Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Weight of evidence: According the National Institute of Technology and Evaluation for the test substance tested in concentration 0. 0.4, 2 and 10 mg/kg the NOAEL and NOEL was assigned the value of 2 mg/kg since no changes attributed to the test substance were noted in the parental animals and test substance does not cause any abnormalities on the offspring. For the test substance tested in the concentration 0,0.2, 1 and 5 mg/kg, the NOAEL and NOEL value were judged to be 1 mg/kg since nursing failure of the dams, and increase in stillbirth and a decrease in viability indices on days 0 and 4 after birth were observed in the group of 5 mg/kg.No abnormalities were detected in parental animals nor in the offsprings in tested concentrations of Thiosemicarbazide.

Link to relevant study records

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Endpoint:
one-generation reproductive toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
September 25, 2007 - February 20, 2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP, Guideline followed
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Test material information:
Composition 1
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Refrigerated:3.5-5.9ºC, range 1-10ºC, dark place
- Stability under test conditions: Yes
Species:
rat
Strain:
Crj: CD(SD)
Details on species / strain selection:
Reason for selecting lineage, It is widely used for toxicity tests using rodents, has abundant background data.
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:Charles River Japan Co., Ltd. (Atsugi Production Center)
- Age at study initiation: 9 weeks old
- Weight at study initiation:Male:315-366 g, Female: 205-234g range of the animals was within ± 20% of the average body weight.
- Fasting period before study: Yes, the day before the planning dissection was fasted (about 18 to 23 hours).
- Housing: Cages, equipment made by Tokiwa Scientific Instruments Co., Ltd. was sterilized by autoclaving used. However, steel stand is excluded.Using a stainless steel hanging metal gauze cage (195 W × 325 D × 180 H mm). During the pregnancy - using a polycarbonate cage (265 W × 426 D × 200 H mm),
- Diet (e.g. ad libitum): Radio-sterilized solid feed for laboratory animals (CRF-1, Oriental Yeast Co., Ltd.. ). Free intake except for spontaneous momentum measurement and fresh urine collection and exchange at feeder replacement.
- Water (e.g. ad libitum): Drinking water,After filtering by 5 μm filter, ultraviolet irradiated tap water.Free intake was excluded except for spontaneous momentum measurement and fresh urine collection. Drinking water in the water bottle. During the pregancy:Polycarbonate water bottle (700 mL) was used. Water supply device Tokiwa Scientific Instruments Co., Ltd.
- Acclimation period:The animals were in quarantine/acclimation period for 5 days, observed every day for any abnormalities.

DETAILS OF FOOD AND WATER QUALITY:
Water Quality: Mitsubishi Chemical Analytech Co., Ltd. (former company name: Diamond Analysis Center Co., Ltd.) The inspection is carried out periodically (twice a year), and the obtained analysis value becomes the standard operation manual.
Food Quality: Analysis results conducted at the Japan Food Research Laboratories Foundation, analyzed using Oriental Yeast Co., Ltd. The concentrations of pollutants such as residual agricultural chemicals in lots used were obtained from the company,It meets the standards.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):21.0-23.2ºC (tolerance range 19.0 to 25.0 ºC)
- Humidity (%):47.8-61.3% (tolerance range 35.0 to 75.0%)
- Air changes (per hr):6 to 20 times / hour, all fresh air supply
- Photoperiod (hrs dark / hrs light):12 hours / day (7: 00-19: 00)
Route of administration:
oral: gavage
Vehicle:
water
Remarks:
Purified
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:Dosing liquid volume: Ten mL / kg, and the liquid volume of each individual was calculated based on the body weight measured on the nearest day.Preparation was carried out in a dispensing room under illumination with ultraviolet rays cut off. Frequency of preparation is test facility. Based on the results of the stability analysis carried out in 7 days, test was conducted once.
Details on mating procedure:
- M/F ratio per cage:1:1 for each group
- Length of cohabitation: 8 days
- Proof of pregnancy:sperm in vaginal smear referred to as day 0 of pregnancy.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analytical method validation of Hydrazinecarbothioamide in preparation liquid:
After preparation, dispense into polypropylene tubes at each administration day and limit the period of time when stability was confirmed.
Refrigerated (measured value: 3.5 to 5.8 ° C, tolerance: 1 to 10 ° C) • Storage in a dark place
1 mg / mL preparation solution:
A predetermined weight of the test substance was weighed, an appropriate amount of the medium was added, stirred, and dissolved. Thereafter, 1 mg / mL was added using a metric glass or a measuring cylinder to make the concentration of female.
0.2 and 0.04 mg / mL preparation:
1 mg / mL solution was prepared by diluting with a medium.

Confirmation of stability
The stability of the test substance in the administration solution under refrigeration, in the dark, and under stoppered condition is 0.01 and 5 mg / mL conducted at the test facility for 8 days.

Confirmation of Concentration
The administration solution at the time of initial preparation was analyzed by Spectrophotometer: Model HITACHI U-3310, Cell: 10 mm quartz cell (n = 2), and the average value of the concentration (actually measured value: 99.4 ~ 106.0%, setting concentration within ± 10%)

Standard and sample solution were prepared.

Measurement of standard solution
Absorbance of standard solutions (ST-1, ST-2 and ST-3) at 235 nm was measured by spectrophotometry with water as a control and a calibration curve was prepared from the concentration of the standard solution.

Measurement of sample solution
The absorbance of the sample solution at 235 nm was measured by spectrophotometry using water as a control, Calculate the HCTA concentration in the sample solution from the calibration curve and the dilution factor, calculate the average value.

Recovery period
In the control group and 5 males and 5 females in the 10 mg / kg group, a recovery period of 14 days after the administration period was administered. However, females were set as satellites without mating.
Duration of treatment / exposure:
MALES: 42 days
FEMALES: 42 - 50 days (from 14 days before mating to day 4 of lactation)
FEMALES (satellite) : 42 days
Frequency of treatment:
Daily
Details on study schedule:
- F1 parental animals not mated until 15th days.From the evening on the 15th day to the longest 8 days were mated.
Dose / conc.:
0 mg/kg bw/day (nominal)
Dose / conc.:
0.4 mg/kg bw/day (nominal)
Dose / conc.:
2 mg/kg bw/day (nominal)
Dose / conc.:
10 mg/kg bw/day (nominal)
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
RECOVERY
Dose / conc.:
10 mg/kg bw/day (nominal)
Remarks:
RECOVERY
No. of animals per sex per dose:
MALES: 12 (5 for recovery) in each group
FEMALES: 12 in each group
SATELLITE FEMALES: 5 for each group
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Based on the results of range finding study.

Three doses of 0.4, 2 and 10 mg / kg. It was set with reference to the result of the dose setting test conducted at the test facility. "14-day repeated dose toxicity by oral administration of Hydrazinecarbothioamide in rats Test (test number: B070687) "
FRIST TRIAL: Dose: 0, 0.4, 2 and 10 mg / kg,
Number of animals: each group: 1female, Three male,
The low value of the thymus weight and the low value of the body weight are not significant although there is no significant difference in the males at 10 mg / kg. Slightly high heart weight was found in females of 10 mg / kg group. Because clear expression of toxicity could not be confirmed in this study, 50 mg / kg was used for the highest dose.

Dose setting test (test number: B070903) was performed again. "14-day repeated dose toxicity by oral administration of Hydrazinecarbothioamide in rats
Test (test number: B070903) "
SECOND TRIAL: Dose: 0, 20, 30 and 50 mg / kg,
Number of animals: each group: 3 males and 3 females
All sexes of 20 mg / kg or more died on the 1st day or moribund. It was a state. These animals showed irritability and tonic convulsions, the breathing irregularity. As a result of necropsy, pulmonary edema of 20 mg / kg or more. Pancreatic fluid reserves in males and females in the 20 and 30 mg / kg group and males in the 50 mg / kg group was observed. As a result, although death was observed in the 20 mg / kg group, death occurred in the 10 mg / kg group, No moribund was observed, and changes that could be attributed to the test substance could be confirmed.
Therefore, 10 mg / kg at which some toxicity is expected to occur is set to a high dose, and three doses were set for the lower common ratio 5, medium dose 2 mg / kg, low dose 0.4 mg / kg. Also, a control group to which only body (purified water) was administered was provided.
First trial was selected as a tested doses.
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes, in the males and females (Tremor, clonic convulsion, breathing)

DETAILED CLINICAL OBSERVATIONS: Yes, observation of reacting from removal from the cage, reactivity to handling, aggression, trauma, color of skin, soiled fur, exophthalmos, palpebral closure, color of conjumctuiva, secretion,lacrimation, salivation, piloerection, pupil size, arousal, urination, defecation, posture, body position, gait, bizzarre behaviour, tremor, clonic convulsion,tonic convulsion, breathing, stereotypy.
- Time schedule: 9 weeks (from week -1 to week 8)

BODY WEIGHT: Yes,
Males animals were treated on days 1, 8, 15, 22, 29, 36, 42 and 43, In addition, in male-recovered animals it was measured on days 50 and 56. Female satellite animals (the same as males) are recovered from male,measured at the same frequency. Female test animals - 0, 7 and 14 days before mating, during gestation - 0, 7, 14 and 20 days, lactation - 0 and 4 days.
Measurement - an electronic balance weight (EB - 3200S: Shimadzu Corporation, PB 3002 - S: Metlitore Co., Ltd.) was used.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
Males animals were treated on days 8, 15, 29, 36, 40. In addition, in male-recovered animals it was measured on days 50 and 54.
Female satellite animals were treated on days 8, 15, 22, 29, 36, 42, recovered 50 and 56 days.
Female test animals - 7 and 14 days before mating, during gestation - 7, 14, 20 days, and during lactation - 4 days.
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Not specified
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes
Measurement - an electronic balance weight (EB-3200S: Shimadzu Corporation, PB 3002-S : METTLER TOLEDO CO., LTD.)

OTHER:

HAEMATOLOGY: Yes (Red/White blood cell count, hemoglobin concentration, hematocrit, MCV, MCH, MCHC, platelet count, reticulocyte ratio, PT, APTT, Lymphocute, neutrophil, eosinphil, basophil, monocyte )
Males, Females:day 43 and 57

CLINICAL CHEMISTRY: Yes (ASAT(GOT), ALAT(GPT), GAMA-GT, ALP, Total bilirubin, urea nitrigen, creatinine, glucose, total cholesterol, tryglyceride, total protein, albumin, A/G ratio, calcium, Inorganic phosphorus, Na, K, Cl)
Males, Females:day 43 and 57

URINALYSIS: Yes, only in males on day 40: (pH, protein, glucose, ketones, bilirubin, occult blood, urobirinogen)

NEUROBEHAVIOURAL EXAMINATION: Yes,the day before the administration, once a week until the 6 th week both after 13: 00
Touch response, auditory respione, tail pinch response, aerial righting reaction, forelimb, hindlimb
Estrous cyclicity (parental animals):
Yes, observed for regularity and irregularity in every concentrations in males and females before mating and during mating period. (Implantations, gestation length and parturition)
Sperm parameters (parental animals):
No specified.
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups,stillbirths, live births,viability index on day 4, body weights

GROSS EXAMINATION OF DEAD PUPS:
yes, for external abnormalities; possible cause of death was determined for pups born or found dead.
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: day 43 of treatment and day 15 of recovery
- Maternal animals: day 5 of lactation, Females (satellite), day 15 of recovery

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations.(Absolute and relative weight of liver, kidney, thymus)

HISTOPATHOLOGY / ORGAN WEIGHTS
The following organs and tissues of all animals were collected and fixed with 10 vol% neutral phosphate buffered formalin solution.The testes and epididymis of male were fixed with Bouin's solution, then 10 vol% and stored in neutral phosphate buffered formalin solution.
Brain, pituitary gland, thymus, lymph node (mandible / mesenteric), trachea, lung, stomach, intestinal tract (twelve fingers Intestine, jejunum, ileum, cecum, colon, rectum), thyroid / parathyroid (bilateral), heart, liver, Spleen, kidney (both sides), adrenal gland (both sides), bladder, testis (both sides), epididymis (both sides), semen Bulla (including coagulated gland), prostate ventral lobe, ovary (both sides), uterus, vagina, bone marrow (right side femur) Sciatic nerve (right side), spinal cord, grossly abnormal site

Males:Day 43 and 57,Organ weight and relative organ weight :(Final body weight, brain, thymus, heart, liver, spleen, kidneys, adrenals, testes, epicidymides)
Satelite females: on day 57, Organ weight and relative organ weigh(Final body weight, brain, thymus, heart, liver, spleen, kidneys, adrenals)
Postmortem examinations (offspring):
SACRIFICE
- On day 4 after birth

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations. Necropsy was performed on dead children, immersed in 10 vol% neutral phosphate buffered formalin solution.Children who died unable to withstand the examination by killing were excluded.

Statistics:
Data on newborn babies was calculated for each mother animal as a sample unit.
Weighed data was analyzed - Bartlett Method. If the variances are equal, one-way analysis of variance, dispersion is performed. If not equal, Kruskal-Wallis's test is used. If there is a significant difference between the groups,Dunnett method or Dunnett type multiple comparison is used. For multi-group counting data in urinalysis, Kruskal-Wallis's test. When significant difference-Dunnett type multiple comparison The counting data in the academic examination were compared between the two groups with the control group in Wilcoxon rank sum test. Other counting data were tested by Fisher's direct stochastic method. Data of two groups of recovered animals (control group and 10 mg / kg group) were analyzed as follows. The data is tested for equal variance by F test, and if the variances are equal, Student's test. When the test and the variance are not equal, Aspin-Welch's test used. The significance level of each test was 5%. The test is carried out using a safety test system (MiTOX, MES System Engineering Co., Ltd.). The target of statistical analysis: general condition, behavioral inspection (details Responsiveness to stimulation of fine symptoms observation, function test) and autopsy findings. Multiple comparison test: Body weight, food intake, hematology examination, blood biochemical examination, organ weight, behavior inspection measurement data (gras power, locomotor activity), number of corpus luteum, number of landing, number of births (number of babies born, number of stillborn infants). Multiple comparison of Kruskal-Wallis and Dunnett type: Urinalysis.
Fisher's direct stochastic method: Expression rate of abnormal periodic animal, mating, conception, birth, sex ratio (male/(male+female), group total number), the incidence of maternal animals with abnormal children. F test and Student or Aspin-Welch's t test: Weighing data of recovered animals
Reproductive indices:
Copulation, fertility,delivery, implantation and gestation index were examined.
Offspring viability indices:
Viability indices on day 4 were observed. Survival, litter loss, sex ratio and number of stillborns were observed.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
No clinical abnormalities throughout the administration and recovery periods, tonic convulsion was observed in 2 males and 1 female in the 10 mg/kg group at detailed clinical observation.
One male in the 10 mg/kg group showed a hypersensitive reaction in touch response and tail pinch response
Observation within home cage, observation at handling and within open field: No abnormalities were found in any animals at the time of observation.

Dermal irritation (if dermal study):
not examined
Mortality:
mortality observed, treatment-related
Description (incidence):
One female in the group of 10 mg/kg/bw was found dead before dosing on day 2 of administration. She had pulmonary edema.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
The body weight gain was comparable to that of the control group from day 15 onwards. It only occurred in the early stage of administration and recovered thereafter. In the 0.4 and 2 mg / kg groups, there was no significant difference between the sexes and the control group.

Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Throughout administration and recovery period in both males and females, there was a significant difference between the control group and the test substance administered group.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
No change due to the test substance was observed.
At the end of administration period
A low tendency of white blood cell count was observed in males of 10 mg / kg group,as a result, all cases were within the range of the average value of the background data ± 2 S. D. (accidental change)
In addition, the low value of MCHC and shortening tendency of PT were observed in males of the 10 mg / kg group. Only for MCHC, it is slight change compared with the control group, and hemoglobin concentration and the hematocrit value was almost the same as that of the control group, and for PT, the control group one case of high value, it is a change opposite to extension which is usually toxicologically problematic. As a result, it was judged that any changes were not toxicologically meaningful. Also, in the 0.4 mg / kg group extension of PT in females, prolongation of APTT in females at 0.4 and 2 mg / kg group, white in females at 2 mg / kg group. A low value of blood cell count was observed. However, neither correlation with the dose is recognized. (accidental change)

At the end of the recovery period
There was no significant difference between the test items in both sexes and the control group.

Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
At the end of administration period
High levels of total cholesterol were observed in males in the 10 mg / kg group. In addition, low or low trends of ASAT and ALAT were observed in males of the 10 mg / kg group. Since it is the opposite change from the high value which is usually toxicologically problematic, it is taken as a emergent change. Also, ALP and creatinine were elevated in males of the 0.4 mg / kg group. Although it was not observed correlation with the dose, it was determined to be an accidental change.

At the end of the recovery period
A low value of total protein and a high value of inorganic phosphorus were observed in males of the 10 mg / kg group. No change is recognized at the end, background data of the test facility. As a result of comparison with the total protein, all cases were within the average value of background data ± 2 S. D. Inorganic phosphorus showed a high value in 1 case, but in other test items since no relevant changes are noticed, it is judged that there is no toxicological significance
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
In the urine qualitative test, in any of the test items, between the control group and the test substance administered group, there was no significant difference between them.
Behaviour (functional findings):
effects observed, treatment-related
Description (incidence and severity):
One male in the 10 mg / kg group, at the time of examination of contact reaction and tail pinch reaction behind the act of crying was observed. Tactile reaction: Violent reaction, tail pinch reaction: jump, reaction to exaggeration in the 0.4 and 2 mg / kg groups, in both sexes. In the grip force measurement, no significant difference was observed between the control group and the test substance-administered group. In the examination during the recovery period, these changes were not observed and abnormalities were found in any animals

In the 10 mg / kg group two males and one female showed tonic convulsions in the 6th week.It was observed at the time of maintenance, at the time of function test execution or after returning to the cage after the examination was ended. In the 0.4 and 2 mg / kg group, no abnormality was observed in any animals in both males and females. These changes were not observed in the examination during the recovery period, and abnormalities were observed in any animals.

Measurement of locomotor activity
There was no significant difference between the control group and the test substance administration group in both males and females.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
At the end of administration period
Absolute and relative weight, low and low thymus in males in 2 and 10 mg / kg group. Value trend or high trend of liver and kidney was observed in males of 10 mg / kg group.
At the end of the recovery period
In the males of the 10 mg / kg group, the relative weight of the brain was high. The absolute weight is a similar value and a low value of the final body weight is seen, it is attributed to a low value of body weight.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
No change due to the test substance was observed. Testicular and epididymal compacts were bilateral in one case of 2 mg / kg group, one in 10 mg / kg group It is taken as change not related to the test substance.
Besides, dark red spots or red spots of the lungs are administered or after the recovery period finishes in the animal, 2, 1, and 1 males in the 0.4, 2 and 10 mg / kg group There was a deformity of the unilateral adrenal glands in the group of 0.4 mg / kg. It was found in one female group. All of these changes are sporadic emergence without correlation with dose. Since it is present, it is judged that there is no toxicological significance.

In the death animal:Pulmonary edema accompanied by increase of pleural effusion was observed as a change caused by the test substance. In addition, hepatic diaphragmatic surface nodules were observed, but this is a spontaneous change and contingent.
In the non-delivering animal: No change.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
One female in the group of dosing 10 mg/kg had pulmonary edema (dead)
The pulmonary edema was seen in all dead or moribund animals on day 1 of administration at 20 mg/kg and above in a preliminary study.

Planned anatomy
Changes due to the test substance were observed in the male thymus group of 10 mg / kg group. Accepted change.
In males in the 10 mg / kg group, atrophy of the thymus was observed in 3 cases. After the recovery period the dissected animals, this change was not recognized.
In the female spleen of the 10 mg / kg group, 3 cases showed a slight erythroidic extramedullary hematopoiesis enhanced. This change was seen in 1, 3, 1 cases in the control group, 0.4 and 2 mg / kg group, respectively. It was not correlated with the apparent frequency and frequency of expression. In addition,
Increased hematopoiesis is also known to develop with pregnancy. Therefore, it was determined not to be caused by the pregnancy, but to be expressed with pregnancy. In addition, 0.4 mg / kg in one female group, focal fibrosis was observed in the unilateral adrenal glands which had deformed at autopsy.
Dark red spots of the lung were seen in 2, 1, 1 cases of males in the 0.4, 2 and 10 mg / kg groups, respectively. The red plaques are localized bleeding, and from the number of examples of expression it is related to the test substance. It is admited as no change. Also, in the testes, the diffuse atrophy of seminiferous tubules was 2 mg / kg group, one case (mild, bilateral), 1 case of 10 mg / kg group (moderate, one side). Localized degeneration (minor) of seminiferous tubules was observed in the remaining unilateral testis of the group. However, there was no change in the testes in the other 10 mg / kg group, and these atrophy / degenerative findings were raren the testis but also in the background, it is judged that it is not related to the test substance.
In addition, various tissue changes were observed. However, they occur nonspecifically in rats, it is a present change, and it is not correlated with an obvious dose in the number of cases of expression, it was judged that the change was not related to the test substance.

Dead Animal: Moderate pulmonary edema was observed as a possible change in the cause of death. In addition, spleen white- A slight increase in the nuclear fracture image was observed in the splenic cord, mandibular lymph node and mesenteric lymph node. At the necropsy, hepatic diaphragmatic nodules found in the liver showed minor fibrosis at the base.
Histopathological findings: neoplastic:
not specified
Other effects:
not examined
Reproductive function: estrous cycle:
no effects observed
Description (incidence and severity):
In the sex cycle examination, there is no change in the mean periodic days, and the sex cycle is extended by the test substance.
No shortening was observed. There was no significant difference between the control group and the test substance administered group in any of the following rates: The copulation rate, the number of days required for mating, the number of estrus periods missing the copulation.
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Description (incidence and severity):
No change due to the test substance was observed.
A non-delivering animal was found in one patient in the 2 mg / kg group. which had one implantation.(it wa know at necropsy, male of the mating). It was thought that it was attributed to males, as the size of both sides of the epididymis was seen. However, it seems to be spontaneous as a change seen in the stone, from the number of examples of its occurrence-emergent change. In addition, one male in the 10 mg / kg group also had testes and fine small one side of the upper body was seen, but the mating partner normally got pregnant, delivered. The gestation period, the number of corpus luteums, the number of landings, the implantation rate, the birth rate,there was no significant difference between the administration group and the control group. Also, for any mother animal no abnormalities were found in the parturition condition and nursing behavior.
Key result
Dose descriptor:
NOAEL
Effect level:
ca. 2 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: no changes attributed to the test substance
Key result
Dose descriptor:
LOEL
Effect level:
ca. 2 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: no changes attributed to the test substance
Key result
Critical effects observed:
no
System:
male reproductive system
Key result
Critical effects observed:
no
System:
female reproductive system
Clinical signs:
no effects observed
Description (incidence and severity):
In the 10 mg / kg group, sex ratio (male/(male + female), male/female) in the total number of babies and births, there was a significant difference in total group number.
No abnormalities attributable to any of the pups were observed.
An increase of number of stillborns, decrease of live birth index and viability index on day 4 were observed.
Dermal irritation (if dermal study):
not examined
Mortality / viability:
mortality observed, treatment-related
Description (incidence and severity):
In the 10 mg / kg group, low birth rates, 4 day surviving infants and 4 day survival rates were observed.
In the 2 mg / kg, a low 4-day survival rate was also seen in one animal. However, it was a change in only one case, and the 4-day survival rate in other mother animals of the same group was about the same value as that of the control group. - accidental change.
Body weight and weight changes:
not specified
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Histopathological findings:
not specified
Other effects:
effects observed, non-treatment-related
Description (incidence and severity):
Jaundice was noted in 1 dead offspring each in the 2 and 10 mg/kg groups.
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
ca. 2 mg/kg bw/day (nominal)
Sex:
not specified
Basis for effect level:
viability
other: An increase of number of stillborns, decrease of live birth index on day 4.
Key result
Dose descriptor:
NOEL
Generation:
F1
Effect level:
ca. 2 mg/kg bw/day (nominal)
Sex:
not specified
Basis for effect level:
viability
other: An increase of number of stillborns, decrease of live birth index on day 4.
Key result
Critical effects observed:
not specified
Key result
Reproductive effects observed:
no
Conclusions:
The test substance had no effects on the reproductive parameters nor maternal behavior of parents animals and does not cause any abnormalities on the offspring. Two mg/kg/bw was evaluated as the NOAEL and NOEL value.
Executive summary:

Thiosemicarbazide was tested by oral route to rats in the doses 0, 0.4, 2 and 10 mg/kg/bw. Dosing period was 42 days far males, for satellite females 42 days and for females 14 days before mating to day 4 of lactation. There were used 12 male animals per group (5 for recovery), 12 females animals and 5 satellite females.According to the data obtained from the Nagtional Institute of Technology and Evualtion, the tested Thiosemicarabzide does not have effects to the reproductive parameters such as estrous cycle, copulation index, fertility index, delivery index, gestation length, numbers of corpora lutea or implantations, implantation index, gestation index, parturition, or maternal behavior of parents animals in tested concentrations. No general toxicity to maternal animals was observed. However, a significant reduction in the number of live offspring on day 4, as well live birth index and viability index on day 4 was noted. At the necropsy, there was noted jaundice in 1dead offspring, each in 2 and 10 mg/kg/bw.However the relation to the test substance was not clear, since no similar changes were noted in any other offspring. No changes attributed to the test item were noted in the number of offspring or live offspring at birth, sex ratio, clinical signs, external features, or body weights. Based on this data, to NOAEL and NOEL value was assigned the value 2.0 mg/kg/bw.

Endpoint:
one-generation reproductive toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP and Guideline followed
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Test material information:
Composition 1
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material:Cold and dark place
Species:
rat
Strain:
Crj: CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: H Tree-Jarls. Riva One Day Koki Center
- Age at study initiation: 9 weeks old
- Acclimation period: 6 days including quarantine
Route of administration:
oral: gavage
Vehicle:
water
Remarks:
Purified
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
MALES: 42 days
FEMALES: 41 - 47 days (from 14 days before mating to day 4 of lactation)
FEMALES (satellite) : 42 days
Frequency of treatment:
Daily
Dose / conc.:
0 mg/kg bw/day (nominal)
Dose / conc.:
0.2 mg/kg bw/day (nominal)
Dose / conc.:
1 mg/kg bw/day (nominal)
Dose / conc.:
5 mg/kg bw/day (nominal)
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
RECOVERY
Dose / conc.:
5 mg/kg bw/day (nominal)
Remarks:
RECOVERY
No. of animals per sex per dose:
MALES: 12 (5 for recovery) in each group
FEMALES: 12 n each group
SATELLITE FEMALES: 5 n each group
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Based on the results of range finding study.

Doses of 0, 0.016, 0.08, 0.4, 2 and 10 mg / kg. It was set with reference to the result of the dose setting test conducted at the test facility. "14-day repeated dose toxicity by oral administration of Hydrazinecarbothioamide in rats Test "
Dosing perid: 14 days

Death occured in one animal of three in total in the doses of 10 mg/kg and thsid ead animal had cyanosis and pulmonary edema The decrease of locomotor activity, respiration rate, body waight in males. Males appeared to had incomplete eyelid opening. The value of ALT decreased and T-Cho increased in t males animals. Females had blakish region of mucosa in the glandular stomach. Increase of weight of kidney R and decrease of weight of Thymus Al,, R was seen in both sexes. These abnormalities are conected with doses of 10 mg/kg .In the doses of 2 mg/kg Decrease of thymus A,R and increase of Kidney R was evaluated . Because of these findings, the selected dose for the study were doses: 0,0.2, 1 and 5 mg/kg/day.
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes, (loss of hair, scab formation exudate on males and reddish tear, staining around nose and moth, staining lower abdomen, unclearance of placenta, nursing loss, udevelopment of nipple, litter loss, loss of hair in females, tremors,twich or convulsion, stereotypic behaviour and abnormal behaviour). In amles during the 6 weeks and 2 revcovery week and in females during 6 weeks , two recovery week and post partum day 4 observation were made.

DETAILED CLINICAL OBSERVATIONS: Yes (abnormalities, vocalization, ease of removal, musele tone, subnormal temperature, piloerection, staining hair, unkempt hair, paleness, reddening, cyanosis, lacrimation, exophthalmos, pupikkary size, salivation, secretion)

BODY WEIGHT: Yes, In males day 1,3,7,14,21,28,35 and 42 - administration period, and during recovery period on day 1,3,7 and 14.
In females, before copluation on day 1,3,7 and 14, during gestion on day 0,7,14,17 and 20 and postapartum period on day 0 and 4.
Satellite females, during administration period on day 21,28,35,42 and recovery days 1,3,7 and 14.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):Yes, In males during administration perido on day 3,7,14,21, 28, 35 and 42. Recovery period on day 3,7 and 14.
In females before copulation on day 3,7 and 14, during gestion on day 7,14,17 and 20 and on post partum day 4. Satellite females during administration period on day 21,28,35,42 and recovery days 1,3,7 and 14.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Not specified

OTHER:

HAEMATOLOGY: Yes (RBC, MBC, Hb, Ht, MCV,MCH,MCHC,Platalet, Reticulo, PT and APTT, Differentiation of leucocyte,)

CLINICAL CHEMISTRY: Yes (AST, ALT, ALP, ChE, gama-GTP,T-Cho, TG, Glucose, T-Protein, Albumin, A/G ratio, BUN, Creatine, T-Bil, Ca, IP, Na, K,Cl )

NEUROBEHAVIOURAL EXAMINATION: Yes ( posture, motor activity, respiration, lid closure, gait, touch response4, pinna and pain response, pupillary effect and air righting reflex) Observations: Males in the week 6 and female post partum day 4.
Estrous cyclicity (parental animals):
Yes (Cycle length, number of females with normal/abnormal activity, abnormal cyclicity ratio)
Sperm parameters (parental animals):
No data
Litter observations:
PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
pups born, stillborns, live pups at birth, live pups at the end of observations on postnatal day 0 and number of live pups on postnatal day 4, live birth index, viability index


Postmortem examinations (parental animals):
SACRIFICE
- Male animals: Males, day 43 of treatment and 15 of recovery
- Maternal animals: Females, day 5 of lactation

GROSS PATHOLOGY: Yes (forestomach, glandural stomach, jejunum, liver-whitish region, kidney-pelvic dilatation, Epididymis-whitis region, Thymus-small, Adrenal-enlargement, Skin-loss of hair)
ABSOLUTE ORGAN WEIGHTS:Yes (liver, heart, kidney, testis, epididymis, brain, spleen, Thymus, Adrenal, body weight)
RELATIVE ORGAN WEIGHTS: Yes (liver, heart, kidney, testis, epididymis, brain, spleen, Thymus, Adrenal, body weight)

HISTOPATHOLOGY: Yes (Trache, lung-abnormalities and aggregation of foamy cells, Forestomach-abnormalities and squamous epithelial cyst, Grandular stomach-abnormalities, Duodenum-abnormalities, Jejunum-abnormalities, diverticulum, Mineralization in Peyers patches, Ileum, Cecum, Colon and Rectum for abnormalities, Liver-abnormalities, focal necrosis of hepatocytes, nicrogranuloma, Herat-abnormalities and focal myocarditis, Kidney-abnornmalities, pelvic dilatation unilateral, Solitary cyst in medula and vacuolization of tubular epithelium and urinary bladder for abnormalities, testis-abnormalities and focal atrophy of seminiferous tubules, bilateral, Epididymis-abnormalities and lumphocyte infiltration, SProstate-abnornmalities and lumphocyte infiltration, Seminal vesicle, Ovary-abnormlities and increased follicular atresia, Uterus-abnormalities and endometrial atrophu, vagina-abnormalities and mucification of epithelum, Celebrum, Celebrium, Pons, Spinal cord, Sciatic nerve, Bone narrow, Axillar lymp node, Mesenteric lymoph node, spleen, thymus-abnormalities and atrophy, pituitary gland, thyroid, parathyroid, adrenal and skin for abnormalities.).
Postmortem examinations (offspring):
SACRIFICE
- Day 4 after birth
Reproductive indices:
Yes (number of mated pairs, copulated pairs, pregnan females, copulation index, fertilization index, conception index and pairing days until copulation)
Offspring viability indices:
Viability indices on day 4 were observed. Survival, litter loss, sex ratio and number of stillborns were observed.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
No abnormalities were detected in detailed clinical observations. In the concentration of 5 mg/kg/bw was observed in the males loss of fur, crust formation, exudate,in the females groups Soiled perineal region, Reddish tear (5,3 dams with total litter loss), Loss of fur.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
not specified
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
A temporary decrease in food consumptions was observed in males given 5 mg/kg.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not specified
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
Males: Increase of T-Cho and an increase in total cholesterol in the doses of 5 mg/kg/bw
Urinalysis findings:
not examined
Behaviour (functional findings):
not specified
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Decreases in absolute and relative weights of the thymus in males given 5 mg/kg were observed at the end of the administration period.The recovery of the effects on the thymus was noted at the end of the recovery period.
Gross pathological findings:
not specified
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Males: Atrophy of the thymus (small size) was observed in 5 mg/kg/bw at the end of the administration period.
Females (5, 3 dams with total litter loss): Small size and atrophy of the thymus, Vacuolization of tubular epithelium in the kidney, Enlargement of the adrenal, Atrophy of the spleen.
Histopathological findings: neoplastic:
not specified
Other effects:
not examined
Reproductive function: estrous cycle:
no effects observed
Reproductive function: sperm measures:
no effects observed
Reproductive performance:
effects observed, non-treatment-related
Description (incidence and severity):
An increase in stillbirth and a decrease in viability indices on days 0 and 4 after birth were observed in the next generation of the group of 5 mg/kg caused by nursing failure of the dams.
Key result
Dose descriptor:
NOAEL
Effect level:
ca. 1 mg/kg bw/day (nominal)
Sex:
female
Basis for effect level:
other: Since nursing failure of the dams, an increase in stillbirth and a decrease in viability indices on days 0 and 4 after birth were observed in the next generation of the group of 5 mg/kg.
Key result
Dose descriptor:
LOEL
Effect level:
ca. 1 mg/kg bw/day (nominal)
Sex:
female
Basis for effect level:
other: Since nursing failure of the dams, an increase in stillbirth and a decrease in viability indices on days 0 and 4 after birth were observed in the next generation of the group of 5 mg/kg.
Key result
Critical effects observed:
not specified
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
A increase of total litter loss, an increase of number of stillborns, decrease of live birth index and viability index on day 4.
Dermal irritation (if dermal study):
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not examined
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Histopathological findings:
not specified
Other effects:
not examined
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
ca. 1 mg/kg bw/day (nominal)
Sex:
not specified
Basis for effect level:
other: Since nursing failure, observation on next generation F2 (A increase of total litter loss, an increase of number of stillborns, decrease of live birth index and viability index on day 4.)
Key result
Dose descriptor:
NOEL
Generation:
F1
Effect level:
ca. 1 mg/kg bw/day (nominal)
Sex:
not specified
Basis for effect level:
other: Since nursing failure, observation made in next geneartion F2(A increase of total litter loss, an increase of number of stillborns, decrease of live birth index and viability index on day 4.)
Key result
Critical effects observed:
not specified
Key result
Reproductive effects observed:
no
Conclusions:
No abnormalities were detected in parental animals nor in the offsprings in tested concentrations of Thiosemicarbazide.One mg/kg was evaluated as the NOAEL and NOEL value because of the decrease in viability index and an increase in stillbirth on day 0 and 4 in the next generation in the group of 5 mg/kg
Executive summary:

Thiosemicarbazide was tested by oral route to rats in the doses 0, 0.2, 1 and 5 mg/kg/bw. Dosing period was 42 days far males, for satellite females 42 days and for females 14 days before mating to day 4 of lactation. There were used 12 male animals per group (5 for recovery), 12 females animals and 5 satellite females.According to the data obtained from the Nagtional Institute of Technology and Evualtion, an increase in whole litter loss and the decrease in viability index on postnatal day 4 in the next generation caused by nursing failure of the dams were observed in the groups of 5 mg/kg. No general toxicity to the maternal aniamls was observed.The tested Thiosemicarabzide does not have effects to the reproductive parameters such as estrous cycle, copulation index, the number of corpora lutea, number of implantation sites or gestation length in parental animals in tested concentrations. Based on this data, to NOAEL and NOEL value was assigned the value 1 mg/kg/bw.

Justification for classification or non-classification

The substance can not be classified for toxicity to reproduction according to CLP Regulation (EC) no. 1272/2008 due to insufficient data.