIMALS): Haematology was not measured in the males and no effects were noted in the females.


CLINICAL CHEMISTRY (PARENTAL ANIMALS): Clinical chemistry was not measured in the males and no effects were noted in the females.


URINALYSIS (PARENTAL ANIMALS): Urinalysis was not measured in the males and no effects were noted in the females.


ORGAN WEIGHTS (PARENTAL ANIMALS): Absolute and relative liver weights were increased in mid- and high-dose females (Table 5), but were not accompanied by any histological findings.


HISTOPATHOLOGY (PARENTAL ANIMALS): The test compound caused perianal dermatitis (high-dose only) and stomach hyperplasia (mid- and high-dose) in the female rats (Table 6).

OTHER FINDINGS (PARENTAL ANIMALS): There were no treatment-related effects on reproduction or sperm parameters in males. There were no effects on reproduction, gestation, or litter size in females.

Effect levels (P0)

open allclose all
Dose descriptor:
NOAEL
Effect level:
750 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: body weight
Dose descriptor:
NOAEL
Remarks:
reproduction
Effect level:
>= 3 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: duration of pregnancy; pregnancy rate; sperm characterization
Dose descriptor:
NOAEL
Remarks:
reproduction
Effect level:
>= 1 500 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: duration of pregnancy; live birth index; pregnancy rate; litter size; litter weight

Results: F1 generation

General toxicity (F1)

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

Details on results (F1)

VIABILITY (OFFSPRING): There were no effects on offspring viability.

BODY WEIGHT (OFFSPRING): There was a dose-related decrease in pup weight that was significant in the 750 mg/kg/day group on postnatal day 4 only and in the 1500 mg/kg/day group from postnatal day 4 through postnatal day 21 but had recovered by postnatal day 90.

Effect levels (F1)

Dose descriptor:
NOAEL
Generation:
F1
Effect level:
750 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: pup weight

Overall reproductive toxicity

Reproductive effects observed:
not specified

Any other information on results incl. tables

The decreased body weight in males was probably related to nephropathy that is typically produced in male rats exposed to hydrocarbon fuels. Although there were changes in liver weight in the females, this was not accompanied by any changes in clinical chemistry or liver histopathology so is likely not of biological significance.
Table 5
 
Body weight and liver weights in female rats
  Control 325 mg/kg/day 750 mg/kg/day 1500 mg/kg/day
  n=10 n=7 n=8 n=8
Terminal Body weight 351.0 ± 12.6 354.3 ± 9.0 357.5 ± 6.5 348.8 ± 16.3
Absolute liver weight 14.8 ± 0.8 15.7 ± 0.6 17.8 ± 0.7 * 19.1 ± 0.7 **
Liver to body weight 4.2 ± 0.1 4.4 ± 0.1 5.0 ± 0.2 ** 5.5 ± 0.1 **
Liver to brain weight 754.9 ±38.9 823.6 ± 39.8 903.1 ± 31.8 * 993.6 ± 46.3 **
* p<0.05
** p<0.01
Table 6
Incidence and severity significant histopathological findings in female rats
  Control 325 mg/kg/day 750 mg/kg/day 1500 mg/kg/day
  n=8 n=4 n=5 n=6
Perianal dermatitis 0 0 0 83% (1.2) *
Anal hyperplasia 0 0 0 50% (0.7)
Stomach gastritis 0 50% (0.5) 20% (0.2) 0
Stomach hyperplasia 0 75% (0.8) 100% (1.6) * 100% (1.7) *
* p<0.05

Applicant's summary and conclusion

Conclusions:
The study LOAEL for systemic effects is 1500 mg/kg/day and the NOAEL for systemic effects is 750 mg/kg/day, based on reduced body weight in dams and in pups. The LOAEL for adult males rats exposed to JP-8 orally was 750 mg/kg/day due to changes in clinical pathology, body weight, organ weights and the same irritation seen in female rats. The reproduction NOAEL was 3000 and 1500 mg/kg/day in males and females, respectively.
Executive summary:

This was a reproductive study performed in two parts. In the first part, males were treated for 70 to 90 days with 0 (1mL of distilled water), 750, 1500, or 3000 mg/kg/day of undiluted JP-8 jet fuel, then mated to untreated females (one female at a time). Males were gavaged throughout the cohabitation period and were returned to their individual cage after successful mating. In the second part of the study, female rats were administered the test compound at doses of 0 (1mL of distilled water), 375, 750, or 1500 mg/kg/day undiluted JP-8 jet fuel for 90 -day prior to mating, through mating, gestation, delivery, and lactation for a total of 21 week. During mating, they were housed with untreated males. Litters were standardized to 4 male pups and 4 female pups on postnatal day 5. Although those pups were used for neurobehavioral tests, the data was not provided in the study report.

There were no effects on clinical signs or mortality in either sex. Haematology, clinical chemistry, and urinalysis were measured only in females without any effects noted. Body weights in male rats were decreased in a dose-dependent manner and was likely related to nephropathy that is specific in male rats treated with hydrocarbons. In females, body weight was only significantly reduced in the high-dose group. Absolute and relative liver weights were increased in mid- and high-dose females, but were not likely biologically significant due to the lack of changes in clinical chemistry or histopathology in the liver. The test compound caused perianal dermatitis (high-dose only) and stomach hyperplasia (mid- and high-dose) in the female rats (histopathology was not conducted on the males for this study because previous studies have indicated nephropathy). There were no treatment-related effects on reproduction or sperm parameters in males. There were no effects on reproduction, gestation, or litter size in females. There was a dose-related decrease in pup weight that was significant in the 750 mg/kg/day group on postnatal day 4 only and in the 1500 mg/kg/day group from postnatal day 4 through postnatal day 21 but had recovered by postnatal day 90.

The study LOAEL for systemic effects is 1500 mg/kg/day and the NOAEL for systemic effects is 750 mg/kg/day, based on reduced body weight in dams and in pups. The LOAEL for adult males rats exposed to JP-8 orally was 750 mg/kg/day due to changes in clinical pathology, body weight, organ weights and the same irritation seen in female rats.Changes in male rats may be complicated by the male rat-specific nephropathy produced after exposure to hydrocarbon fuels such as JP-4 and JP-8. The reproduction NOAEL was 3000 and 1500 mg/kg/day in males and females, respectively.

This study received a Klimisch score of 1 and is classified as reliable without restrictions because it is an acceptable and well-documented study report.