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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:
1991
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable well-documented study report which meets basic scientific principles.

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:
Male rats were given 0, 750, 1500 or 3000 mg/kg neat JP-8 daily by gavage for 70 days prior to mating with naive females to assess fertility and sperm parameters.
GLP compliance:
not specified
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
The JP-8 jet fuel was supplied by the U.S. Air Force (AFRL Propulsion Directorate, Wright- Patterson AFB, OH). The fuel met the requirements of Military Specification MIL-T-83133A. JP-8 was administered by gavage without a vehicle (neat). Control animals were dosed with 1.0 mL distilled water under the same conditions as test groups. Volumes to be administered each day were calculated from the individual daily body weights and the density of the test material (0.81 g/mL).

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Breeding Labs, Kingston, NY
- Weight at study initiation: (P) Males: 180 to 220 g
- Diet (e.g. ad libitum): Formula 5008, Ralston Purina, St. Louis, MO, ad libitum
- Water (e.g. ad libitum):ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-25
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on exposure:
JP-8 was administered by gavage without a vehicle (neat). Control animals were dosed with 1.0 mL distilled water under the same conditions as test groups. Volumes to be administered each day were calculated from the individual daily body weights and the density of the test material (0.81 g/mL).
Details on mating procedure:
In order to stagger delivery dates, male rats were paired with more than one female rat between 70 and 90 days of dosing with JP-8. Male rats were gavaged during cohabitation and returned to individual cages after successful mating. Exposure was continued until the rats were euthanized by carbon dioxide overdose at 90 days
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
90 days
Frequency of treatment:
daily; 7 days/ week
Details on study schedule:
The rats were given 0 (control), 750, 1500 or 3000 mg/kg JP-8 daily by gavage for 70 days prior to mating with naive females. Male rats were cohabitated with one female at a time. In order to stagger delivery dates, male rats were paired with more than one female rat between 70 and 90 days of dosing with JP-8. Male rats were gavaged during cohabitation and returned to individual cages after successful mating.
Doses / concentrations
Remarks:
Doses / Concentrations:
0 (control), 750, 1500 or 3000 mg/kg
Basis:
actual ingested
No. of animals per sex per dose:
20 males
Control animals:
yes, sham-exposed
Positive control:
none

Examinations

Parental animals: Observations and examinations:
body weight and mortality
Oestrous cyclicity (parental animals):
not examined
Sperm parameters (parental animals):
The epididymides were collected from each male rat at necropsy. The epididymides were then minced in phosphate buffered saline with bovine serum albumin; the resulting sperm suspension was videotaped in a Petroff Hauserr chamber. Determinations were made from the videotape using the CellSoft Automated Semen Analyzer (CRYO Resources, Ltd., New York, NY). Motility parameters measured by the CellSoft Analyzer were: sperm concentration, motile sperm concentration, percent motility, velocity, linearity, maximum amplitude of lateral head displacement (ALH), mean ALH and beat/cross frequency. The CellSoft Analyzer also measured the following parameters: mean radius, number of circular cells, percent circular cells/motile cells and percent circular cells/all cells.
Litter observations:
not examined
Postmortem examinations (parental animals):
not examined
Postmortem examinations (offspring):
not examined
Statistics:
Statistical Analyses for General Toxicity Data
Adult body weights, organ weights and urine metabolites were analyzed by an ANOVA with multiple comparisons. Clinical chemistry results, hematology values, urinalysis data and severity of pathological changes were compared using an ANOVA. The level of significance was accepted at p<0.05 unless stated otherwise.

Statistical Analyses for Reproductive Measures
A one-factor (dose) or two-factor (dose and pup sex) analysis of variance (ANOVA) was performed for continuous variables. Error terms used were either dam(dose) or pup sex and dam(dose). One-way ANOVA was used with gestation lengths, sperm parameters and litter sizes while two-way was used for pup weights. Post-hoc paired comparisons of dose used two-tailed t-tests with pooled error.

For categorical variables, a Chi-square test of proportions was used to determine differences among the doses. Chi-square tests were used for pregnancy rates and percent viability. Posthoc paired comparison for the viability parameter was performed with Fisher's Exact test. The level of significance was accepted at p<0.05 unless stated otherwise.
Reproductive indices:
Unexposed females mated with dosed males were allowed to give birth in order to determine gestation length. Successful mating (gestation day 0) was determined by presence of copulatory plug or sperm in a vaginal contents smear. Pregnancy rate (%) and gestation duration (days) were recorded for all dams. All rats were euthanized by carbon dioxide overdose.
Offspring viability indices:
not examined

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:
not examined
Histopathological findings: non-neoplastic:
not examined
Other effects:
not examined

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
not examined

Details on results (P0)

There were no clinical signs of toxicity other than changes in body weight. Body weights in male rats decreased in a dose dependent manner during 90 days gavage exposure to JP-8. A delay in starting dosing after randomly assigning rats to groups resulted in the 3000 mg/kg/day dose group being significantly heavier during the first weeks (p<0.05). Body weights were not different between groups from days 9 through 25 of exposure. From day 26 through the end of the study, the 3000 mg/kg/day dose group was significantly lighter than the control group (p<0.05). Mortality was limited to one male rat in the 750 mg/kg/day dose group.

Gestation parameters for unexposed females mated with treated males are shown below (Table 1). Pregnancy rates and gestation lengths were not adversely affected by paternal gavage exposure to JP-8. These parameters were not significantly different between dose groups. As a whole, these dams had low pregnancy rates, including the controls.

Epididymal sperm samples from males exposed by gavage to 0, 750, 1500 and 3000 mg/kg/day JP-8 for 90 days were evaluated using the CellSoft Automated Semen Analyzer. Table 2 contains sperm values for each dose group. The number of male rats per group ranged from 20 to 23. Outliers were removed after rigorous statistical analysis and were not related to dose. Significant differences were not found under any condition of analysis.

Effect levels (P0)

Dose descriptor:
NOAEL
Remarks:
fertility
Effect level:
>= 3 000 mg/kg bw/day (actual dose received)
Sex:
male

Results: F1 generation

General toxicity (F1)

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

Overall reproductive toxicity

Reproductive effects observed:
not specified

Any other information on results incl. tables

TABLE 1. GESTATION PARAMETERS OF UNEXPOSED DAMS MATED TO MALE RATS

Dose Group Number of Dams Pregnancy Rate Gestation Length
mg/kg/day (n) (%) mean (+/- SE)
0 36 47 21.24 (0.26)
750 38 39 21.07 (0.18)
1500 42 57 21.08 (0.15)
3000 32 53 21.41 (0.12)

TABLE 2. SPERM PARAMETERS (MEAN ± SE) IN MALE RATS EXPOSED

Parameter 0 mg/kg/day 750 mg/kg/day 1500 mg/kg/day 3000 mg/kg/day
  (n=21) (n=21) (n=23) (n=20)
Percent Motile 25.06 +/- 2.07 29.60 +/- 2.26 25.10 +/- 1.59 24.60 +/- 2.01
Conc. Motile (million/mL) 0.21 +/- 0.02 0.19+/- 0.02 0.20 +/- 0.02 0.16 +/- 0.02
Mean Velocity (um/s) 112.21 +/- 4.14 122.59 +/- 5.64 117.85 +/- 5.09 117.04 +/- 4.84
Mean Linearity 3.74 +/- 0.14 3.70 +/- 0.21 4.27 +/- 0.16 4.04 +/- 0.21
Max ALH (um) 4.04 +/- 0.19 4.28 +/- 0.24 4.06 +/- 0.15 4.00 +/- 0.19
Mean ALH (um) 3.44 +/- 0.15 3.64 +/- 0.18 3.47+ 0.1 3.41 +/- 0.14
Beat/Cross Frequency Hz (1/s) 10.42 +/- 0.28 10.34 +/- 0.23 10.86 +/- 0.19 10.70 +/- 0.27
Avg Radius (um) 16.04 +/- 1.19 15.67 +/- 0.92 15.93 +/- 1.22 13.79 +/- 0.93
Circular % of Motile 14.18 +/- 1.43 13.96 +/- 1.72 17.26 +/- 1.86 16.38 +/- 2.3
Circular % of All Cells 3.87 +/- 0.63 4.33 +/- 0.71 4.23 +/- 0.5 4.30 +/- 0.72

Applicant's summary and conclusion

Conclusions:
The NOAEL >=3000 mg/kg/day for male rat fertility.
Executive summary:

Male rats were given 0, 750, 1500 or 3000 mg/kg neat JP-8 daily by gavage for 70 days prior to mating with naive females to assess fertility and sperm parameters. Males were allowed to mate while continuing to receive treatment. Aside from a decrement in male body weight, no clinical signs were observed. There were no statistical differences noted in any reproductive parameter measured. The reproductive NOAEL >= 3000 mg/kg/day for male rats.