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Administrative data

Description of key information

No reliable repeated study is available on TMTM. A combined 28 -day repeated toxicity study with reproduction screening study will be performed according to the ECHA decision.

By waiting for the results on TMTM, the read-across with Thiram is maintained to justify the classification and to derive DNELs.

Key value for chemical safety assessment

Toxic effect type:
dose-dependent

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
Group of 64 males and 64 female rats were given thiram at constant dietary doses of 3, 30 and 300 ppm (0, 0.1, 1.2 and 11.6 mg/kg/d for males, and 0, 0.1, 1.4 and 13.8 mg/kg/d for females) for 104 weeks.
GLP compliance:
no
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Japan CLEA Co. (Tokyo)
- Age at study initiation: 5 weeks of age
- Weight at study initiation: no data
- Housing: by four in stainless-steel cages with wire-mesh floors
- Diet (e.g. ad libitum): free access to the test diets
- Water (e.g. ad libitum): free access to tap water
- Acclimation period: 1 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C):23+/-1°C
- Humidity (%):55+/-10%
- Air changes (per hr):12
- Photoperiod (hrs dark / hrs light): lighting from 5:00 Am to 7:00 pm
Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
Since the compound was stable in test diets for 1 week, test diets were preapred twice a week during the treatment.
The concentrations of the compound in test diets were not adjusted to the growthcurve of the animals during the treatment.
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
104 weeks
Frequency of treatment:
daily (in diet)
Remarks:
Doses / Concentrations:
3, 30 and 300 ppm (0, 0.1, 1.2 and 11.6 mg/kg/d for males, and 0, 0.1, 1.4 and 13.8 mg/kg/d for females)
Basis:
nominal in diet
No. of animals per sex per dose:
64 rats/ sex/dose
Control animals:
yes, sham-exposed
Details on study design:
- Dose selection rationale: The dose levels were set on the basis of the results of a 4-week preliminary study in which groups of five male and five female rats were given test diets of 0, 10, 100,1000 and 2500 ppm. Males in the 100 ppm group and males and females in the 1000 and 2500 ppm groups showed moderate to marked unpalatability of the test diets with was reflected by retarded growth.
- After week 13, 26 and 52, eight males and eight females in each group were randomly selected and killed for the laboratory examinations.
Positive control:
no
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: yes, no details.

DETAILED CLINICAL OBSERVATIONS: yes, no details.

BODY WEIGHT: Yes, weekly up to week 26, biweekly up to week 52, and every 4 weeks thereafter.

FOOD CONSUMPTION : Yes, weekly up to week 26, biweekly up to week 52, and every 4 weeks thereafter.

WATER CONSUMPTION : Yes, weekly up to week 26, biweekly up to week 52, and every 4 weeks thereafter.

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: scheduled to be killed
- Dose groups that were examined: all rats

HAEMATOLOGY: Yes
- Time schedule for collection of blood: scheduled to be killed
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: all rats
- Parameters checked : hematrocrit, hemoglobin, erythrocyte count, leukocyte count, and differential leukocyte count.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: scheduled to be killed
- Animals fasted: Yes No data
- How many animals: all rats
- Parameters checked: total protein, alkaline phosphatase, glucose, blood urea nitrogen, glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, total cholesterol and calcium.

URINALYSIS: Yes
- Time schedule for collection of urine: scheduled to be killed
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- How many animals: all rats
- Parameters checked: pH, protein, glucose, ketones, and occult blood.

NEUROBEHAVIOURAL EXAMINATION: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
ORGAN WEIGHT : brain, pituitary, thyroid, heart, thymus, liver, kidneys, spleen, adrenals, testes, ovaries and calf muscle
HISTOPATHOLOGY: Yes
Other examinations:
no
Statistics:
Mortality was evaluated by a life-table analysis. Body weight, food intake, water intake, hematology, blood biochemistry, and organ weight were analyzed by the Student t test. The Mann-Whitney U test was used for urinalysis data. Ophtalmology and pathological findings were assessed by the Fisher exact probability test. A 5% level of probability was used as the criterion of significance.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
no clinical signs, but mortalities
Mortality:
mortality observed, treatment-related
Description (incidence):
no clinical signs, but mortalities
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Food efficiency:
no effects observed
Water consumption and compound intake (if drinking water study):
no effects observed
Ophthalmological findings:
no effects observed
Haematological findings:
effects observed, treatment-related
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
effects observed, treatment-related
Details on results:
CLINICAL SIGNS AND MORTALITY
Clinical signs indicative of toxicity were not observed in the rats of either sex during the treatment.
The mortality of females in the 30 and 300 ppm groups was slightly higher than that of the control group during the last 8 weeks of treatment. These higher mortality rates were apparently correlated to incidental higher occurrences of pituitary tumors during this period, although overall incidences of the tumors in these females were comparable to that of the control group (Tables 5 and 6).

BODY WEIGHT AND WEIGHT GAIN
In the 300 ppm group, the body weights of both sexes were remarkably depressed to rates of 10% or more of the controls during the early weeks of the treatment and remained at significantly lower levels during almost all the treatment.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
Food intake of the 300 ppm group was 30% or more lass than that of the controls for both sexes at the first week of treatment (table 1).
Although the animals began to recover their appetite, average daily food intakes in the 300 ppm group were 8.4 and 11.0% lower than those of the controls for males and females, respectively.
Chemical intake per body weight was significantly decreased during the first 13 weeks if treatment due to te rapid weight gain of the animals (Table 1), and the average values in the 3, 30 and 300 ppm groups during the entire treatment period were 0.1, 1.2 and 11.6 mg/kg/day for males and 0.1, 1.4 and 13.8 mg/kg/d for females.

WATER CONSUMPTION
No treatment-related changes in water intake (table 1) was observed.

OPHTHALMOSCOPIC EXAMINATION
No treatment-related changes in ophtalmology was observed.

HAEMATOLOGY
Upon hematologic examination, females of the 300 ppm group revealed decreases or decreasing tendencies in hematocrit, hemoglobin and erythrocyte count at interim kills after weeks 13, 26 and 52, although the values of these parameters were comparable to those of the controls at terminal kill (table 2).
Hematology examination of males in this group revealed no significant changes except for a slight decrease in erythrocyte count at interim kill after week 13 (table 2).

CLINICAL CHEMISTRY
Blood biochemistry examinations revealed incidental increases in GPT for both sexes in all treated groups at terminal kill, although the values were comparable to those of the controls in the examinations at three interim kills during the treatment (Table 2).
Incidental increases in GOT were observed in the 30 ppm group females and in the 300 ppm group males only at terminal kill.
There were no abnormal changes in other parameters, including ALP and BUN for all treated groups of both sexes during the treatment.

URINALYSIS
There were no changes in urinalysis attributable to the treatment.

ORGAN WEIGHTS
In organ weight analyses, a significant change in the calf muscle was observed (Table 4). In males, the calf muscle seemed to develop in proportion to growth of the body during the first year of life and the relative weights to body weights were almost the same among the three interim kills, weeks 13, 26 and 52, whereas the absolute weights increased with age during this period. Both relative and absolute weights of the tissues decreased nearly 40% during the next year, although body weight was almost stable during this period. Relative weight showed a tendency to decrease with prolonged treatment. The values decreased in the latter half of the treatment, showing 7 and 12% decreases for absolute and relative weight, respectively. In the 300 ppm group, both absolute and relative weights of the calf muscle were significantly decreased in both males and females at scheduled kills after weeks 52 and 104, although they were comparable to the controls at interim kills after week 13 or 26.
Kidney weight of males and females showed a tendency to decrease in the latter half of the treatment (Table 4).
Females in this group exhibited an increased thryoid weight at terminal kill. The thyroid weights of females in the 30 and 3 ppm groups appeared to be increased at terminal kill, corresponding to the greater body weight of these animals.
No treatment-related changes in the weights of the liver, testes, or ovaries were observed.

GROSS PATHOLOGY
At necropsy, the lowered weights of the calf muscles appeared to correspond to an increased incidence of atrophy of the calf muscles in females of 300 ppm group, although males did not show an increased occurrence of this change (Table 5).
A striking change in gross lesions observed at terminal kill was that only 1 of 20 females in the 300 ppm group had skin mass versus 16 of 28 females in the control groups (Table 5). The overall incidence of skin mass was significantly decreased in females of the 300 ppm group, 7 of 64, compared with the control group, 19 of 64. In females of 30 ppm group, occurrence of skin mass was also considerably depressed, 7/20 versus 16/28 in the control group, at terminal kill, although there was no statistically significant difference between the groups. In females these skin masses arose largely on the ventral or dorsal surface of the body, with considerably higher occurrences at the axillary, lower abdominal, or upper thigh region. They appeared to be solid nodular or multilobular and sometimes had cysts of various sizes containing brownish-white, proteinous, or milky materials.

HISTOPATHOLOGY: NON-NEOPLASTIC
In histology, atrophy of the calf muscle observed at necropsy was revealed to be atrophy or degenerative changes of muscle fibers of the tissue (Table 5).The changes were frequently observed in females found dead or killed in extremis in the last quarter of the treatment as well as in females at terminal kill. The muscle fibers were atrophic, losing their acidophilic tincture and were replaced partially by collagen fibers. The lesions were limited in the calf muscle but not in the muscles of other parts of the body, including the thigh. A variety of degenerative processes including atrophy, vacuolation, fibrosis, and mineralization of the nerve fibers were found in the sciatic nerve in the aged animals of both treated and control groups. In the last quarter of the treatment period, the lesions were observed more frequently in the females of the high-dose group than in the control group. In most cases the muscular lesions of the calf muscle were found concomitant with the alterations in the sciatic nerve. there were no abnormalities in the histology of the osseous and cartilaginous tissues from the sternum and femur.
In males, the high-dose group showed a significantly depressed occurrence of myocardial atrophy/fibrosis, which is a common senile change of male rats of this strain (table 5). In females, the incidence of this lesion was incidentally increased in the 3 or 300 ppm groups. occurrence of chronic nephrosis, which is also a typical senile lesion of the rat, was considerably depressed in females of the 300 ppm group (Table 5), but no similar tendency was observed in males. There were no remarkable changes in the pancreas, thyroid and testis.
During the last 8 weeks of the treatment period, pituitary adenoma occurred more frequently in females of the 30 and 300 ppm groups (4 of 5 and 7 of 8, dead animals bore the tumor respectively) than in those of the 3 ppm and control groups (one of 3 in both groups), resulting in slightly higher mortalities in middle- and high-dose females during this period. However, since the occurrence of this tumor in these two female groups at terminal kill was less than that in the 3 ppm and control groups, overall incidences of the tumor were comparable in all groups (table 5).
The decreased occurrences of skin masses in females of the middle- and high-dose groups were comparable to the decreased incidences of mammary fibroadenoma (tables 5 and 6). Although this negative trend in incidence of this tumor was not confirmed statistically for overall incidence of females in the 30 ppm group, the incidence (9/64) was lower than that of the control (16/64). the incidence of mammary tumors was also considerably depressed in females of the 3 ppm group at terminal kill, 9/29 versus 15/28 in the control group, although no statistical significance was seen. The incidences of other types of tumors were comparable to those of the controls in both sexes (table 6).
Dose descriptor:
NOEL
Remarks:
systemic effect
Effect level:
0.1 mg/kg bw/day (actual dose received)
Sex:
male/female
Basis for effect level:
other: 3 ppm
Dose descriptor:
NOAEL
Effect level:
1.2 mg/kg bw/day (actual dose received)
Sex:
male/female
Basis for effect level:
other: = 30 ppm Based on adverse effects observed at 11.6 mg/kg/day (300 ppm).
Critical effects observed:
not specified
Conclusions:
The NOAEL for long-term toxicity was 1.2 or 1.4 mg/kg bw/day for males or females, respectively
Executive summary:

In 2-year rat study, Wistar rats (n=64/sex/group) received dietary doses (0, 3, 30 and 300 ppm) of thiram (purity: 98.7%) equivalent to 0, 0.1, 1.2, or 11.6 mg/kg bw for males and 0, 0.1, 1.4, or 13.8 mg/kg bw for females. The mortality of females was dose-dependently increased in the 13.8- and 1.4-mg/kg bw groups during the last 8 weeks of the treatment period. Body weight gain and food consumption were decreased in both males and females of the high-dose group. During the last part of the test period, food consumption was also decreased in males receiving 1.2 mg thiram/kg bw. In females of the high-dose group, haemoglobin and haematocrit levels and red blood cell count were significantly decreased at week 26, but no differences were found at termination (week 104). At week 104, slightly but significantly increased plasma ALAT levels were found in all treated groups (both sexes) and a slightly, significantly increased plasma ASAT level in high-dose males and in mid-dose females. No changes were observed in plasma alkaline phosphatase and urea levels. The committee considered the liver enzyme changes to be incidental to treatment. At termination, absolute kidney weight, and absolute and relative calf muscle (M. triceps surae) weights were decreased in high-dose males. Females showed significantly increased absolute thyroid weights in all treated groups, but significantly decreased absolute liver, kidney, and calf muscle weights at the high dose only. Microscopic examination revealed a significant increase in atrophy and degeneration of this muscle in females of the high-dose group. This effect was considered to be secondary to the atrophy and degeneration of the sciatic nerve that was also observed in high-dosed females (n=15). Pituitary adenomas were observed in 4/5 mid-dose and 7/8 high-dose females, which were found dead during the last 8 weeks of the treatment. Since the incidence of these tumours in these 2 female groups at terminal kill was less than in the low-dose or control groups, overall incidences of the tumour were comparable in all groups. In addition, this type of tumour is often seen in ageing rats, and, therefore, the committee did not consider these tumours to be related to thiram treatment. The incidence of fibroadenomas of the mammary gland in females was significantly decreased in a dose-related fashion in the high-dose group. The incidences of other tumours were comparable to those of controls in both sexes. No evidence for carcinogenic effects of thiram was found. The NOAEL for long-term toxicity was 1.2 or 1.4 mg/kg bw/day for males or females, respectively

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
1.2 mg/kg bw/day
Study duration:
chronic
Species:
rat
Quality of whole database:
Maita's study is a reilable study with a klimisch score of 2.
Organ:
liver

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Chronic study on thiram

In the key study, Wistar rats (n=64/sex/group) received dietary doses (0, 3, 30 and 300 ppm) of thiram (purity: 98.7%) equivalent to 0, 0.1, 1.2, or 11.6 mg/kg bw for males and 0, 0.1, 1.4, or 13.8 mg/kg bw for females for 2 -year. The mortality of females was dose-dependently increased in the 13.8- and 1.4-mg/kg bw groups during the last 8 weeks of the treatment period. Body weight gain and food consumption were decreased in both males and females of the high-dose group. During the last part of the test period, food consumption was also decreased in males receiving 1.2 mg thiram/kg bw. In females of the high-dose group, haemoglobin and haematocrit levels and red blood cell count were significantly decreased at week 26, but no differences were found at termination (week 104). At week 104, slightly but significantly increased plasma ALAT levels were found in all treated groups (both sexes) and a slightly, significantly increased plasma ASAT level in high-dose males and in mid-dose females. No changes were observed in plasma alkaline phoshatase and urea levels. The committee considered the liver enzyme changes to be incidental to treatment. At termination, absolute kidney weight, and absolute and relative calf muscle (M. triceps surae) weights were decreased in high-dose males. Females showed significantly increased absolute thyroid weights in all treated groups, but significantly decreased absolute liver, kidney, and calf muscle weights at the high dose only. Microscopic examination revealed a significant increase in atrophy and degeneration of this muscle in females of the high-dose group. This effect was considered to be secondary to the atrophy and degeneration of the sciatic nerve that was also observed in high-dosed females (n=15). Pituitary adenomas were observed in 4/5 mid-dose and 7/8 high-dose females, which were found dead during the last 8 weeks of the treatment. Since the incidence of these tumours in these 2 female groups at terminal kill was less than in the low-dose or control groups, overall incidences of the tumour were comparable in all groups. In addition, this type of tumour is often seen in ageing rats, and, therefore, the committee did not consider these tumours to be related to thiram treatment. The incidence of fibroadenomas of the mammary gland in females was significantly decreased in a dose-related fashion in the high-dose group. The incidences of other tumours were comparable to those of controls in both sexes. No evidence for carcinogenic effects of thiram was found. The NOAEL for long-term toxicity was 1.2 or 1.4 mg/kg bw/day for males or females, respectively.


Justification for classification or non-classification

TMTM is not classified in the Annexe VI of CLP for the repeated toxicity.

But TMTD is classified in the Annexe VI of CLP as STOT RE 2 with "liver" as target organ by oral route.

Since a read-across is proposed for the repeated toxicity endpoint, TMTM must be classified like as TMTD.

Regulation (EC) No 1272/2008: STOT RE 2, H 373 (May cause damage to organs through prolonged or repeated exposure).

Target organ = Liver