Registration Dossier

Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
one-generation reproductive toxicity
Remarks:
based on test type (migrated information)
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
27.02.2008 - 25.09.2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study performed according to the guideline.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008
Report Date:
2008

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EU Method B.34 (One-Generation Reproduction Toxicity Test)
Qualifier:
according to
Guideline:
OECD Guideline 415 [One-Generation Reproduction Toxicity Study (before 9 October 2017)]
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Details on test material:
The appearance of Potassium permanganate was dark violet - purple crystalline powder with bronze lustre.

Potassium permanganate has been used as a surrogate for sodium permanganate where data are not available. Read-across from potassium permanganate to sodium permanganate is appropriate from the toxicological point of view as the most toxicologically relevant part of the substances is the same (permanganate). The contribution of the sodium/potassium ions to the toxicity of the respective substances is likely to be minimal. The toxicity of both substances is therefore likely to be very similar and will be dominated by local (site of contact) irritant/corrosive effects and systemic toxicity due to the absorption of manganese ions. This toxicophore similarity is adequate justification for waiving the conduct of specific studies with sodium permanganate and the dossier reflects this waiving proposal by including summaries of read-across studies where appropriate.

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
The rats were supplied from SPF breeding, VELAZ s.r.o., Kolec u Kladna, Czech Republic, RCH CZ 21760118. Age at the start of the study was 5-6 weeks for males, 9 weeks for females and all animals were acclimitised for a minimal of 5 days. 40 males and 100 females (10 males and 25 females per group) were used in the test. Animals were fed a complete pelleted diet for rats and mice and were allowed unrestricted access to drinking water. Animals were housed in an SPF animal room, 2 rats of the same sex in one plastic cage (40x25x20 cm) containing sterlised clean shavings of soft wood. Animals were housed in a controlled environment, in which optimal conditions were considered to be approximately 15 air changes per hour, a room temperature of 22±3oC, a relative humidity of 30-70% and a 12-hour light/ 12-hour dark cycle.

Administration / exposure

Route of administration:
oral: drinking water
Type of inhalation exposure (if applicable):
other: oral application using a stomach tube
Vehicle:
water
Details on exposure:
The concentrations of solutions in all three dose levels were adjusted to ensure the administration of 1mL per 100 g of body weight. The application form (test substance in water for injection) was prepared daily just before administration. The vehicle control group was administered water for injection in the same volume. The application form of the test substance was prepared daily before administration; these solutions were mixed for 10 minutes by magnetic stirrer. The treated and control groups were administered daily by gavage. Oral administration was chosen on the request of the sponsor. The animals were treated 7 days per week at the specified time (08.00 - 10.00).
Details on mating procedure:
During the mating period, one male with two females were housed together in one cage. Females that mated were individually housed in labelled cagesand the offspring was kept with mother until weaning. Vaginal smears were prepared daily during the mating period (until the presence of spermatozoa was noted). Smears were carried out in the female's morning every day during mating procedures. The smears were stained and the presence of sperm was watched, day 0 of pregnancy was defined as the day when the sperm are found.
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
No details given.
Duration of treatment / exposure:
The treatment period for the males was 13 weeks (10 weeks of pre-mating and 3 weeks of mating period). The treatment period of females - mothers was at least 8 weeks (2 weeks of pre-mating, at least 1 day of mating, 3 weeks of pregnancy and 3 weeks of lactation).
Frequency of treatment:
The test substance was administered at the following dose levels:
-the lowest dose: 20 mg/kg body weight/day
-intermediate dose: 80 mg/kg bodyweight/day
-the highest dose: 320 mg/kg bodyweight/day
The animals were treated 7 days per week at the specified time (8.00 - 10.00am).
Details on study schedule:
The test substance was administered as water solution using a stomach tube; oral application of rats was made daily. The volume administered was adjusted to 1 mL per 100g of bodyweight in all doses. The four groups of animals were included in the study - 3 treated groups (doses 20, 80, 320 mg/kg/bw/day) and one control group (vehicle only).
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
20 mg/kg/bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
80 mg/kg/bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
320 mg/kg/bw/day
Basis:
actual ingested
No. of animals per sex per dose:
Each group consisted of 10 males and 25 females.
Control animals:
yes, concurrent vehicle
Details on study design:
No further information provided.
Positive control:
Not included in this study.

Examinations

Parental animals: Observations and examinations:
Bodyweight:
Males - the first day of administration and then weekly.
Females - the first day of administration and then weekly, during pregnancy: 0, 7th, 14th and the 21st day. Also during lactation: 1st, 4th, 7th, 14th and 21st day.
Pups - litters: 1st, 4th, 7th, 14th and 21st day of life.
Pups - individually: 21st day of life.

Food consumption:
-during premating period weekly.
-during pregnancy and lactation - on the same days as body weight (daily consumption was counted).

Mortality control: daily.
Health condition control: daily-during the acclimitisation and the experimental part.
Clinical observations: daily-during the administration period.

Laboratory examinations:
-vaginal smears: daily in the mating period.
-pathological examination: males - after ending of the mating period, mated females without delivery (nonpregnant or aborted) - 25th day after mating, parental females and pups - on the 21st day of lactation.
-biometry of organs: during necropsy.
Oestrous cyclicity (parental animals):
Estrous cyclivity was not determined.
Sperm parameters (parental animals):
Sperm parameters were not determined.
Litter observations:
Reproduction parameters relevant to pups (number of pups, bodyweight, sex or vitality) was also recorded.
Postmortem examinations (parental animals):
Parental males were euthanised at the end of the mating period - 93rd day of study (after 92 days of application). Parental females were euthanised on the 21st day of lactation. Mated females without delivery (nonpregnant or aborted) were euthanised on the 25th day after mating. Animals were then macroscopically examined for any structural abnormalities or pathological changes with special attention to the organs of the reproductive systems. All macroscopic abnormalities were recorded.
Postmortem examinations (offspring):
Dead pups or pups euthanised before the 14th day of lactation were sexed and externally examined; the stomach was examined for the presence of milk. Pups found dead or euthanised on or after the 14th day of lactation were sexed and subjected to external examination of the cranium, and to macroscopic examination of the thoracic and abdominal tissues and organs. All macroscopic changes were recorded.
Statistics:
The ANOVA test at a significance level of 0.05 was used for the statistical analysis. This analysis was used for the results of body weight and biometry of organs. The control group with vehicle was compared to the three treated groups.
Reproductive indices:
The absolute weight of testes, epididymides, prostate gland and pituitary gland were recorded in males and the absolute weight of ovaries, uterus (incl. uterine tube and cervix) and pituitary gland were recorded in females. Afterwards the somatic indexes - SI (= relative weight of organ) were computed according to the following formula: SI - weight of organ x 100/bodyweight.
Offspring viability indices:
Each litter was examined as soon as possible after delivery and the number and sex of pups, still births, live births and presence of gross anomalies were recorded. Changes of physical condition and behaviour anomalies were recorded. Development of pups (ability of sucking, growth of hair, teeth and dugs, opening of eyes and ears) was observed in the reported days.

Results and discussion

Results: P0 (first parental animals)

General toxicity (P0)

Clinical signs:
effects observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
effects observed, treatment-related
Reproductive function: sperm measures:
not examined
Reproductive performance:
effects observed, treatment-related

Details on results (P0)

The application of the test substance at the dose level of 320 mg/kg/day had a negative influence on clinical status of both sexes (more marked in males) and caused death of one nonpregnant female. The negative effect of the treatment on growth of males and on food consumption of both sexes was also recorded. Toxicity of the test substance at the highest dose level was also confirmed during necropsy and histological examination of males and females (numerous bleeding erosions of stomach or duodenum). Histopathological examination of the reproductive system of parental males showed high incidence of affected males at the 320 mg/kg/day dose level. Detailed examination of testes revealed various damage of spermatogenesis. Significant decreased weight of the prostate gland was recorded also at the highest dose level and could reflect changes in testicular function. Examination of reproductive system of parental females showed a relatively high incidence of findings at all dose levels including the control group but without dose dependence. There was no significant pathological affections to indicate damage of reproductive female organs at any of the dose levels.
The number of pregnant females, the accompanying fertility index and conception index were all lowered at the highest dose level. This result showed decreased ability of the animals to achieve a pregnancy. Duration of pregnancy in treated groups was similar to the control group. The gestation index was relatively well-balanced at all dose levels. Administration of the test substance did not cause significant postnatal loss and pre-weaning loss (viability index and weaning index). The other observed parameters (percentage of live males and females - at first check of litter and also at weaning) were unaffected and were in the normal range.

Effect levels (P0)

open allclose all
Dose descriptor:
NOAEL
Remarks:
reproduction
Effect level:
80 mg/kg bw/day
Sex:
male/female
Basis for effect level:
other: Marked reduction in litter number at 320 mg/kg bw/d
Dose descriptor:
NOAEL
Remarks:
parental
Effect level:
80 mg/kg bw/day
Sex:
male/female
Basis for effect level:
other: Toxicity, bodyweight effects, gastric irritation, pathology of the reproductive tract

Results: F1 generation

General toxicity (F1)

Clinical signs:
effects observed, treatment-related
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Sexual maturation:
effects observed, treatment-related
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
effects observed, treatment-related
Histopathological findings:
effects observed, treatment-related

Details on results (F1)

The number of pups, sex ratio, average weight of litter and average body weight of pups was unaffected and well-balanced with the control group. Presence of macroscopic abnormalities - oedematous appearance of brain was recorded in pups at the highest dose level. Statistical examination showed significant increased absolute and relative weight of the brain at the highest dose level. During histological examination, the high incidence of vacuolisation of cell nuclei in the cortex and hippocampus was recorded at all dose levels. This change was more intensive at the middle and high dose levels. Observation of development (until 14 day after birth) showed late opening of eyes at the middle and highest dose level. The change was marked at the highest dose level and could be related with biometrical and histopathological findings. Sporadic but significant pathologic changes of stomach and genital organs was recorded in pups at the highest dose level - missing or reduced testis and epididymis, congested stomach mucosa and chyme with blood. These changes could be considered as changes of toxicological significance.

Effect levels (F1)

Dose descriptor:
LOAEL
Remarks:
developmental
Generation:
F1
Effect level:
20 mg/kg bw/day
Sex:
male/female
Basis for effect level:
other: Brain pathology in all treated groups

Overall reproductive toxicity

Reproductive effects observed:
not specified

Any other information on results incl. tables

OBSERVATION OF PARENTAL MALES       

MORTALITY

There were no unscheduled deaths during the study.

BODY WEIGHT INCREMENT       

The body weights in the dose levels 20 and 80 mg/kg/day was relatively well balanced with the control group until the 6th week of application and then it slightly decreased. Markedly lower body weight was recorded at the 320 mg/kg/day dose level since the 1st week and at the end of the study (11th week) the body weight increment was negative (animals lost weight). Body weights of males at the highest dose level were statistically significantly decreased at the end of the study.

FOOD CONSUMPTION

Food consumption in the 20 and 80 mg/kg/day was well-balanced with the control group until the 4th week of application, and from the 5th week to the end of the study, the consumption was slightly decreased. At the highest dose level (320 mg/kg/day) the food consumption was decreased from the 3rd week of the application period to the end of the study. The change was marked since the 8th week.

HEALTH CONDITION

The health condition of the control males and males treated by the test substance at the dose level 20 mg/kg/day was very good during the whole study. No signs of diseases or toxic effect owing to application of the test substance was found, one male from the control group showed diarrhoea and one male from the lowest dose group showed red secretion from the nostrils. At the 80 mg/kg/day dose level, s

poradic dyspnoea, red secretions and salivation were recorded more often in the control and lowest dose level groups. These changes were incidental and did not affect all animals in the group. The health condition of animals in the 320 mg/kg/day dose group was not good from the 1st week of the application period. The changes recorded included: dyspnoea, decreased activity, red secretion around the nose or eyes, rigidity, piloerection and salivation, which were recorded in most of the animals.

MACROSCOPIC EXAMINATION

Examination of the external surface of the body or the thoracic cavity revealed no changes. Sporadic changes of abdominal cavity were recorded in the control group and at the 20 and 80 mg/kg/day dose levels. More frequent changes were recorded at the 320 mg/kg/day dose level. Reduced testes, prostate glands, epididymides or seminal vesicle were observed sporadically. All males at this dose level showed marked changes in the stomach, blood erosions of mucosa and the same males showed erosions of the duodenum, dilation of the stomach or changes in mucous membrane or liver affections. These treatment-related findings were considered as changes with toxicological significance.

BIOMETRY OF REPRODUCTIVE ORGANS

Absolute and relative weights of all observed organs were relatively well-balanced at the 20 and 80 mg/kg/day dose levels compared to the control group. Animals at the 320 mg/kg/day dose level showed a decrease of absolute weight of the prostate gland with statistical significance. Slight decreases in absolute weight of testes and epididymides (without statistical significance) was recorded also at the highest dose level.

HISTOPATHOLOGICAL EXAMINATION

The incidence of histopathological affects of the reproductive system in the male genital tract was sporadic in the control group and the lower dose level. Marked chamges were recorded at the highest dose level. Detailed examination of testes revealed various damage of spermiogenesis in 1-3-0-7 males, atrophy of germinative epithelium in 3-0-3-1 males and atrophy or decreased quantity of Leydig cells in 0-1-0-1 males. Damage of spermiocytes (decreased number or presence of necrotic cells) was observed in epididymides of 0-1-0-3 males, vascular dystrophy in 0-2-0-0 and inflammation in 1 -0 -2 -0 males. Signs of inflammation in the prostate gland (oedema of interstitium, lymphocyte infilration or inflammation) were recorded in 2-2-5-3 males. In the digestive system, the stomach, forestomach and duodenum were affected. Erosions, ulcerations and haemorrhage of stomach mucosa or submucosa were described in 0-0-1-8 males. Inflammation (including inflammatory infiltration of mucosa and/or submucosa) was diagnosed in the stomach of 0-0-1-10 males, in the forestomach of 0-0-1-6 males and in the duodenum of 0-0-0-2 males.

Administration of the test substance Potassium permanganate affected clinical status, growth of parental animals, biometrics and microscopic structure of reproductive male organs and development of pups. In males that received the highest dose level, these effects manifested in significant damage of spermatogenesis with a negative effect on fertility. These changes resulted in a decreased ability of animals to achieve a pregnancy. Application of the test substance at the highest and middle dose levels had a negative developmental effect on pups (pathological changes of brain and late opening of eyes).

OBSERVATION OF PARENTAL FEMALES

MORTALITY

There were no unscheduled deaths at the 20 and 80 mg/kg/day dose levels and in the control group during the study. One nonpregnant female at the highest dose level died in the first week after mating.

BODY WEIGHT: PRE-MATING PERIOD

Average body weight increments of females at the 320 mg/kg/day dose level were markedly lower than in the control group at the end of the 1st week of application (tabescence of animals)

PREGNANCY       

Females without parturition (non pregnant or aborted females) were not included in evaluation of body weight during pregnancy. Body weight increment of pregnant females at the 20 mg/kg/day dose level was relatively well-balanced compared to the control group. A slight decrease of increment was recorded at the 320 mg/kg/day dose level (only at the end of the 1st and 2nd week of pregnancy) and at the 80 mg/kg/day dose level (at the end of the 2nd week of pregnancy).

LACTATION

Only mothers (females with live pups) were included in the evaluation of body weight increment during the lactation period. Until the 14th day of lactation all groups of animals (treated and control) showed an increase of body weight (positive increment). The body weight increment of treated groups was relatively well-balanced with the control group. Lactation (1st to the 4th day) was slightly decreased at the 20 and 320 mg/kg/day dose levels. A marked decrease in body weight was recorded at the end of lactation (from the 14th to the 21st day of the study) in all groups of animals (treated and control). The body weight increment of all groups was negative. Marked tabescence was recorded at the 80 mg/kg/day dose level, whereas the tabescence of animals following the 320 mg/kg/day dose level was not so marked.

FOOD CONSUMPTION: PRE-MATING PERIOD

Average food consumption at the 20 and 80 m/kg/day dose levels was well-balanced with the control group during the whole pre-mating period. Food consumption of females at the 320 mg/kg/day dose level was lower than in the control group at the end of the 1st week of application.

PREGNANCY

Females without parturition (non-pregnant or aborted females) were not included in the evaluation of food consumption during pregnancy. Average food consumption in all treated groups during pregnancy was relatively well-balanced with the control group.

LACTATION

Only mothers (females with live pups) were included in the evaluation of food consumption during the lactation period. Average food consumption of all treated groups was lower than in the control group during the whole lactation period. A more marked decrease was registered in the 20 and 320 mg/kg/day dose levels.

HEALTH CONDITION

The health condition of control females and females at the 20 and 80 mg/kg/day dose levels was very good during the whole study. No signs of diseases or toxic effects related to application of the test substance were recorded. One female from the control group showed piloerection (only in the 6th week). At the lowest dose level, one female showed sporadic dyspnoea (in the 5th week) and two females showed red secretions around the nose (1st and 3rd week). Alopecia was also recorded in one female from the 1st-7th week of application. At the middle dose level, only one female showed red secretions around the nose from the 2nd-4th week. At the highest dose level, dyspnoea was recorded in females (in week 1 - 3 females, in week 2 - 2 females, in week 4 - 1 female and in week 7 - 2 females). In the absence of a dose-related response this was considered to be incidental.

MACROSCOPIC EXAMINATION

Examination of the external surface of the body revealed no changes. In the thoracic cavity no changes were observed. No changes in the abdominal cavity were recorded at the 320 mg/kg/day dose level. Nine females at this dose level showed bleeding erosions of stomach mucosa and one female also showed blood in the duodenum or liver effects. These findings with treatment-related distribution were considered as changes with toxicological significance. Dilation of the uterus was more numerous at the highest dose level, and can be related to the increased number of nonpregnant females at this dose level.

BIOMETRY OF REPRODUCTIVE ORGANS

Nonpregnant females and females with abortion were not used for calculation of means and evaluations of organ weight. Relative and absolute weight of all measured organs was relatively well-balanced (without statistical differences) at all the dose levels.

HISTOPATHOLOGICAL EXAMINATION

In the digestive system the stomach and forestomach was affected. Erosions, ulcerations and haemorrhage of stomach mucosa or submucosa was described in 0-0-0-5 females. Inflammation (inclusive inflammatory infiltration of mucosa and/or submucosa) was diagnosed in the stomach of 0-0-0-9 and in the forestomach of 0-0-0-3 females. In the reproductive system, the ovaries and uterus were markedly affected. Follicular cysts of ovaries were recorded in 20-16-22-13 females and cellular hyperplasia of strome in 2-0 -2-3 females. In the uterus, cellular hyperplasia of the endometrium was found in 13-15-4-9 females, and hydrometra in 4-0-1-7 females. Degenerative changes (atrophy of endometrium, extinction of endometrial glands, fibrosis of endometrium or atrophic epithelium in vagina) were recorded in 3-0-3-7 females. Cysts in the pituitary gland were recorded in 0-1-1-0 females.

OBSERVATION OF PUPS

NUMBER AND SEX RATIO OF PUPS

The total number of pups was decreased wth the dose level and the lowest number was found at the 320 mg/kg/day dose level. The number of pups per litter was relatively well balanced in all treated groups. Slightly lowered numbers but without statistical significance was recorded at the 320 mg/kg/day dose level.

BODY WEIGHT

The average body weight of pups in all treated groups was well balanced with the control group and had an increasing trend during the whole lactation period.

DEVELOPMENT OF PUPS

Death of pups was minimal and similar in all groups:

- up to the 4th day after birth, 3 pups died in the control group (from 1 mother), 3 pups from the 80 mg/kg/day dose group (from 3 mothers) and 4 pups from the 320 mg/kg/day (from 2 mothers).

- up to the 7th day after birth, no other pups died.

- up to the 14th day after birth, only one pup died at the 320 mg/kg/day dose level

- up to the 21st day after birth, no other pups died.

There were no differences in the development of pups observed at the 320 mg/kg/day dose level. Observation of the opening of eyes (until 14 days after birth) showed late opening at the 80 mg/kg/day dose level (2 litters from 16) and at the 320 mg/kg/day (3 litters from 10).

MACROSCOPIC EXAMINATION

The pathological examination was performed in all pups. All newborn pups that had died during the study were examined for the presence of air in the lungs and divided into still birth and live birth pups. Presence of macroscopic abnormalities could not be detected in dead pups with autolysis. No pathologic findings was recorded in the control group and at the 20 and 80 mg/kg/day dose levels. Sporadic pathological findings were recorded at the 320 mg/kg/day dose level: missing testes and epididymides (one pup), one testis reduced (one pup) and stomach mucous membrane congested and chyme with blood (two pups).

EXAMINATION OF BRAIN

The absolute weight of the brain was increased in all treated groups with statistical significance at the 320 mg/kg/day dose level. The relative weight of the brain was increased in all treated groups depending on dose level and statistical significance at the 320 mg/kg/day dose level. During histopathological examination the vacuolisation of cell nuclei in the cortex and/or hippocampus was recorded at all dose levels but it was more marked in treated groups. Only mild vacuolisation was recorded in 3 control pups. The number of pups with vacuolisation in the brain was well-balanced in all treated groups but marked vacuolisation was recorded at the middle and highest dose level.

REPRODUCTION PARAMETERS

All females (in all groups) were mated so that the number of females mated was identical in all groups. The number of pregnant females and accompanying number of mothers bearing live pups was markedly lower at the highest dose level. The duration of pregnany of treated groups was similar to the control group. A significant decrease in the fertility index was detected only at the highest dose level. The percentage of postnatal loss was slightly increased at the middle dose level. All other calculated parameters were relatively well-balanced.

Applicant's summary and conclusion

Conclusions:
Administration of the test substance Potassium permanganate affected clinical status, growth of parental animals, biometrics and microscopic structure of reproductive male organs and development of pups. In males that received the highest dose level, these effects manifested in significant damage of spermatogenesis with a negative effect on fertility. These changes resulted in decreased ability of animals to achieve a pregnancy. Application of the test substance at the highest and middle dose levels had a negative developmental effect on pups (pathological changes of brain and late opening of eyes).

The NOAEL for reproduction was established as 80 mg/kg bodyweight/day. The NOAEL for development was established as 20 m/kg bodyweight/day.
Executive summary:

Administration of potassium permanganate in a one-generation reproductive toxicity study was found to have effects at the highest dose level of 320 mg/kg bw/d. Signs of toxicity, poor general condition, reduced bodyweights and food consumption were seen in this group. Pathology revealed marked irritant effects on the gastric mucosa in both sexes and effects on the male reproductive organs. The number of litters was markedly reduced at 320 mg/kg bw/d, however litter size was unaffected by treatment. Bodyweights and survival of pups was similar in all groups, however eye opening was delayed in pups at the top dose level. Necropsy of pups revealed increased brain weight at 320 mg/kg bw/d and histopathology showed increased vacuolisation of cortical/hippocampal cells in all treated groups.