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Effects on fertility

Description of key information

Short description of key information:
No guideline studies were identified for effects on reproductive function.
Gonadal histopathology and/or sperm parameters (counts; morphology) were among endpoints included in a subchronic study with aerosol inhalation exposure to diesel fuel. No effects on reproductive organs were observed and the NOAEL was 1710 mg/m3 (for aerosolised diesel fuel).

Effect on fertility: via oral route
Endpoint conclusion:
no study available
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available (further information necessary)
Additional information

No guideline studies were located that have examined the potential impact on reproductive function. Some indication of the likely effect of a test substance on reproductive organs can be gained from the results of repeated-dose toxicity studies with closely related materials, as summarized in Table 1.

 

Table 1. Summaries of data on reproductive organs from subchronic studies with related substances. (Robust study summaries are provided in Section 7.5 Repeated Dose Toxicity)

Test Material

Route, Species, Doses, Exposure Regimen

Endpoints

Results

Reference

Diesel fuel

Inhalation. Rat.

350, 880, 1710 mg/m3

4 hr/day, twice/wk, 13 wk

Weight and histopathology of testes

No treatment-related effect noted

Lock et al, 1984

Cherry point diesel fuel No. 2

CAS 68476-34-6

Dermal. Rat.

0.5, 2.0, 5.0 ml/kg

5 d/wk, 4 wk

Weight and histopathology of testes and ovaries

No treatment-related effect noted

ARCO, 1986a

Naval distillate Watson

CAS 68334-30-5

Dermal. Rat.

0.25, 2.0, 5.0 ml/kg

5 d/wk, 4 wk

Weight and histopathology of testes and ovaries

No effect noted. Testicular degeneration in 2/10 males considered spontaneous.

ARCO, 1986b

Watson diesel fuel #2

68476-34-6

Dermal. Rat.

0.5, 2.0, 5.0 ml/kg

5 d/wk, 4 wk

Weight and histopathology of testes and ovaries

No treatment-related effect noted

ARCO, 1986c

Diesel fuel

CAS 68476-34-6

Dermal. Rat.

0.5, 1.0, 2.5, 5.0, 10.0 ml/kg

5 d/wk, 4 wk

Weight and histopathology of testes and ovaries

No treatment-related effect noted

ARCO, 1988

Naval distillate

CAS 68334-30-5

Dermal. Rat.

0.0001, 0.005, 0.5 ml/kg

5 d/wk, 4 wk

Weight and histopathology of testes and ovaries

No treatment-related effect noted

ARCO, 1992a

Naval distillate

CAS 68334-30-5

Dermal. Rat.

0.50 ml/kg

5 d/wk, 4 wk

Weight of testes and ovaries. No histopathology was done.

No treatment-related effect noted

ARCO, 1994a

Diesel fuel

(F-237)

68334-30-5

Dermal. Rat.

0.01, 0.10, 1.00 ml/kg,

5 d/wk, 13 wk

Weight of testes and ovaries. Histopathology of testis, ovary, prostate, uterus

No treatment-related effect noted

ARCO, 1994b

Based on test results from the above studies, it is considered unlikely that exposure will affect reproductive performance.

The cracked gas oils are regulated as Category 1B carcinogens (H350) according to EU CLP Regulation (EC No. 1272/2008), therefore further investigations of effects on fertility are not proposed.


Effects on developmental toxicity

Description of key information
Results of two inhalation developmental toxicity studies indicate a NOAEC >2,110 mg/m3. Maternal and foetal NOAELs of 125 mg/kg body weight/day were established from dermal prenatal developmental toxicity studies (equivalent or similar to OECD 414). In the dermal studies the only fetal effect was reduced pup weight, which occurred at dose levels that also caused maternal toxicity.  There are no acceptable developmental studies following oral exposure.
Link to relevant study records

Referenceopen allclose all

Endpoint:
developmental toxicity
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
1978-11-13, end date not specified
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: This study is classified as reliable without restrictions because it was well documented and performed similar to OECD 414. No information on GLP.
Justification for type of information:
See read-across justification provided in section 13.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
There were few details about the test compound.
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
other: CRL:COBS CD (SD)BR
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Breeding Laboratories, Wilmington, Massachusetts
- Age at study initiation: 11 weeks old
- Weight at study initiation: Not reported
- Housing: Individually except during mating
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum):ad libitum, acidified (pH 2.5) water
- Acclimation period: 13 days


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


IN-LIFE DATES: From:1978-11-13 To: Not reported
Route of administration:
inhalation: vapour
Type of inhalation exposure (if applicable):
whole body
Vehicle:
unchanged (no vehicle)
Details on exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Warm glass wool filled flask with compressed air
- Method of holding animals in test chamber: Stainless steel and plexiglass containers
- Source and rate of air: Compressed air at an unspecified rate
- Method of conditioning air: Not reported
- Temperature, humidity, pressure in air chamber: Under negative pressure at the following temperatures: control=26.7+/-1.04 degrees Celsius; 100 ppm=25.5+/-1.30 degrees Celsius; 400 ppm=26.8+/-1.00 degrees Celsius
- Air flow rate: 28.3 litres per minute
- Air change rate: Not reported
- Method of particle size determination: Not determined
- Treatment of exhaust air: Not reported

TEST ATMOSPHERE
- Brief description of analytical method used: Scott Model 216 Hydrocarbon Analyzer
- Samples taken from breathing zone: no
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Chamber concentrations were monitored hourly using a Scott Model 216 Hydrocarbon Analyzer calibrated with known concentrations of diesel fuel. Methane was used as an internal standard.
Details on mating procedure:
- Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: Not reported
- Length of cohabitation: Not reported
- Verification of same strain and source of both sexes: yes
- Proof of pregnancy: vaginal plug referred to as day 0 of pregnancy
Duration of treatment / exposure:
10 days (gestational days 6 through 15)
Frequency of treatment:
6 hours a day
Duration of test:
14 days
No. of animals per sex per dose:
Twenty pregnant dams per dose
Control animals:
yes, sham-exposed
Details on study design:
- Dose selection rationale: Stated as selected by API.
- Rationale for animal assignment (if not random): Assigned sequentially to the dose groups

Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily
- Cage side observations included general appearance, behaviour and condition.


DETAILED CLINICAL OBSERVATIONS: No


BODY WEIGHT: Yes
- Time schedule for examinations: On gestational days 0, 6, 15, and 20


FOOD CONSUMPTION: Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No



WATER CONSUMPTION: No


POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day # 20
- Organs examined: Visceral and thoracic organs
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No
- Number of corpora lutea: No
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
Fetal examinations:
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: one-third per litter
- Skeletal examinations: Yes: two-thirds per litter
- Head examinations: Yes: one-third per litter
Statistics:
Continuous parameters were analyzed with Dunnet's t-test. A 2x2 contingency table with Yates correction was used for ratio data. Discontinuous data (e.g., number of abnormal foetuses per litter) were analyzed using Wilcoxon Rank Sum test.
Indices:
Nidation index, liver and dead foetuses, implantations, pup weight, sex ratio, and abnormalities
Historical control data:
Historical data on 54 litters was provided for reproductive performance. The concurrent control was considered to be similar to the historical control.
Details on maternal toxic effects:
Maternal toxic effects:no effects

Details on maternal toxic effects:
There were no deaths during the course of the study and all females were normal in appearance. Mean body weights indicated no significant differences between control and treated pregnant rats, although the food consumption of the high-dose group was significantly lower (p < 0.05) than the controls during treatment. Examination at necropsy revealed two high-dose females with dark, mottled lungs. One of these animals had a rough spleen but no further description was provided for this and the authors did not consider either of these effects to be treatment related.

Key result
Dose descriptor:
NOAEC
Effect level:
401.5 ppm (analytical)
Based on:
other:
Remarks:
overall effects
Basis for effect level:
other: maternal toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
Examination of the offspring revealed no visible abnormalities except for the occurrence of subcutaneous haematomas in 1/188, 4/227 and 4/188 in the control, low-, and high-dose groups, respectively. All litters appeared normal and the sex ratio did not differ significantly between treated and control groups. Some unusual skeletal variations related to retarded bone ossification were observed, including reduced ossification of the parietal bones and the frontal bones, and wavy ribs. These changes were noted in 7 and 17 pups from 3 and 8 litters of the control and low-dose animals only, and were not dose related. The authors concluded that neither the frequency nor the character of these changes indicated an adverse effect on foetal growth and development or a teratogenic potential. Other skeletal variations were considered to be common to animals of this strain.
Key result
Dose descriptor:
NOAEC
Effect level:
401.5 ppm (analytical)
Based on:
other:
Remarks:
overall effects
Basis for effect level:
other: developmental toxicity
Abnormalities:
not specified
Developmental effects observed:
not specified
Conclusions:
Exposure of pregnant rats to diesel fuel (vapour) did not result in compound-induced maternal toxicity, teratology, variation in sex ratio, embryo toxicity or inhibition of foetal growth and development. Therefore the NOAEC was 2,110 mg/m3, the highest concentration tested.
Executive summary:

The teratology of diesel fuel was examined by whole body exposure of pregnant female (CRL: COBS CD (SD) BR) rats to graded measured airborne (vapour) of 0, 101.8, or 401.5 ppm (equivalent to 0, 530, and 2110 mg/m3, respectively) on days 6 through to 15 of gestation. The pregnant female rats were approximately 11 weeks of age at the time of the first dose and were assigned sequentially to treatment groups of 20 animals each. They were exposed to diesel fuel for 6 hours/day in 0.25 m3 stainless steel and plexiglass chambers operated under negative pressure. The animals were assessed daily for body weight changes, food consumption, and clinical observations.

There were no deaths during the course of the study and all females were normal in appearance. Mean body weights indicated no significant differences between control and treated pregnant rats, although the food consumption of the high-dose group was significantly lower (p < 0.05) than the controls during treatment.

On day 20 of gestation, the study was terminated and the rats were anaesthetised with chloroform. Following anaesthesia, gross examination of the organs, examination of the uterus (number of implantation sites, live/dead foetuses, number of resorption sites) and foetal examinations (soft tissue and skeletal) were performed.

Examination at necropsy revealed two high-dose females with dark, mottled lungs. One of these animals had a rough spleen but no further description was provided for this and the authors did not consider either of these effects to be treatment related.

Diesel fuel did not appear to have any effect on the uterus and examination of the offspring revealed no visible abnormalities except for the occurrence of subcutaneous haematomas in 1/188, 4/227 and 4/188 in the control, low-, and high-dose groups, respectively. All litters appeared normal and the sex ratio did not differ significantly between treated and control groups. Some unusual skeletal variations related to retarded bone ossification were observed, including reduced ossification of the parietal bones and the frontal bones, and wavy ribs. These changes were noted in 7 and 17 pups from 3 and 8 litters of the control and low-dose animals only, and were not dose related. The authors concluded that neither the frequency nor the character of these changes indicated an adverse effect on foetal growth and development or a teratogenic potential. Other skeletal variations were considered to be common to animals of this strain.

 

The authors concluded that exposure of ratsto diesel fuel (vapour) did not result in compound-induced terata, variation in sex ratio, embryo toxicity or inhibition of foetal growth and development. Therefore the NOAEC was 2,110 mg/m3 (401.5 ppm), the highest concentration tested.

This study received a Klimisch score of 2 and is classified as reliable with restrictions because the study is well-documented and generally followed OECD 414 guidelines. No information was available on GLP.

Endpoint:
developmental toxicity
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: This study is classified as reliable with restrictions because the high-dose had to be discontinued due to maternal discomfort and was not available to determine developmental toxicity, but the study was well conducted and was GLP compliant.
Justification for type of information:
See read-across justification provided in section 13.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
No skeletal or visceral exams were conducted.
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Route of administration:
dermal
Vehicle:
acetone
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
20 days
Frequency of treatment:
daily
Duration of test:
20 days
No. of animals per sex per dose:
between 14 and 19 females
Control animals:
yes, concurrent vehicle
Details on maternal toxic effects:
Maternal toxic effects:yes

Details on maternal toxic effects:
dermal irritation, reduced body weight gain
Dose descriptor:
LOAEL
Effect level:
125 mg/kg bw/day
Basis for effect level:
other: maternal toxicity
Dose descriptor:
LOAEL
Effect level:
125 mg/kg bw/day
Basis for effect level:
other: developmental toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:yes

Details on embryotoxic / teratogenic effects:
reduced pup weight post parturition until day 4
Abnormalities:
not specified
Developmental effects observed:
not specified
Conclusions:
The maternal LOAEL is 125 mg/kg/day, based on dermal irritation. There is no maternal NOAEL. The developmental LOAEL is 125 mg/kg/day, based on reduced pup weight on lactational day 0. There is no developmental NOAEL. 
Executive summary:

In a developmental toxicity study, VDF diesel in acetone was dermally administered to 14 to 19 presumed pregnant rats/dose at dose levels of 0, 125, or 250 mg/kg bw/day from days 0 through 20 of gestation. Another group of 14 received 1000 mg/kg/day from days 0 to 5 of gestation.

 

Dermal irritation occurred in all treatment groups. Body weight, body weight gain, and food consumption was reduced in the 250- and 1000 -mg/kg/day groups. The maternal LOAEL is 125 mg/kg/day, based on dermal irritation. There is no maternal NOAEL. Pup weight was decreased on lactational day 0 in all dose groups and on lactational day 4 in the 250 -and 1000 -mg/kg/day groups. The developmental LOAEL is 125 mg/kg/day, based on reduced pup weight. There is no developmental NOAEL.

 

This study received a Klimisch score of two and is classified as reliable with restrictions because the high-dose had to be discontinued due to maternal discomfort and was not available to determine developmental toxicity, but the study was well conducted and was GLP compliant.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEC
2 110 mg/m³
Study duration:
subacute
Species:
rat
Quality of whole database:
One of two inhalation studies investigating the developmental toxicity of structurally related materials
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LOAEL
125 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
One of three dermal studies investigating the developmental toxicity potential of structurally related materials.
Additional information

Results are available from studies involving inhalation exposure of pregnant rats to either diesel oil or No.2 Home Heating Oil (a mixture of straight run gas oil and cracked gas oil) vapour. No treatment related effects on maternal health or foetal development were observed in either study, giving a NOAEC >2,110 mg/m3. The studies were well conducted and reported but the exposure period (GD 6-15) was less intensive than that recommended by current guidelines (GD 5-20) and only covered the period of organogenesis in the rat. The results do however indicate that inhalation exposure to gas oil vapour is unlikely to adversely impact foetal development.

In the first inhalation teratology study (API 1979), diesel fuel was administered via whole body exposure of pregnant female (CRL: COBS CD (SD) BR) rats to graded measured airborne (vapour) of 0, 101.8, or 401.5 ppm (equivalent to 0, 530, and 2110 mg/m3, respectively) on days 6 through to 15 of gestation. The pregnant female rats were approximately 11 weeks of age at the time of the first dose and were assigned sequentially to treatment groups of 20 animals each. There were no deaths during the course of the study and all females were normal in appearance. Mean body weights indicated no significant differences between control and treated pregnant rats, although the food consumption of the high-dose group was significantly lower (p < 0.05) than the controls during treatment. Diesel fuel did not appear to have any effect on the uterus and examination of the offspring revealed no visible abnormalities except for the occurrence of subcutaneous haematomas in 1/188, 4/227 and 4/188 in the control, low-dose, and high-dose groups, respectively. All litters appeared normal and the sex ratio did not differ significantly between treated and control groups. The conclusion of the authors was that exposure of rats to diesel fuel (vapour) did not result in compound-induced abnormalities, variation in sex ratio, embryo toxicity or inhibition of foetal growth and development. Therefore the NOAEC is >2,110 mg/m3 (401.5 ppm), the highest concentration tested.

In the second study, the effect of inhalation of No.2 Home Heating Oil, a mixture of straight run gas oil and cracked gas oil, ( API 1979) vapour on foetal development was tested in three groups of pregnant female CRL; COBS CD (SD) BR rats. The 11 week old females were randomly assigned into groups of 20 animals each and treated (whole body) with measured concentrations of 0 (control group), 86.9 or 408.8 ppm (equivalent to 460 or 2,150 mg/m3) test substance from days 6 to 15 of gestation for 6 hours each day. There were no treatment related deaths or changes in appearance of dams throughout the duration of the study. One female delivered before the computed gestation day 20 and her pups were examined but not included in statistical analysis. There were no statistically significant changes in uterine contents. Subcutaneous haematomas were observed in all groups – 7, 4 and 3 in control, low-dose and high-dose groups respectively. There were no statistically significant or treatment related adverse effects on reproductive or developmental toxicity in this study leading to a NOAEC of > 2,150 mg/m3, the highest dose tested.

In a developmental toxicity screening study, straight run diesel in acetone was dermally administered to 14 or 15 presumed pregnant rats/dose at dose levels of 0, 125, or 250 mg/kg bw/day from days 0 through 20 of gestation. Another group of 15 rats received 1000 mg/kg/day from days 0 to 5 of gestation ( ARCO 1994). Dermal irritation occurred in all treatment groups. Body weight, body weight gain, and food consumption was reduced in the 250- and 1000 -mg/kg/day groups. The maternal LOAEL is 125 mg/kg/day, based on dermal irritation.  There is no maternal NOAEL. Pup weight was decreased on lactation days 0 and 4 in the 250 -mg/kg/day group. There were no developmental effects observed in the 1000 -mg/kg/day for 6 days. The developmental LOAEL is 250 mg/kg/day, based on reduced pup weight. The developmental NOAEL is 125 mg/kg/day.  

In an additional supporting developmental toxicity screening study, VDF diesel in acetone was dermally administered to 14 or 19 presumed pregnant rats/dose at dose levels of 0, 125, or 250 mg/kg bw/day from days 0 through 20 of gestation. Another group of 14 received 1000 mg/kg/day from days 0 to 5 of gestation (ARCO 1994). Dermal irritation occurred in all treatment groups. Body weight, body weight gain, and food consumption was reduced in the 250- and 1000 -mg/kg/day groups. The maternal LOAEL is 125 mg/kg/day, based on dermal irritation. There is no maternal NOAEL. Pup weight was decreased on lactation day 0 in all dose groups and on lactation day 4 in the 250 -and 1000 -mg/kg/day groups. The developmental LOAEL is 125 mg/kg/day, based on reduced pup weight. There is no developmental NOAEL.

These studies were classified as reliable with restriction, because the high-doses had to be discontinued due to maternal discomfort and were not available to determine developmental toxicity. The studies were well conducted and were GLP compliant.

In a further developmental embryo-fetal toxicity and teratogenic potential study, straight run diesel in acetone was dermally administered to 25 presumed pregnant rats/dose at dose levels of 0, 50, 150, or 300 mg/kg bw/day from days 0 through 19 of gestation (ARCO 1993). Skin reactions (erythema, oedema, atonia, and desquamation) occurred in all treatment groups. In the mid- and high-dose group, vocalization occurred in 5 or 6 of the animals. There was a slight decrease in body weight gain in the 50 mg/kg/day group with significant reductions occurring in the 150- and 300-mg/kg/day groups. Feed consumption was only reduced in the high-dose group. There were no treatment-related effects on reproduction or development (number of corpora lutea, implantations, litter size, live fetuses, resorptions, fetal body weight, or sex ratio). There were no gross external, skeletal, or visceral abnormalities observed. The maternal LOAEL is 50 mg/kg/day, based on dermal effects and reduced body weight gain. There is no maternal NOAEL. The developmental NOAEL is>300 mg/kg/day, based on no effects at the highest dose tested.

Studies are available to investigate the developmental toxicity of diesel fuel/heating oil following dermal and inhalation exposure. Although effects were observed on progeny weight in some dermal studies, the effects only occurred at dose levels that also caused maternal toxicity. Based on this information alone it is not considered possible to determine if the effects were direct on the foetus or caused indirectly by effects on the pregnant dam.

 

Justification for classification or non-classification

The cracked gas oils are regulated as Category 1B carcinogens (H350) according to EU CLP Regulation (EC No. 1272/2008), therefore further investigations of effects on fertility are not proposed.

Developmental studies only observed developmental effects (reduced pup weight) at doses that also caused maternal toxicity and it was not possible to determine whether the effects were direct on the foetus or caused indirectly by effects on the pregnant dam.

Classification for developmental toxicity is therefore not considered appropriate.