Registration Dossier
Registration Dossier
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EC number: 229-352-5 | CAS number: 6485-40-1
Endpoint summary
Administrative data
Key value for chemical safety assessment
Effects on fertility
Description of key information
Toxicity to Reproduction (Screening study):
In accordance with Column 2 of ANNEX VIII of the REACH regulation,
screening for reproductive/developmental toxicity study does not need to
be conducted as a pre-natal developmental toxicity study is available.
Toxicity to Reproduction (Two generation study/EOGRTS):
In accordance with Column 2 of ANNEX IX of the REACH regulation, a
two generation reproductive toxicity study or EOGRTS does not need not
to be conducted as the existing 90 day study does not indicate adverse
effects on reproductive organs or tissues.
Link to relevant study records
- Endpoint:
- screening for reproductive / developmental toxicity
- Data waiving:
- other justification
- Justification for data waiving:
- the study does not need to be conducted because a pre-natal developmental toxicity study is available
- Reproductive effects observed:
- not specified
- Endpoint:
- extended one-generation reproductive toxicity - basic test design (Cohorts 1A, and 1B without extension)
- Data waiving:
- study scientifically not necessary / other information available
- Justification for data waiving:
- other:
- Reproductive effects observed:
- not specified
Referenceopen allclose all
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
Additional information
Toxicity to Reproduction (Screening study):
In accordance with Column 2 of ANNEX VIII of the REACH regulation,
screening for reproductive/developmental toxicity study does not need to
be conducted as a pre-natal developmental toxicity study is available.
Toxicity to Reproduction (Two generation study?EOGRTS):
In accordance with Column 2 of ANNEX IX of the REACH regulation, a
two generation reproductive toxicity study or EOGRTS does not need not
to be conducted as the existing 90 day study does not indicate adverse
effects on reproductive organs or tissues.
Effects on developmental toxicity
Description of key information
Pre-natal developmental toxicity study (rat, oral): NOAEL (developmental): <125 mg/kg bw/day; LOAEL (developmental): 125 mg/kg bw/day;
NOAEL (maternal): 250 mg/kg bw/day (OECD 414/GLP)
Link to relevant study records
- Endpoint:
- developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 414 (Prenatal Developmental Toxicity Study)
- GLP compliance:
- yes
- Limit test:
- no
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Wanxiang International; 2017072401
- Expiration date of the lot/batch: 2020/07/24
- Purity test date: 991.6%; 2017/07/24
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Store in cool place. Keep container tightly closed in a dry and well-ventilated place. - Species:
- rat
- Strain:
- other: Wistar CRL
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River SPF breeding, supplied via VELAZ s.r.o., Czech Republic, RČH CZ 11760500
- Age at study initiation: 10 - 11 weeks
- Weight at study initiation:
- Housing: Animals were housed in plastic cages containing sterilized clean shavings of soft wood or sterilized LIGNOCEL (raw material - spruce; producer: J.Rettenmaier & Söhne, Germany).
- Diet: Complete pelleted diet for rats and mice in SPF breeding (Altromin Spezialfutter) was used (Altromin Spezialfutter GmbH & Co. KG, Germany) ad libitum.
- Water: Free access to drinking water
- Acclimation period: 12 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3°C.
- Humidity (%): 30-70 %
- Air changes (per hr): 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12-hour light/12-hour dark cycle - Route of administration:
- oral: gavage
- Vehicle:
- corn oil
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS: The test item was weighted into glass beaker and then a smaller volume of olive oil was added. This suspension was well mixed and then added with olive oil to the required volume. After this the application form was dissolving in ultrasonic bath for 10 minutes and then application form was mixed by stirred by magnetic stirrer for 30 minutes at 500 rpm before application and then during administration. The procedure for application form preparation was taken from the analytical report ‘Determination of homogeneity and stability of L-Carvone in vehicle’ (Annex 1 of Final report, Study No. 464/17/11HS: L-Carvone – Determination of homogeneity and stability of the application form, VUOS-CETA Report No. 17-742, 2017).
VEHICLE
- Justification for use and choice of vehicle (if other than water): Corn oil
- Amount of vehicle (if gavage): 1 ml/100 g body weight.
- Lot/batch no. (if required): 8002954001 - Analytical verification of doses or concentrations:
- no
- Remarks:
- Homogeneity and stability studies were carried out (see below)
- Details on mating procedure:
- Before mating 2 rats of the same sex were placed in one cage; during the mating period – one male and two females were housed in one cage. After acclimatisation females were mated with males (1 male and 2 females). Vaginal smears were carried out daily in the morning to control fertilization (first time: 24 hours after the first removing to male). Presence of sperms was examined. Day 0 of pregnancy was the day on which sperms in vaginal smears were observed. Pregnant females were randomly distributed to experimental groups. Male rats serve only for mating (they were not administered by the test item and or examined).
- Duration of treatment / exposure:
- DG 5-19
- Frequency of treatment:
- Daily
- Duration of test:
- 20 days
- Dose / conc.:
- 125 mg/kg bw/day (nominal)
- Dose / conc.:
- 250 mg/kg bw/day (nominal)
- Dose / conc.:
- 500 mg/kg bw/day (nominal)
- No. of animals per sex per dose:
- 24 females
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: The dose-range finding experiment (DRFE) with 14-day application period was performed with 4 groups of treated females without control group. The doses selected were 125, 250, 500 and 1000 mg/kg b.w/day.
One female at 1000 mg/kg b.w/day died on the 4th day of study. Four females at 1000 mg/kg b.w/day were humanely killed on the 7th day due to bad health condition and prevention of suffering to the animals. No females died at 125, 250 and 500 mg/kg b.w/day. Health condition control and clinical observations noted the impact of the test substance on the health condition of animals at the dose levels 500 and 1000 mg/kg b.w/day. No changes of animal health status or clinical symptoms of intoxication were observed in females at 125 and 250 mg/kg b.w/day. A significant fall in the body weight of females at 1000 mg/kg b.w/day was recorded on the 7th day of application. Body weight of females at 500 mg/kg b.w/day was slightly decreased compared to 125 and 250 mg/kg b.w/day. Haematological examination did not show significant differences among dose levels except a slight decrease of total leucocyte count in females at 1000 mg/kg b.w/day. No serious macroscopic changes were observed during necropsy in females at 125, 250 and 500 mg/kg b.w/day. Pathological examination of females at 1000 mg/kg b.w/day revealed the following changes: flatulence of stomach and initial autolysis of digestive organs and liver (in one dead female), fur around eyes and nostrils stained by brown-red secretion, decreased thymus, erosions on stomach and forestomach mucosa.
On the basis of the results given above the following dose levels – 125, 250 and 500 mg/kg b.w/day were chosen for the main Prenatal Developmental Toxicity Study. - Maternal examinations:
- DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: All rats were examined for vitality or mortality changes daily during the acclimatization, mating and pregnancy. The health condition was controlled daily during the acclimatization period, during the mating period and during pregnancy. Pre-experimental control of all females was performed to ensure that only the females exhibiting normal behavioral activity would be entered into the study. In administration period this observation was performed before application and immediately after application. During the administration period, clinical observations were made in order to record possible clinical effect of the test item application and all changes in behavior of females. Females were observed in natural conditions in their cages after application - once a day at the similar time each day.
BODY WEIGHT: Yes
- Time schedule for examinations: The body weight of pregnant females was recorded on automatic balances with group mean computing module. First weighing was performed on the 1st day of pregnancy and then on the 5th, 8th, 11th, 14th, 17th and 20th day of pregnancy.
FOOD CONSUMPTION AND COMPOUND INTAKE: Yes; food consumption was determined at three-day intervals; it coincided with the terms of body weight recording.
POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day # 20
- Organs examined: During macroscopic examination, the cranial, thoracic and abdominal cavities were examined and the uterus (incl. the cervix) was removed and weighed. - Ovaries and uterine content:
- The ovaries and uterine content was examined after termination: Yes
Examinations included: In the gravid uterus, the number of viable foetuses, number of dead foetuses, number of early resorptions (implantation without recognizable embryo/foetus) and number of late resorptions (dead embryo or foetus with external degenerative changes) were recorded. The numbers of corpora lutea in both ovaries were noted. Uteri of non-pregnant females were examined to confirm the non-pregnant status. Preimplantation (IUDE (Intra Uterine Death Early)) and post-implantation (IUDL (Intra Uterine Death Late)) losses were calculated from number of implantations (number of foetuses plus number of resorptions), corpora lutea and resorptions. - Fetal examinations:
- Sex and individual body weights of foetuses were recorded. Each foetus was examined for external alterations: symmetry of fore and hind limbs, number of fingers, closing or opening of eye fissures and external auditory canal, symmetry of head, integrity of superior palatum, status of umbilicus and genital papilla were observed.
One half of each litter (one half of female and male foetuses) was examined for soft tissue alterations using careful gross dissection. The second half of each litter was processed and microscopically examined for skeletal and cartilage alterations. Single staining displaying only ossified skeletal structures was used: the foetuses were fixed in ethanol, macerated in potassium hydroxide solution, stained with Alizarin red and placed in glycerine-based solution. The skeletal examination was performed using a stereomicroscope and included examination of skull, clavicle, scapula, sternebra and sternum, ribs, vertebrae, pelvic girdle, forelimb/hindlimb. - Statistics:
- The results statistically significant on probability level 0.05 (p≤0.05) are indicated in the summary tables.
The parametric tests were used for statistical evaluation of:
-body weight of females (5th, 8th, 11th, 14th, 17th, 20th day of pregnancy)
-food consumption (5th, 8th, 11th, 14th, 17th, 20th day of pregnancy)
-corrected body weight (subtraction weight of uterus from surgery body weight of females)
-mean weight of foetuses (males, females, both sex)
-biometry of uteri (absolute and relative weight)
-reimplantation (IUDE) and postimplantation (IUDL) loss
As the first step the test for normality (Shapiro-Wilk test) was performed. If the data were not normally distributed the transformation of data was performed (Box-Cox transformation). If the data were not normal distributed after transformation, the non-parametric tests (Kruskal-Wallis Test and Mann-Whitney test) for comparison of the medians were performed.
If data were normally distributed after transformation, the Variance check (Levene’s test) to verify standard deviations within each group was used. One-Way ANOVA (probability level 0.05) was used to detect whether there were any significant differences amongst the means and then the post hoc statistical testing (Fisher's least significant difference - LSD test) for only statistical significant differences was performed.
The non-parametric tests were used for statistical evaluation of following parameters:
-number of corpora lutea, number of implantations, number of resorptions
-number of live foetuses (males, females, both sex)
-number of dead foetuses
The two-groups Mann-Whitney test (probability level 0.05) was applied.
The categorical data - skeletal foetal findings were analyzed using the generalized linear mixed models with Poisson distribution. - Historical control data:
- Historical control data for foetal skeletal alterations was provided by the lab (2016-2017). Historical control data for foetal skeletal alterations was also available from the Charles River Wistar rat strain (2003-2016).
- Clinical signs:
- effects observed, treatment-related
- Description (incidence and severity):
- No signs of diseases were recorded in control females and females at the lowest and the middle dose levels during the application period. The abdominal position and decreased activity immediately after application were observed in females at the highest dose level in the first and the second week of treatment. Piloerection, red-brown secretion around mouth and salivation were observed sporadically in females at highest dose level from 3rd week to the end of treatment (Table 5).
No clinical changes were recorded in the control females and in the females at the lowest and middle dose level during the whole study. The piloerection was observed in females at the highest dose level during the whole time of clinical observation. Red brown secretion around nostrils was observed sporadically from 2nd week to the end of treatment in females at the highest dose level (Table 6). - Dermal irritation (if dermal study):
- not examined
- Mortality:
- no mortality observed
- Description (incidence):
- There were no unscheduled deaths during the study.
- Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- The body weight of treated females from 8th up to 20th day of pregnancy was statistically significantly decreased at dose level 500 mg/kg b.w./day. The decreased body weight of females at the highest dose level was related to lower body weight increment (Table 3).
Values of corrected body weight (the necropsy body weight of female minus weight of uterus) of females were slightly decreased at the lowest and middle dose levels compared to the control group. The value of corrected body weight was statistically significantly decreased at the highest dose level (Table 8). - Food consumption and compound intake (if feeding study):
- effects observed, treatment-related
- Description (incidence and severity):
- Mean food consumption was significantly decreased compared to control group from 8th day to end of study (correlation with decreased mean of body increment) at the lowest and the highest dose level (Table 4).
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- not examined
- Haematological findings:
- not examined
- Clinical biochemistry findings:
- not examined
- Urinalysis findings:
- not examined
- Behaviour (functional findings):
- not examined
- Immunological findings:
- not examined
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Description (incidence and severity):
- The absolute weight of the uterus was statistically significantly decreased at the highest dose level. The values of relative weight of the uterus in females at all dose levels were similar to the control. Decrease in the absolute weight of the uterus is in accordance with the decrease in body weight of treated females (Table 7).
- Gross pathological findings:
- effects observed, treatment-related
- Description (incidence and severity):
- Pathological findings were observed only in females at the highest dose level: Pathological findings on the spleen – reduced and/or change colour were observed in females at the highest dose level. Kidneys – change of colour was observed in one female. The uterus dilatation (a change probably associated with the physiological oestrous cycle) was detected in one non-pregnant female at the lowest dose level (Table 9).
- Neuropathological findings:
- not examined
- Histopathological findings: non-neoplastic:
- not examined
- Histopathological findings: neoplastic:
- not examined
- Number of abortions:
- no effects observed
- Pre- and post-implantation loss:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Preimplantation losses (Intra Uterine Death Early; IUDE) were increased at the lowest dose level (11.26 %) and slightly decreased at the highest dose level (2.64 %) in comparison with control group (5.92 %). Post-implantation losses (Intra Uterine Death Late; IUDL) were statistically insignificantly increased at the middle dose (15.31 %) in comparison with control (5.98 %) (Table 11).
- Total litter losses by resorption:
- effects observed, treatment-related
- Description (incidence and severity):
- Two females became pregnant and then all implanted conceptus in the uterus were totally resorbed - at the middle dose level (Table 2).
- Early or late resorptions:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Preimplantation losses (Intra Uterine Death Early; IUDE) were increased at the lowest dose level (11.26 %) and slightly decreased at the highest dose level (2.64 %) in comparison with control group (5.92 %). Post-implantation losses (Intra Uterine Death Late ; IUDL) were statistically insignificantly increased at the middle dose (15.31 %) in comparison with control (5.98 %) (Table 11).
- Dead fetuses:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Dead foetuses were found sporadically in control, the highest and in the middle group (1-0-3-2) (Tables 12, 13).
- Changes in pregnancy duration:
- not specified
- Changes in number of pregnant:
- effects observed, treatment-related
- Description (incidence and severity):
- The number of non-pregnant females after mating was 1-4-2-8 in the respective dose groups. The number of treated females with live foetuses on the 20th day of pregnancy was the lowest at the highest dose level: 23-20-20-16 (Table 2).
- Other effects:
- not examined
- Dose descriptor:
- NOAEL
- Effect level:
- 250 mg/kg bw/day (nominal)
- Basis for effect level:
- body weight and weight gain
- gross pathology
- Abnormalities:
- no effects observed
- Fetal body weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- The mean body weight of foetuses was statistically significantly decreased at the highest dose level compared to the control group. At the lowest and at the middle dose level the mean body weight of foetuses was similar to the control (Table 14).
- Reduction in number of live offspring:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Statistically significant differences in the number of live foetuses, live male foetuses and live female foetuses were not found at any dose level. The total number of live fetuses in groups was decreased at the lowest and the highest dose level compared to the control group, but the average total number of fetuses per litter was well balanced with control. (Tables 12, 13).
- Changes in sex ratio:
- no effects observed
- Description (incidence and severity):
- Statistically significant differences in the number of live male foetuses or live female foetuses were not found at any dose level (Tables 12, 13).
- Changes in litter size and weights:
- no effects observed
- Description (incidence and severity):
- Statistically significant differences in the number of live foetuses were not found at any dose level (Table 13).
- Description (incidence and severity):
- See skeletal malformations below.
- External malformations:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Examination of symmetry of fore and hind limbs, number of fingers, closing or opening of eye fissures and external auditory canal, symmetry of head, integrity of superior palatum, status of umbilicus and genital papilla were performed. No external changes were recorded (Table 15). Dead foetuses were recorded in control, middle and in the highest dose levels. One foetus without tail was observed in the control group and one in the middle and the lowest dose levels.
- Skeletal malformations:
- effects observed, treatment-related
- Description (incidence and severity):
- During the examination of foetuses, cranium incomplete ossification was found in all groups (including control). The occurrence of incomplete ossification of frontal, interparietal, parietal, palatinum bone and tympanic annulus was the highest in the control group. On the contrary, the evidence of incomplete ossification of the supraoccipital bone was increased in females at the highest dose level (100 % of litters) in comparison with control group (56.52 % of litters).
Bipartite ossification of the supraoccipital bone (40% of litters) and unossified supraoccipital bone (20% of litters) was found only at the highest dose level. The increased portion of litters with holes in the supraoccipital bone was observed in treated groups in comparison to the control group (47.83 % - 72.22 % - 60.00 % - 80.00 %). The malformation of tympanic annulus (short) was found in 13.33 % of litters only at the highest dose level.
During the examination of the foetal skeleton, a decreased number of ossification sites of the sternum was recorded in foetuses in all groups. The ossification sites were either incompletely ossified or unossified. The evidence of incomplete ossification of ossification sites was the same in all groups including the control group. The evidence of bipartite ossification was increased in foetuses at the middle dose level. The evidence of unossified ossification sites were statistically significantly increased at the highest dose level (Table 21). Asymmetric ossification of sternebra was found only in foetuses of treated females (0.00 % - 16.67 % - 10.00 % - 13.33 %).
Anomalies of vertebrae and ribs were observed. The examination of vertebrae revealed mainly dumbbell and bipartite ossification of vertebrae (thoracic centrum or lumbar centrum). The incidence of dumbbell ossification was higher in treated groups (8.70 % - 16.67 % - 25.00 % - 13.33 %) in comparison with controls. The portions of litters with bipartite ossification of vertebrae were increased in the middle and the highest dose level in comparison with the control group (13.04 % - 11.11 % -20.00 % - 20.00 %).
The malformations of vertebrae (absent vertebrae in thoracic, lumbar or sacral centrum) were observed in 11.11 % of litters at the lowest dose level and in 5.00 % of litters at the middle dose level.
Ribs – supernumerary sites were observed in all groups with the highest occurrence in the control group and in the lowest dose level group. Malformations of ribs (absent) were observed only in treated group litters (0.00 % - 16.67 % - 10.00 % - 20.00 %).
Other changes of foetus skeleton were found without treatment relation.
Refer to tables 17-21. - Visceral malformations:
- no effects observed
- Description (incidence and severity):
- No visceral malformation findings were found at all dose levels and in the control group (Table 16).
- Other effects:
- not specified
- Key result
- Dose descriptor:
- LOAEL
- Effect level:
- ca. 125 mg/kg bw/day (nominal)
- Sex:
- male/female
- Basis for effect level:
- skeletal malformations
- Dose descriptor:
- NOAEL
- Effect level:
- 125 mg/kg bw/day (nominal)
- Sex:
- male/female
- Basis for effect level:
- other: transitional findings of supraooccipitale bone, sternabra, vertebrae and malformations of the ribs
- Abnormalities:
- effects observed, treatment-related
- Localisation:
- other: transitional findings of supraooccipital bone, sternabra, vertebrae and malformations of the ribs
- Developmental effects observed:
- yes
- Lowest effective dose / conc.:
- 125 mg/kg bw/day (nominal)
- Treatment related:
- yes
- Relation to maternal toxicity:
- developmental effects in the absence of maternal toxicity effects
- Dose response relationship:
- yes
- Relevant for humans:
- not specified
- Conclusions:
- In a pre-natal developmental toxicity study in rats, the NOAEL for toxicity in pregnant females was established 250 mg/kg b.w./day. This NOAEL value is based on no occurrence of mortality, no changes in health condition status, no reproductive-related pathological findings and no changes in reproduction parameters at the dose 250 mg/kg b.w./day.
The NOAEL for prenatal development is < 125 mg/kg b.w./day. This NOAEL value is based on the occurrence of transitional findings of supraooccipital bone, sternabrae, vertebrae and malformations of the ribs at a dose of 125 mg/ kg b.w./day. The dose level 125 mg/kg b.w./day can be treated as the LOAEL in this study. - Executive summary:
In a pre-natal developmental toxicity study (18-324), L-carvone (99.16%) was administered to 24 female Wistar CRL rats in corn oil by gavage at dose levels of 0, 125, 250 and 500 mg/kg b.w./day from days 5 through 19 of gestation.
There were no unscheduled mortalities during the study. A negative effect of test item treatment on the growth of maternal animals was observed at the highest dose 500 mg/kg b.w./day. The body weight was statistically significantly decreased in this group and this correlated with the statistically significantly decrease of food consumption. Adverse changes of health condition and clinical symptoms of intoxication were observed only in females at the dose 500 mg/kg b.w./day. Also, serious pathological findings and a statistically significant decrease in the absolute weight of the uterus were revealed at this dose level. These findings were clearly related to treatment by the test item. There were no changes in reproduction parameters in treated groups.
The examination of foetuses revealed a statistically significant decrease in the mean body weights at the dose 500 mg/kg b.w./day. The decreased weight of foetuses could be associated with a negative influence of the test item on the growth of maternal animals.
Examination of foetal skull and skeleton revealed anomalies in all treated groups. The incomplete ossification of the supraoccipital bone, could be associated with lower foetal body weight, especially at the highest dose level. Equally, transitional findings in the foetal skull, such as bipartite ossification of the supraoccipital bone and unossified supraoccipital bone could be associated with maternal toxicity, due to incidence at the dose 500 mg/kg b.w./day only. The increased incidence of transitional findings in all treated groups in comparison with control group – mainly hole in supraoccipital bone, asymmetric ossification of sternebra and bipartite and dumbbell ossification of vertebrae could be considered as adverse effect of the test item to foetuses, because no maternal toxicity was observed at the 125 and 250 mg/kg b.w./day dose levels. The malformations of ribs (absent) were observed only in treated groups.
The increased incidence of transitional findings (in foetuses of treated females compared to control females) could be considered as an adverse effect of the test item to early prenatal development of organism in uterus. The effect of the test item on the incidence of malformation of ribs in treated groups is disputable.
In a pre-natal developmental toxicity study in rats, the NOAEL for toxicity in pregnant females was established 250 mg/kg b.w./day. This NOAEL value is based on no occurrence of mortality, no changes in health condition status, no reproductive-related pathological findings and no changes in reproduction parameters at the dose 250 mg/kg b.w./day.
The NOAEL for prenatal development is < 125 mg/kg b.w./day. This NOAEL value is based on the occurrence of transitional findings of supraooccipital bone, sternabrae, vertebrae and malformations of the ribs at a dose of 125 mg/ kg b.w./day. The dose level 125 mg/kg b.w./day can be treated as the LOAEL in this study.
Reference
Tables from the main study report
Table 2: Pregnancy results
Pregnancy results |
|||
Group Code |
Number of females with live foetuses |
Numbers of females without foetuses and implantations* |
Number of females without live foetuses but with implantation andresorption |
0 |
23 |
1(109) |
- |
125 |
20 |
4(130, 132, 137, 141) |
- |
250 |
20 |
2(162, 167) |
2(165,172) |
500 |
16 |
8(178,179,184,186,188,189,190,191) |
- |
Note:
* numbers in parentheses = individual labels of single animals
- Only the data from females with live foetuses were used for calculations of means.
- Data from females with uterus implantations were used for calculation of preimplantation (IUDE) and post-implantation (IUDL) losses.
Table 3: Body weight in grams (mean±standard deviation)
Body weight in grams(mean±standard deviation) |
||||
Day of pregnancy |
Group Code |
|||
0 |
125 |
250 |
500 |
|
1stday |
280.8 ± 13.6 |
278.2 ± 12.5 |
282.2 ± 13.5 |
275.4 ± 12.6 |
5thday |
295.5 ± 16.1 |
294.1 ± 14.2 |
302.2 ± 12.5 |
291.3 ± 13.8 |
8thday |
305.5 ± 15.6 |
300.9 ± 15.1 |
307.8 ± 11.6 |
286.0 ± 16.3 |
11thday |
324.0 ± 18.7 |
317.8 ± 17.1 |
327.3 ± 14.8 |
294.1 ± 18.0 |
14thday |
341.1 ± 19.3 |
337.9 ± 19.1 |
345.9 ± 15.6 |
305.9 ± 18.9 |
17thday |
377.5 ± 23.5 |
374.0 ± 22.0 |
383.2 ± 20.6 |
328.5 ± 22.2 |
20thday |
433.5 ± 29.5 |
424.1 ± 27.5 |
431.0 ± 26.7 |
338.0 ± 29.1 |
Mean increment |
152.8 ± 20.7 |
145.9 ± 21.1 |
148.9 ± 18.5 |
62.5 ± 30.0 |
Note:Values statistically significant on probability level 0.05 (p≤0.05)are shaded.
Table 4: Food consumption g/animal/day (mean)
Food consumption g/animal/day(mean) |
||||
Day of pregnancy |
Group Code |
|||
0 |
125 |
250 |
500 |
|
5thday |
22.82 |
22.65 |
24.62 |
22.92 |
8thday |
19.80 |
16.88 |
17.36 |
12.53 |
11thday |
23.35 |
20.55 |
21.70 |
19.02 |
14thday |
23.42 |
21.83 |
23.60 |
19.38 |
17thday |
26.01 |
23.58 |
25.10 |
18.98 |
20thday |
28.36 |
24.34 |
26.73 |
12.78 |
Table 5: Health condition control
Health condition control |
||||
Week of pregnancy |
Group Code |
|||
0 |
125 |
250 |
500 |
|
1stweek
|
1 |
1 |
1 |
2 – 11 |
2ndweek
|
1 |
1 |
1 |
2 – 9 |
3rdweek
|
1 |
1 6 – 1 |
1 |
3 – 1 4 – 2 5 – 2 |
Note:Type of finding – number of females affected.1– physiological appearance,2–abdominal position and decreased activity immediately after treatment,3– piloerection,4– red-brown secretion around mouth,5– salivation, 6 – tumour in mammary gland
Table 6: Clinical observation
Clinical observation |
||||
Week of pregnancy |
Group Code |
|||
0 |
125 |
250 |
500 |
|
1stweek |
1 |
1 |
1 |
2 – 6 |
2ndweek |
1 |
1 |
1 |
2 – 4 3 – 1 |
3rdweek |
1 |
1 |
1 |
2 – 2 3 – 2 |
Note:Type of finding – number of females affected. 1– no clinical symptoms of intoxication,2– piloerection,3– red-brown secretion around nostrils
Table 7: Biometry of uterus (mean±standard deviation)
Biometry of uterus(mean±standard deviation) |
||||
Parameter |
Group Code |
|||
0 |
125 |
250 |
500 |
|
Mean necropsy body weight of females (g) |
433.5 ± 29.5 |
424.1 ± 27.5 |
431.0 ± 26.7 |
338.0 ± 29.1 |
Mean absolute weight of uterus (g) |
87.7878 ± 14.0922 |
83.5355 ± 18.6988 |
91.4955 ± 12.4892 |
71.3027 ± 12.4658 |
Mean relative weight of uterus (%) |
20.2332 ± 2.8762 |
19.6477 ± 4.1573 |
21.2230 ± 2.5682 |
21.0298 ± 2.6623 |
Note:Values statistically significant on probability level 0.05(p<0.05) are shaded.
Table 8: Body weight – corrected* (mean±standard deviation)
Body weight - corrected *(mean±standard deviation) |
||||
Parameter |
Group Code |
|||
0 |
125 |
250 |
500 |
|
Body weight(g) |
345.8 ± 25.5 |
340.5 ± 25.2 |
339.5 ± 23.2 |
266.7 ± 21.6 |
Note:Values statistically significant on probability level 0.05(p<0.05) are shaded.
*body weight correction = necropsy body weight of female – weight of uterus
Table 9: Macroscopic findings(number of females with pathological findings)
Macroscopic findings(number of females with pathological findings) |
||||
Parameter |
Group Code |
|||
0 |
125 |
250 |
500 |
|
Number of examined females |
24 |
24 |
24 |
24 |
Number of dead females |
0 |
0 |
0 |
0 |
Without pathological findings |
24 |
24 |
24 |
18 |
Spleen:reduced |
0 |
0 |
0 |
4 |
Spleen:light brown-orange colour |
0 |
0 |
0 |
3 |
Kidneys:light brown-orange colour |
0 |
0 |
0 |
1 |
Uterus:dilatation |
0 |
1 |
0 |
0 |
Table 10: Parameters of reproduction (number per female, mean±SD)
Parameters of reproduction(number per female, mean±SD) |
||||
Parameter |
Group Code |
|||
0 |
125 |
250 |
500 |
|
implantations |
16.09 ± 1.88 |
14.90 ± 3.54 |
15.59 ± 4.56 |
16.63 ± 2.33 |
resorptions |
0.91 ± 1.56 |
0.50 ± 0.83 |
1.27 ± 1.39 |
0.50 ± 0.82 |
corpora lutea |
17.17 ± 1.99 |
16.65 ± 1.50 |
16.59 ± 4.63 |
17.00 ± 1.55 |
Note: Statistically significant differences on probability level 0.05(p<0.05)were not detected.
Table 11: IUDE and IUDL(% per female, mean ±SD)
I U D E and I U D L(% per female, mean ±SD) |
||||
Parameter |
Group Code |
|||
0 |
125 |
250 |
500 |
|
IUDE |
5.92± 9.30 |
11.26 ± 17.77 |
6.25 ± 6.56 |
2.64 ± 7.75 |
IUDL |
5.98 ± 10.36 |
2.86 ± 4.72 |
15.31 ± 28.41 |
2.84 ± 4.62 |
Note: Statistically significant differences on probability level 0.05(p<0.05)were not detected.
The mean of preimplantation (IUDE) and post-implantation losses (IUDL) were calculated from individual data of females.
Table 12: Number of foetuses
Number of foetuses (total in group) |
||||
Parameter |
Group Code |
|||
0 |
125 |
250 |
500 |
|
Total number of live foetuses |
349 |
288 |
315 |
258 |
Number of live foetuses – males |
164 |
149 |
154 |
135 |
Number of live foetuses – females |
185 |
139 |
161 |
123 |
Number of dead foetuses |
1 |
0 |
3 |
2 |
Table 13: Number of foetuses (per litter; mean ± SD)
Number of foetuses (per litter; mean ± SD) |
||||
Parameter |
Group Code |
|||
0 |
125 |
250 |
500 |
|
Total number of live foetuses |
15.17 ± 2.69 |
14.40 ± 3.22 |
14.32 ± 5.07 |
16.13 ± 2.22 |
Number of live foetuses – males |
7.13 ± 2.12 |
7.45 ± 2.67 |
7.00 ± 3.19 |
8.44 ± 2.87 |
Number of live foetuses – females |
8.04 ± 2.38 |
6.95 ± 1.85 |
7.32 ± 2.95 |
7.69 ± 2.44 |
Number of dead foetuses |
0.04 ± 0.21 |
0.00 ± 0.00 |
0.14 ± 0.47 |
0.13 ± 0.34 |
Note: Statistically significant differences on probability level 0.05(p<0.05)were not detected.
Table 14:Body weight of foetuses (grams, mean ± SD)
Body weight of foetuses (grams, mean ± SD) |
||||
Parameter |
Group Code |
|||
0 |
125 |
250 |
500 |
|
mean weight of all foetus |
3.94074 ± 0.52895 |
3.96180 ± 1.08688 |
3.97550 ± 0.44793 | 2.73026 ± 0.43264 |
mean weight of male foetus |
4.05726 ± 0.56480 |
4.02498 ± 1.07858 |
4.05257 ± 0.45354 |
2.77061 ± 0.45029 |
mean weight of female foetus |
3.83062 ± 0.49848 |
3.89160 ± 1.10542 |
3.89968 ± 0.46388 |
2.69040 ± 0.44140 |
Note:Values statistically significant on probability level 0.05 (p<0.05) areshaded.
Table 15: External alterations - foetuses
External alterations - foetuses |
||||
Alteration |
Group Code |
|||
0 |
125 |
250 |
500 |
|
Total number of examined foetuses |
350 |
288 |
318 |
260 |
Total number of examined litters |
23 |
20 |
22 |
16 |
With pathological findings(number of affected foetuses) |
1 |
1 |
4 |
2 |
Number of examined foetuses in litter (mean ± SD) |
15.20±2.66 |
14.40±3.22 |
14.45±4.68 |
16.25±2.29 |
Total number of foetuses with alteration |
1 |
1 |
4 |
2 |
Dead foetus - without tail |
1 |
0 |
0 |
0 |
Live foetus – without tail |
0 |
1 |
1 |
0 |
Dead foetus |
0 |
0 |
3 |
2 |
Number of foetuses with alteration in litter (mean ± SD) |
0.04±0.21 |
0.05±0.22 |
0.18±0.50 |
0.13±0.34 |
Portion of foetuses with alteration in litter (% mean ± SD) |
0.32±1.52 |
0.33±1.49 |
9.38±29.36 |
0.70±1.91 |
Table 16: Macroscopic changes of soft tissues - foetuses
Macroscopic changes of soft tissues – foetuses |
||||
Alteration |
Group Code |
|||
0 |
125 |
250 |
500 |
|
Total number of examined foetuses |
350 |
288 |
318 |
260 |
Total number of examined litters |
23 |
20 |
22 |
16 |
With pathological findings - total(number of affected foetuses) |
0 |
0 |
0 |
0 |
Number of examined foetuses in litter (mean ± SD) |
15.20± 2.66 |
14.40± 3.22 |
14.45± 4.68 |
16.25± 2.29 |
Total number of foetuses with alteration |
0 |
0 |
0 |
0 |
Number of foetuses with alteration in litter (mean ± SD) |
0.00 ±0.00 |
0.00 ±0.00 |
0.00 ±0.00 |
0.00 ±0.00 |
Portion of foetuses with alteration in litter (% mean± SD ) |
0.00
|
0.00
|
0.00
|
0.00
|
Table 17: Skeletal alterations(number of affected foetuses)
Skeletal alterations(number of affected foetuses) |
|||||||
|
0 |
125 |
250 |
500 |
|||
Number of examined foetuses |
184 | 135 |
170 | 126 |
|||
CRANIUM |
Frontal bone – i.o. |
5 |
1 |
0 |
0 |
||
Interparietal bone – i.o. |
17 |
7 |
13 |
7 |
|||
Premaxilla –short** |
0 |
1 |
0 |
0 |
|||
Mandible – i.o. |
1 |
0 |
0 |
0 |
|||
Mandible – short** |
0 |
1 |
0 |
0 |
|||
Palatinum bone – i.o. |
9 |
5 |
0 |
2 |
|||
Parietal bone – i.o. |
20 |
4 |
7 |
8 |
|||
Supraoccipital bone – i.o. |
24 |
9 |
15 |
32 |
|||
Supraoccipital bone – u. |
0 |
0 |
0 |
3 |
|||
Supraoccipital bone – hole |
19 |
19 |
20 |
28 |
|||
Supraoccipital bone – b.o. |
0 |
0 |
0 |
14 |
|||
Tympanic annulus – i.o. |
6 |
6 |
4 |
1 |
|||
Tympanic annulus – short** |
0 |
0 |
0 |
3 |
|||
Arcus zygomaticus – misshapen** |
0 |
1 |
0 |
0 |
|||
Basisphenoid –i.o. |
0 |
0 |
1 |
0 |
|||
Squamous part of temporal bone –i.o. |
0 |
0 |
0 |
1 |
|||
STERNEBRA |
Sternebra – decreased number of ossification sites* |
175 |
126 |
159 |
126 |
||
Sternebrae – ossification sites i.o. |
169 |
122 |
153 |
104 |
|||
Sternebrae – ossification sites u. |
76 | 68 |
92 | 111 |
|||
Sternebrae – ossification sites b.o. |
5 |
0 |
15 |
4 |
|||
Sternebrae – ossification sites - asymmetric ossification |
0 |
4 |
2 |
2 |
|||
Sternebrae – ossification sites - absent** |
0 |
1 |
0 |
0 |
|||
Sternebrae – ossification sites – malpositioned** |
1 |
0 |
0 |
0 |
|||
VERTEBRAE |
Vertebrae - thorac, lumb. centrum – d.o. |
2 |
3 |
5 |
4 |
||
Vertebrae - thoracic centrum – b.o. |
4 |
4 |
6 |
6 |
|||
Vertebrae – thorac., lumb., sacr., - absent** |
0 |
2 |
1 |
0 |
|||
RIBS |
Ribs – wavy |
5 |
5 |
2 |
0 |
||
Ribs – i.o. |
1 |
0 |
0 |
0 |
|||
Ribs – supernumerary site |
17 |
17 |
11 |
7 |
|||
Ribs – short |
0 |
1 |
0 |
0 |
|||
Ribs – absent** |
0 |
3 |
3 |
3 |
|||
Humerus – short** |
0 |
1 |
0 |
0 |
|||
Scapula – i.o. |
8 |
4 |
0 |
0 |
|||
Ilium – i.o. |
1 |
1 |
0 |
1 |
|||
Pubis– i.o. |
0 |
1 |
0 |
5 |
|||
Ischium – i.o. |
1 |
1 |
0 |
11 |
|||
Note for tables 17-20:
i.o. – incomplete ossification
u. – unossified
d.o. – dumbbell ossification
b.o. – bipartite ossification
* - the mentioned ossification sites are either incompletly ossified or unossified
** - malformation
The transitional findings that are included to the “grey zone anomalies”are shaded
Table 18: Skeletal alterations(number of litters with affected foetuses)
Skeletal alterations(number of litters with affected foetuses) |
|||||||
|
0 |
125 |
250 |
500 |
|||
Number of examined litters |
23 | 18 |
20 | 15 |
|||
CRANIUM |
Frontal bone – i.o. |
3 |
1 |
0 |
0 |
||
Interparietal bone – i.o. |
12 |
5 |
8 |
5 |
|||
Premaxilla –short** |
0 |
1 |
0 |
0 |
|||
Mandible – i.o. |
1 |
0 |
0 |
0 |
|||
Mandible – short** |
0 |
1 |
0 |
0 |
|||
Palatinum bone – i.o. |
6 |
2 |
0 |
2 |
|||
Parietal bone – i.o. |
13 |
2 |
5 |
5 |
|||
Supraoccipital bone – i.o. |
13 |
6 |
9 |
15 |
|||
Supraoccipital bone – u. |
0 |
0 |
0 |
3 |
|||
Supraoccipital bone – hole |
11 |
13 |
12 |
12 |
|||
Supraoccipital bone – b.o. |
0 |
0 |
0 |
6 |
|||
Tympanic annulus – i.o. |
5 |
3 |
4 |
1 |
|||
Tympanic annulus – short** |
0 |
0 |
0 |
2 |
|||
Arcus zygomaticus – misshapen** |
0 |
1 |
0 |
0 |
|||
Basisphenoid –i.o. |
0 |
0 |
1 |
0 |
|||
Squamous part of temporal bone –i.o. |
0 |
0 |
0 |
1 |
|||
STERNEBRA |
Sternebrae – decreased number of ossification sites * |
23 |
18 |
20 |
15 |
||
Sternebrae – ossification sites i.o. |
23 |
18 |
20 |
15 |
|||
Sternebrae – ossification sites u. |
18 | 14 |
17 | 14 |
|||
Sternebrae – ossification sites b.o. |
4 |
0 |
6 |
3 |
|||
Sternebrae – ossification sites - asymmetric ossification |
0 |
3 |
2 |
2 |
|||
Sternebrae – ossification sites - absent** |
0 |
1 |
0 |
0 |
|||
Sternebrae – ossification sites – malpositioned** |
1 |
0 |
0 |
0 |
|||
VERTEBRAE |
Vertebrae - thorac, lumb. centrum – d.o. |
2 |
3 |
5 |
2 |
||
Vertebrae - thoracic centrum – b.o. |
3 |
2 |
4 |
3 |
|||
Vertebrae – thorac., lumb., sacr., - absent** |
0 |
2 |
1 |
0 |
|||
RIBS |
Ribs – wavy |
3 |
2 |
1 |
0 |
||
Ribs – i.o. |
1 |
0 |
0 |
0 |
|||
Ribs – supernumerary site |
11 |
11 |
7 |
4 |
|||
Ribs – short |
0 |
1 |
0 |
0 |
|||
Ribs – absent** |
0 |
3 |
2 |
3 |
|||
Humerus – short** |
0 |
1 |
0 |
0 |
|||
Scapula – i.o. |
5 |
3 |
0 |
0 |
|||
Ilium – i.o. |
1 |
1 |
0 |
1 |
|||
Pubis– i.o. |
0 |
1 |
0 |
2 |
|||
Ischium – i.o. |
1 |
1 |
0 |
4 |
|||
Table 19: Skeletal alterations (% portion of affected foetuses in litter(mean ± SD)
Skeletal alterations(% portion of affected foetuses in litter (mean ± SD)) |
|||||||
|
0 |
125 |
250 |
500 |
|||
Number of examined foetuses |
184 | 135 |
170 | 126 |
|||
Number of examined litters |
23 | 18 |
20 | 15 |
|||
CRANIUM |
Frontal bone – i.o. |
3.44 ± 11.00 |
0.93 ± 3.93 |
0.00 ± 0.00 |
0.00 ± 0.00 |
||
Interparietal bone – i.o. |
10.26 ± 14.79 |
5.17 ± 9.88 |
7.82 ± 10.97 |
6.95 ±11.87 |
|||
Premaxilla –short** |
0.00 ± 0.00 |
0.69 ± 2.95 |
0.00 ± 0.00 |
0.00 ± 0.00 |
|||
Mandible – i.o. |
0.48 ± 2.32 |
0.00 ± 0.00 |
0.00 ± 0.00 |
0.00 ± 0.00 |
|||
Mandible – short** |
0.00 ± 0.00 |
0.69 ± 2.95 |
0.00 ± 0.00 |
0.00 ± 0.00 |
|||
Palatinum bone – i.o. |
5.07 ± 9.77 |
6.35 ± 23.61 |
0.00 ± 0.00 |
1.85 ± 5.00 |
|||
Parietal bone – i.o. |
10.98 ±13.34 |
3.47 ± 11.98 |
4.17 ± 7.97 |
6.66 ± 12.36 |
|||
Supraoccipital bone – i.o. |
12.76 ±14.04 |
6.50 ± 10.44 |
8.93 ± 14.43 |
26.66 ± 17.59 |
|||
Supraoccipital bone – u. |
0.00 ± 0.00 |
0.00 ± 0.00 |
0.00 ± 0.00 |
2.59 ± 5.50 |
|||
Supraoccipital bone – hole |
9.16 ± 11.16 |
15.08 ± 15.25 |
11.85 ± 14.25 |
22.92 ± 18.80 |
|||
Supraoccipital bone – b.o. |
0.00 ± 0.00 |
0.00 ± 0.00 |
0.00 ± 0.00 |
12.37 ± 19.58 |
|||
Tympanic annulus – i.o. |
3.83 ± 8.35 |
6.87 ± 23.55 |
2.54 ± 5.26 |
0.67 ± 2.58 |
|||
Tympanic annulus – short** |
0.00 ± 0.00 |
0.00 ± 0.00 |
0.00 ± 0.00 |
2.07 ± 5.73 |
|||
Arcus zygomaticus – misshapen** |
0.00 ± 0.00 |
0.93 ± 3.93 |
0.00 ± 0.00 |
0.00 ± 0.00 |
|||
Basisphenoid –i.o. |
0.00 ± 0.00 |
0.00 ± 0.00 |
0.56 ± 2.48 |
0.00 ± 0.00 |
|||
Squamous part of temporal bone –i.o. |
0.00 ± 0.00 |
0.00 ± 0.00 |
0.00 ± 0.00 |
0.74 ± 2.87
|
|||
STERNEBRA |
Sternebrae – decreased number of ossification sites* |
94.30 ± 13.34 |
91.45 ± 19.10 |
94.74 ± 20.41 |
100.00 ± 0.00 |
||
Sternebrae – ossification sites i.o. |
90.96 ± 13.13 |
88.67 ± 19.01 |
91.30 ± 20.83 |
82.55 ± 20.66 |
|||
Sternebrae – ossification sites u. |
41.56 ± 32.24 |
48.70 ± 35.04 |
53.63 ± 31.72 |
88.89 ± 29.99 |
|||
Sternebrae – ossification sites b.o. |
3.16 ± 7.37 |
0.00 ± 0.00 |
8.78 ± 14.76 |
2.89 ± 6.50 |
|||
Sternebrae – ossification sites - asymmetric ossification |
0.00 ± 0.00 |
2.70 ± 6.75 |
1.18 ± 3.64 |
1.48 ± 3.91 |
|||
Sternebrae – ossification sites - absent** |
0.00 ± 0.00 |
0.69 ± 2.95 |
0.00 ± 0.00 |
0.00 ± 0.00 |
|||
Sternebrae – ossification sites – malpositioned** |
0.54 ± 2.61 |
0.00 ± 0.00 |
0.00 ± 0.00 |
0.00 ± 0.00 |
|||
VERTEBRAE |
Vertebrae - thorac, lumb. centrum – d.o. |
0.97 ± 3.20 |
2.01 ± 4.62 |
2.67 ± 4.75 |
2.96 ± 7.82 |
||
Vertebrae - thoracic centrum – b.o. |
1.99 ± 5.57 |
2.55 ± 8.23 |
4.07 ± 10.29 |
4.44 ± 11.73 |
|||
Vertebrae – thorac., lumb., sacr., - absent** |
0.00 ± 0.00 |
1.39 ± 4.04 |
0.56 ± 2.48 |
0.00 ± 0.00 |
|||
RIBS |
Ribs – wavy |
3.23 ± 9.15 |
3.16 ± 10.71 |
1.25 ± 5.59 |
0.00 ± 0.00 |
||
Ribs – i.o. |
0.72 ± 3.48 |
0.00 ± 0.00 |
0.00 ± 0.00 |
0.00 ± 0.00 |
|||
Ribs – supernumerary site |
8.66 ± 10.86 |
12.98 ± 14.83 |
6.21 ± 10.01 |
5.19 ± 10.17 |
|||
Ribs – short |
0.00 ± 0.00 |
0.69 ± 2.95 |
0.00 ± 0.00 |
0.00 ± 0.00 |
|||
Ribs – absent** |
0.00 ± 0.00 |
2.01 ± 4.62 |
1.88 ± 6.60 |
2.53 ±5.26 |
|||
Humerus – short** |
0.00 ± 0.00 |
0.56 ± 2.36 |
0.00 ± 0.00 |
0.00 ± 0.00 |
|||
Scapula – i.o. |
4.66 ± 10.33 |
2.70 ± 6.75 |
0.00 ± 0.00 |
0.00 ± 0.00 |
|||
Ilium – i.o. |
0.87 ± 4.17 |
0.69 ± 2.95 |
0.00 ± 0.00 |
0.74 ± 2.87 |
|||
Pubis– i.o. |
0.00 ± 0.00 |
0.69 ± 2.95 |
0.00 ± 0.00 |
5.40 ± 17.34 |
|||
Ischium – i.o. |
0.48 ± 2.32 |
0.69 ± 2.95 |
0.00 ± 0.00 |
10.58 ± 26.40 |
|||
Table 20: Skeletal alterations (% portion of litters with affected foetuses)
Skeletal alterations(% portion of litters with affected foetuses) |
|||||||
|
0 |
125 |
250 |
500 |
|||
Number of examined litters |
23 | 18 |
20 | 15 |
|||
CRANIUM |
Frontal bone – i.o. |
13.04 |
5.56 |
0.00 |
0.00 |
||
Interparietal bone – i.o. |
52.17 |
27.78 |
40.00 |
33.33 |
|||
Premaxilla –short** |
0.00 |
5.56 |
0.00 |
0.00 |
|||
Mandible – i.o. |
4.35 |
0.00 |
0.00 |
0.00 |
|||
Mandible – short** |
0.00 |
5.56 |
0.00 |
0.00 |
|||
Palatinum bone – i.o. |
26.09 |
11.11 |
0.00 |
13.33 |
|||
Parietal bone – i.o. |
56.52 |
11.11 |
25.00 |
33.33 |
|||
Supraoccipital bone – i.o. |
56.52 |
33.33 |
45.00 |
100.00 |
|||
Supraoccipital bone – u. |
0.00 |
0.00 |
0.00 |
20.00 |
|||
Supraoccipital bone – hole |
47.83 |
72.22 |
60.00 |
80.00 |
|||
Supraoccipital bone – b.o. |
0.00 |
0.00 |
0.00 |
40.00 |
|||
Tympanic annulus – i.o. |
21.74 |
16.67 |
20.00 |
6.67 |
|||
Tympanic annulus – short** |
0.00 |
0.00 |
0.00 |
13.33 |
|||
Arcus zygomaticus – misshapen** |
0.00 |
5.56 |
0.00 |
0.00 |
|||
Basisphenoid –i.o. |
0.00 |
0.00 |
5.00 |
0.00 |
|||
Squamous part of temporal bone –i.o. |
0.00 |
0.00 |
0.00 |
6.67 |
|||
STERNEBRA |
Sternebra – decreased number of ossification sites* |
100.00 |
100.00 |
100.00 |
100.00 |
||
Sternebrae – ossification sites i.o. |
100.00 |
100.00 |
100.00 |
100.00 |
|||
Sternebrae – ossification sites u. |
78.26 |
77.78 |
85.00 |
93.33 |
|||
Sternebrae – ossification sites b.o. |
17.39 |
0.00 |
30.00 |
20.00 |
|||
Sternebrae – ossification sites - asymmetric ossification |
0.00 |
16.67 |
10.00 |
13.33 |
|||
Sternebrae – ossification sites - absent** |
0.00 |
5.56 |
0.00 |
0.00 |
|||
Sternebrae – ossification sites – malpositioned** |
4.35 |
0.00 |
0.00 |
0.00 |
|||
VERTEBRAE |
Vertebrae - thorac, lumb. centrum – d.o. |
8.70 |
16.67 |
25.00 |
13.33 |
||
Vertebrae - thoracic centrum – b.o. |
13.04 |
11.11 |
20.00 |
20.00 |
|||
Vertebrae – thorac., lumb., sacr., - absent** |
0.00 |
11.11 |
5.00 |
0.00 |
|||
RIBS |
Ribs – wavy |
13.04 |
11.11 |
5.00 |
0.00 |
||
Ribs – i.o. |
4.35 |
0.00 |
0.00 |
0.00 |
|||
Ribs – supernumerary site |
47.83 |
61.11 |
35.00 |
26.67 |
|||
Ribs – short |
0.00 |
5.56 |
0.00 |
0.00 |
|||
Ribs – absent** |
0.00 |
16.67 |
10.00 |
20.00 |
|||
Humerus – short** |
0.00 |
5.56 |
0.00 |
0.00 |
|||
Scapula – i.o. |
21.74 |
16.67 |
0.00 |
0.00 |
|||
Ilium – i.o. |
4.35 |
5.56 |
0.00 |
6.67 |
|||
Pubis– i.o. |
0.00 |
5.56 |
0.00 |
13.33 |
|||
Ischium – i.o. |
4.35 |
5.56 |
0.00 |
26.67 |
|||
Table 21: Skeletal alterations–STATISTICAL ANALYSIS
Skeletal alterations(number of affected foetuses) |
|||||
|
0 |
125 |
250 |
500 |
|
Number of examined foetuses |
184 |
135 |
170 |
126 |
|
CRANIUM |
Frontal bone – i.o. |
5 |
1 |
0 |
0 |
Interparietal bone – i.o. |
17 |
7 |
13 |
7 |
|
Premaxilla –short** |
0 |
1 |
0 |
0 |
|
Mandible – i.o. |
1 |
0 |
0 |
0 |
|
Mandible – short** |
0 |
1 |
0 |
0 |
|
Palatinum bone – i.o. |
9 |
5 |
0 |
2 |
|
Parietal bone – i.o. |
20 |
4 |
7 |
8 |
|
Supraoccipital bone – i.o. |
24 |
9 |
15 |
32 |
|
Supraoccipital bone – u. |
0 |
0 |
0 |
3 |
|
Supraoccipital bone – hole |
19 |
19 |
20 |
28 |
|
Supraoccipital bone – b.o. |
0 |
0 |
0 |
14 |
|
Tympanic annulus – i.o. |
6 |
6 |
4 |
1 |
|
Tympanic annulus – short** |
0 |
0 |
0 |
3 |
|
Arcus zygomaticus – misshapen** |
0 |
1 |
0 |
0 |
|
Basisphenoid –i.o. |
0 |
0 |
1 |
0 |
|
Squamous part of temporal bone –i.o. |
0 |
0 |
0 |
1 |
|
STERNEBRA |
Sternebrae – decreased number of ossification sites* |
175 |
126 |
159 |
126 |
Sternebrae – ossification sites i.o. |
169 |
122 |
153 |
104 |
|
Sternebrae – ossification sites u. |
76 |
68 |
92 |
111 |
|
Sternebrae – ossification sites b.o. |
5 |
0 |
15 |
4 |
|
Sternebrae – ossification sites - asymmetric ossification |
0 |
4 |
2 |
2 |
|
Sternebrae – ossification sites - absent** |
0 |
1 |
0 |
0 |
|
Sternebrae – ossification sites – malpositioned** |
1 |
0 |
0 |
0 |
|
VERTEBRAE |
Vertebrae - thorac, lumb. centrum – d.o. |
2 |
3 |
5 |
4 |
Vertebrae - thoracic centrum – b.o. |
4 |
4 |
6 |
6 |
|
Vertebrae – thorac., lumb., sacr., - absent** |
0 |
2 |
1 |
0 |
|
RIBS |
Ribs – wavy |
5 |
5 |
2 |
0 |
Ribs – i.o. |
1 |
0 |
0 |
0 |
|
Ribs – supernumerary site |
17 |
17 |
11 |
7 |
|
Ribs – short |
0 |
1 |
0 |
0 |
|
Ribs – absent** |
0 |
3 |
3 |
3 |
|
Humerus – short** |
0 |
1 |
0 |
0 |
|
Scapula – i.o. |
8 |
4 |
0 |
0 |
|
Ilium – i.o. |
1 |
1 |
0 |
1 |
|
Pubis– i.o. |
0 |
1 |
0 |
5 |
|
Ischium – i.o. |
1 |
1 |
0 |
11 |
|
Note:Values statistically significant on probability level 0.05(p<0.05) are shaded.
* - the mentioned ossification sites are either incompletly ossified or unossified
** malformation
Effect on developmental toxicity: via oral route
- Endpoint conclusion:
- adverse effect observed
- Dose descriptor:
- LOAEL
- 125 mg/kg bw/day
- Study duration:
- subacute
- Species:
- rat
- Quality of whole database:
- The key study is an OECD/GLP guideline study in rats with a Klimisch score of 1. The quality of the database is high.
Effect on developmental toxicity: via inhalation route
- Endpoint conclusion:
- no study available
Effect on developmental toxicity: via dermal route
- Endpoint conclusion:
- no study available
Additional information
Pre-natal developmental toxicity
In a pre-natal developmental toxicity study (18-324), L-carvone (99.16%) was administered to 24 female Wistar CRL rats in corn oil by gavage at dose levels of 0, 125, 250 and 500 mg/kg b.w./day from days 5 through 19 of gestation.
There were no unscheduled mortalities during the study. A negative effect of test item treatment on the growth of maternal animals was observed at the highest dose 500 mg/kg b.w./day. The body weight was statistically significantly decreased in this group and this correlated with the statistically significantly decrease of food consumption. Adverse changes of health condition and clinical symptoms of intoxication were observed only in females at the dose 500 mg/kg b.w./day. Also, serious pathological findings and a statistically significant decrease in the absolute weight of the uterus were revealed at this dose level. These findings were clearly related to treatment by the test item. There were no changes in reproduction parameters in treated groups.
The examination of foetuses revealed a statistically significant decrease in the mean body weights at the dose 500 mg/kg b.w./day. The decreased weight of foetuses could be associated with a negative influence of the test item on the growth of maternal animals.
Examination of foetal skull and skeleton revealed anomalies in all treated groups. The incomplete ossification of the supraoccipital bone, could be associated with lower foetal body weight, especially at the highest dose level. Equally, transitional findings in the foetal skull, such as bipartite ossification of the supraoccipital bone and unossified supraoccipital bone could be associated with maternal toxicity, due to incidence at the dose 500 mg/kg b.w./day only. The increased incidence of transitional findings in all treated groups in comparison with control group – mainly hole in supraoccipital bone, asymmetric ossification of sternebra and bipartite and dumbbell ossification of vertebrae could be considered as adverse effect of the test item to foetuses, because no maternal toxicity was observed at the 125 and 250 mg/kg b.w./day dose levels. The malformations of ribs (absent) were observed only in treated groups.
The increased incidence of transitional findings (in foetuses of treated females compared to control females) could be considered as an adverse effect of the test item to early prenatal development of organism in uterus. The effect of the test item on the incidence of malformation of ribs in treated groups is disputable.
In a pre-natal developmental toxicity study in rats, the NOAEL for toxicity in pregnant females was established 250 mg/kg b.w./day. This NOAEL value is based on no occurrence of mortality, no changes in health condition status, no reproductive-related pathological findings and no changes in reproduction parameters at the dose 250 mg/kg b.w./day.
The NOAEL for prenatal development is < 125 mg/kg b.w./day. This NOAEL value is base on the occurrence of transitional findings of supraooccipital bone, sternabrae, vertebrae and malformations of the ribs at a dose of 125 mg/ kg b.w./day. The dose level 125 mg/kg b.w./day can be treated as the LOAEL in this study.
Justification for classification or non-classification
Considering be below reasons:
1) Transitional findings are not considered as adverse, and the incidences of these transitional findings at the low and medium dose groups are comparable to the control.
2) There could be concern regarding the absence of ribs found at all doses. However, due to the limitations of the study design (e.g. lack of detection of cartilage or later development of these foetuses), it is not completely clear if the absent ribs or vertebrae are a consequence of agenesis or simply delayed ossification.
The substance L-carvone (CAS No. 6485-40-1) is not classified for reproductive toxicity (Category 2) when the criteria outlined in Annex I of 1272/2008/EC are applied.
Additional information
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