polymeric zinc propylenebis(dithiocarbamate);propineb (ISO)

Toxicity to reproduction

Administrative data

Endpoint:

two-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2008/6/26 to 2010/6/04

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)

Version / remarks:

2001

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.3800 (Reproduction and Fertility Effects)

Principles of method if other than guideline:

The exception is that the homogeneity and stability of the test substance in the diet were verified after the study was completed, due to unanticipated challenges associated with developing the analytical method. This is not believed to have had an effect on the outcome or interpretation of the study, since the results verified the homogeneity and stability of the test substance in the feed, under the conditions that were used in this study.

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

polymeric zinc 1,2-propylenebis(dithiocarbamate)

EC Number:

620-365-5

Cas Number:

9016-72-2

IUPAC Name:

polymeric zinc 1,2-propylenebis(dithiocarbamate)

Test material form:

solid

Remarks:

Powder

Test animals

Species:

rat

Strain:

Wistar

Remarks:

Han CRL

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Inc. Raleigh, North Carolina
- Age at study initiation: (P) 8-9 weeks old
- Weight at study initiation: (P) Males: mean value range: 214.2 – 258.7 g; Females: mean value range: 127.4 – 175.4 g;
- Housing: Individual hanging stainless steel cages with deionized cage board in the bedding tray. During the gestation and lactation phases, individual dams and their litters, and F1 and F2 pups were housed in polycarbonate cages with ground corn cob bedding (Bed-OCOBs).
- Diet: Purina Mills Certified Rodent Diet 5002 meal, St. Louis, MO (USA)
- Water: Water from the municipal supply, ad libitum
- Acclimation period: one week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-26°C
- Humidity (%): 30-70%
- Air changes (per hr): at least 10.67 air changes per hour
- Photoperiod (hrs dark / hrs light): Alternating 12-hour light and dark cycles
IN-LIFE DATES: The study was initiated on July 7th 2008 and the in-life phase was completed on April 30th 2009.

Administration / exposure

Route of administration:

oral: feed

Vehicle:

acetone

Details on exposure:

Diet preparation and analysis:
The test substance was dissolved in acetone and then mixed with the feed. Treated diet was mixed at room temperature; aliquots of the chemical were taken from the original test batch and transferred to the mixing area. The control test diet was prepared in the same manner as chemically-treated test diet, excluding only the test substance. A sample of each batch of feed mixed was taken and retained in the freezer until the study was complete and the analytical data deemed satisfactory. Replacement admixtures for each treatment group were prepared weekly (or at greater intervals depending on freezer stability) and stored under freezer conditions until presented to the animals the following week (or weeks). The concentration of the test substance in the feed for the females only was adjusted during the lactation period (Days 0-21) by 50%.

Details on mating procedure:

Four groups of 30 male and 30 female rats each were given 0, 30, 60, and 180 ppm of propineb in the diet seven days/week throughout the entire study. These rats were designated the P-Generation. After 10 weeks, each male was cohabited with a female in the same group, the females were allowed to litter, and wean their offspring. The offspring were designated the F1 Generation.
After weaning, F1-pups were maintained for approximately six weeks prior to initiation of the second generation. 30 male and 30 female rats from each group were selected for growth and subsequent mating to produce the F2 Generation. F2-pups were sacrificed at weaning on lactation day 21.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The concentration of Propineb in the various test diets was verified for batches intended for weeks 1, 2, 3, and at monthly intervals thereafter. Test diets intended for the first week of lactation were also analyzed. The homogeneity and stability of Propineb when mixed in the rodent feed was characterized.
Mean analytical concentrations for each dose group were 24.7, 49.1, and 153 ppm, ranging from 82-85% of the corresponding nominal concentrations of 30, 60, and 180 ppm, respectively. During lactation, the concentration of the test substance in the feed for the females was adjusted by 50%. Mean analytical concentrations for each dose group during lactation were 13.0, 25.9, and 78.9 ppm, ranging from 86-88% of the corresponding nominal concentrations of 15, 30, and 90 ppm, respectively. The AI of the test substance was not detected in the control diet. Mean recovery was 77% and ranged from 72-86% for rodent ration spiked with 14.6 ppm of Propineb and
mean recovery was 102% and ranged from 92 110% for rodent ration spiked with 180 ppm of Propineb. The mean concentrations of Propineb in the feed, sampled from three distinct layers in the mixing bowl and containing a nominal concentration of either 15- or 180- ppm, were determined to be 13.3 ppm (range 12.3-14.3 ppm; %RSD = 5.3) and 150 ppm (range 141-156 ppm; %RSD = 3.0), respectively. Based on a %RSD < to10%, Propineb was judged to be homogeneously distributed in the feed over a concentration range of 15-180 ppm. Following 7 days of room temperature storage, the analytically-determined concentration of the AI of the test substance in the 15- or 180-ppm admixture was determined to be 12.7 ppm (13.6 ppm on Day 0) and 155 ppm (146 ppm on Day 0), respectively. Following 28 days of freezer storage, the analytically-determined concentration of the AI of the test substance in the 15- and 180-ppm admixtures was determined to be 13.4 ppm (13.3 on Day 0) and 161 ppm (150 on Day 0), respectively. Propineb mixed in rodent ration was judged to be stable at room temperature for at least seven days and following freezer storage for a minimum of 28 days, over a concentration range of 15-180 ppm.

Duration of treatment / exposure:

continuously

Frequency of treatment:

Seven days/week throughout the entire study.

Details on study schedule:

- F1 parental animals not mated until 6 weeks after selected from the F1 litters.
- Selection of parents from F1 generation when pups were 21 days of age.
- Age at mating of the mated animals in the study: 6 weeks

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

180 ppm

Remarks:

High dose group, equivalent to a mean daily intake of 10.0 and 9.5 mg/kg bw/d for (P0-gen)-males and (P1-gen)- males respectively and 12.5 and 11.9 mg/kg bw/d for (P0-gen)-females and (P1-gen)-females respectively.

Doses / concentrations 2

Dose / conc.:

60 ppm

Remarks:

Mid dose group, equivalent to a mean daily intake of 3.2 and 3.0 mg/kg bw/d for (P0-gen)-males and (P1-gen)- males respectively and 4.0 and 3.6 mg/kg bw/d for (P0-gen)-females and (P1-gen)-females respectively.

Doses / concentrations 3

Dose / conc.:

30 ppm

Remarks:

Low dose group, equivalent to a mean daily intake of 1.6 mg/kg bw/d for (P0-gen)-males and (P1-gen)- males and 2.0 and 1.9 mg/kg bw/d for (P0-gen)-females and (P1-gen)-females respectively.

No. of animals per sex per dose:

30 rats/sex/group; four groups (control, 30, 60, and 180 ppm Propineb)

Control animals:

yes, concurrent no treatment

Examinations

Parental animals: Observations and examinations:

Females and males were observed (cage side) for clinical signs twice daily during the working week and at least once on weekends and holidays. Cage side observations, mortality, moribundity, behavioural changes, signs of difficult or prolonged delivery, and overt toxicity by viewing the animal in the cage were conducted. A detailed evaluation of clinical signs, and a physical examination was conducted once per week.

Body weight:
Parental animals (P and F1) body weights were recorded weekly for both males and females during the premating period. During the mating period and until sacrificed, body weights for the males were recorded once per week. During gestation, dam body weights were recorded on Days 0, 6, 13, and 20 and during lactation, on Days 0, 4, 7, 14, and 21.

Food consumption and compound intake:
Food consumption was recorded once per week for both males and females parental animals (P and F1) during the premating periods. During gestation, dam food consumption was recorded on Days 0, 6, 13, and 20 and on lactation days 0, 4, 7, 14, and 21.

Urine collection:
Prior to sacrifice, urine was collected from 10 control and 10 high dose adult males. Urine was not collected from the females. Males were individually housed throughout the day in cages fitted with urine collection trays, with food and water available. Urine was collected on ice over a one to six hour period and samples were transferred to the ultralow freezer (~ -80°C) as soon as possible after collection. After urine collection, males were transferred back to their appropriate gang cage.

Oestrous cyclicity (parental animals):

The oestrus cycle was determined by examining daily vaginal smears over a three-week period prior to mating of the P- and F1-Generation females, immediately prior to the cohabitation period. Additionally, the estrous cycle stage was determined for all females just prior to termination.

Sperm parameters (parental animals):

Sperm was collected from one testis and one epididymis for enumeration of homogenization-resistant spermatids and cauda epididymal sperm reserves, respectively at sacrifice for all P- and F1-Generation males.
In addition, an evaluation of the morphology and motility was performed on sperm sampled from the distal portion (closest to the urethra) of the vas deferens on males of the control and high dose groups of both generations. Sperm motility and counts were conducted using the Integrated Visual Operating System (IVOS,
Hamilton-Thorne Research, 1998).

Litter observations:

The PND 21 pup urine was collected overnight with up to 2 pups per sex if available from the control and high dose groups. Pups were housed throughout the day in cages fitted with urine collection trays, with food and water available. Urine was not collected on ice for the weanlings. After urine collection, pups were transferred back to their appropriate nesting cage.
The size of each litter was adjusted on lactation Day 4 to yield, as closely as possible, four males and four females per litter. If the number of male or female pups was less than four, a partial adjustment was made (e.g., three females and five males). No adjustment was made for litters of fewer than eight pups. Adjustments were made by random selection of the pups using software provided by SAS.10. Grossly abnormal pups underwent a
gross internal and external examination, and all culled pups were discarded.
The F1- and F2-pups not culled on lactation Day 4 were maintained with the dam until weaning on lactation Day 21. On lactation Day 21, a sufficient number of randomly selected F1- pups/sex/litter were maintained to produce the next generation. F1-pups not selected to become parents of the next generation were sacrificed, examined macroscopically and had organs weighed. One randomly selected pup/sex/litter for each generation had tissues collected and evaluated for any structural abnormalities or pathological changes.
Random selection of pups for selection to go to next generation and those for organ weight collection was performed using software provided by SAS

Postmortem examinations (parental animals):

All surviving parental males were sacrificed as soon as possible after the last litters were produced. Maternal animals were sacrificed following the weaning of their respective litters (lactation Day 21). F1 adult males were sacrificed after the beginning of the delivery phase for the F1-females. The animals were subjected to postmortem examinations as follows: Males were euthanized by carbon dioxide asphyxiation and a gross external examination was performed. Terminal body weights were measured and the recording of all gross pathologic alterations, weighing designated organs, and saving all gross lesions was conducted on all males.
Each dam (both P- and F1-generations) was euthanized by carbon dioxide asphyxiation and a gross external examination was performed. Terminal body weights were measured and the recording of all gross pathologic alterations, weighing designated organs, and saving all gross lesions was conducted on all females. The uterus was excised and the implantation sites, if present, were counted.
Females that were sperm positive and/or had an internal vaginal plug but did not deliver were sacrificed and necropsied after gestation Day 24. Females that were never observed as being inseminated and/or with an internal vaginal plug and did not deliver at least 24 days after the completion of the mating phase, were sacrificed and necropsied. A gross necropsy was performed on these animals as described above. In addition, patency of the cervical/uterine os in these females was examined via flushing of the uterine horns with 10% buffered formalin. The following tissues were collected (X), collected and weighed (XX), and micropathology was performed on those tissues designated with an (O) (see table 1).
Animals found moribund while on study were sacrificed and a gross necropsy performed.
Animals found dead were necropsied as soon as possible. Necropsy examinations included those parameters previously described. Pups found dead, stillborn or terminated in a moribund condition underwent a gross necropsy for possible defects and/or to determine the cause of death.

Postmortem examinations (offspring):

The F1-offspring not selected as parental animals and all F2-offspring were sacrificed at 21 days
of age. These animals were subjected to postmortem examinations (macroscopic and/or
microscopic examination) as follows.
The following tissues from 21-day weanlings were collected (X), collected and weighed (XX),
and micropathology was performed on those tissues designated with an (O) (see table 2).
Pups found dead or terminated in a moribund condition underwent a gross necropsy for possible
defects and/or cause of death.

Statistics:

Parametric data (including body weight gain and food consumption) were analyzed using a univariate Analysis of Variance (ANOVA), and if significant differences were observed, a Dunnett's Test was performed. Nonparametric data (e.g., number of estrous cycles, litter size, and number of implantation sites) were first analyzed by the Kruskal-Wallis test and then subjected to Dunn's Test if significant differences were identified. Nonparametric dichotomous data (e.g. fertility and gestation indices) were initially analyzed by the Chi-Square Test and if significance was observed between groups then by the Fisher's Exact Test with the Bonferroni adjustment. To the extent possible, the frequency of gross lesions were first examined visually, then, in the event of questionable distribution, by statistical analysis using the Chi-square and Fisher's exact tests. Sperm parameters were analyzed using ANOVA, single factor. Differences between the control and test compoundtreated
groups were considered statistically significant when p < 0.05 or p < 0.01.

Reproductive indices:

Reproductive indices: The following reproductive indices were calculated from breeding and parturition records of animals in the study:
Mating Index (%) = (# inseminated females(a) x 100)/(# of females co-housed)
Fertility Index (%) = (# of pregnant females(b) x 100)/(# of inseminated females)
Gestation Index (%) = (# of females with live pups x 100)/(# of pregnant females)
(a): Includes pregnant females not observed sperm positive or with an internal vaginal plug.
(b): Includes females which did not deliver, but had implantation sites.

Offspring viability indices:

Offspring viability indices: The following viability indices were calculated from lactation records of litters in the study:
Birth Index (%) = (total # of pups born/litter x 100)/(total # of implantation sites/litter)
Livebirth Index (%) = (# of live pups born/litter x 100)/(total # of pups/litter)
Viability Index (%) = (# of live pups/litter on day 4 (pre-culling) x 100)/(# of live pups born/litter)
Lactation Index (%) = (# of live pups/litter on day 21 x 100)/(# of live pups/litter on day 4 (post-culling))
Gestation Length = Number of whole days from day in which insemination is observed in the vaginal smear (designated Day 0 of gestation) to Lactation Day 0 (delivery of pups and entry in computer system).

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Description (incidence and severity):

There were no test substance-related mortalities or clinical observations observed during the course of this study at any dietary level tested in either generation.

Dermal irritation (if dermal study):

not examined

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

No test substance-related findings were observed on body weight and body weight gain during the study for the P0-generation males at any dietary level tested.
The P0-generation females of the 180 ppm dose group exhibited significant body weight declines for weeks 5, 9, and 10 of the premating phase. At week 10, body weights were declined 7.3%, relative to controls. A decline in body weight gain was also observed in the 180 ppm dose group (declined 32.6%, relative to controls). No test substance-related findings were observed on body weight or body weight gain during the 10-week premating phase at any other dietary level tested.
Gestation:
In the 180 ppm dose group, significant declines in body weight (mean decline Days 0-20 of 6.5%) were observed. There were no test substance-related findings observed on absolute body weight at any other dietary level tested. There were no effects observed on body weight gain.
Lactation:
There were no significant body weight declines during lactation at any dietary level tested.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

No test substance-related findings were observed on food consumption during the study for the P0-generation males and females at any dietary level tested.
Gestation:
There were no effects observed on food consumption at any dietary level tested.
Lactation:
There were no effects on food consumption considered to be test substance-related at any dietary level tested.

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Endocrine findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

There were no test substance-related microscopic findings at any dietary level tested.

Histopathological findings: neoplastic:

not examined

Other effects:

not examined

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

no effects observed

Description (incidence and severity):

There were no test item-related effects on oestrus cycle length

Reproductive function: sperm measures:

effects observed, non-treatment-related

Description (incidence and severity):

There were no effects considered to be test-substance-related on any sperm parameter evaluated at any dietary level tested for either generation.

Reproductive performance:

no effects observed

Description (incidence and severity):

Overall reproductive performance was not affected for any parameter (e.g., mating, fertility or gestation indices, days to insemination, gestation length, or the median number of implants) in either generation at any dietary level tested.

Effect levels (P0)

1

Key result

Dose descriptor:

NOAEL

Remarks:

Reproductive

Effect level:

12.5 mg/kg bw/day

Based on:

test mat.

Sex:

female

Basis for effect level:

reproductive function (oestrous cycle)

reproductive function (sperm measures)

reproductive performance

2

Key result

Dose descriptor:

NOAEL

Remarks:

Reproductive

Effect level:

10 mg/kg bw/day

Based on:

test mat.

Sex:

male

Basis for effect level:

reproductive function (oestrous cycle)

reproductive function (sperm measures)

reproductive performance

3

Key result

Dose descriptor:

NOAEL

Remarks:

systemic

Effect level:

10 mg/kg bw/day

Based on:

test mat.

Sex:

male

Basis for effect level:

clinical signs

mortality

body weight and weight gain

4

Key result

Dose descriptor:

NOAEL

Remarks:

systemic

Effect level:

4 mg/kg bw/day

Based on:

test mat.

Sex:

female

Basis for effect level:

body weight and weight gain

Target system / organ toxicity (P0)

Key result

Critical effects observed:

no

Results: P1 (second parental generation)

General toxicity (P1)

Clinical signs:

no effects observed

Description (incidence and severity):

There were no test substance-related mortalities or clinical observations observed during the course of this study at any dietary level tested in either generation.

Dermal irritation (if dermal study):

not examined

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

The P1-generation males of the 180 ppm dose group exhibited slight declines in body weight throughout premating (overall mean decline of 5.0%) with significance observed weeks 7 and 8 (mean decline for these two weeks of 6.0%, relative to controls). No effect on body weight gain was observed in this dose group.
No test substance-related findings were observed on body weight or body weight gain during the study for the P1-generation males at any other dietary level tested. Incidental body weight declines were observed in the 30 ppm dose group and is not considered to be test substance-related due to a lack of a dose response relationship. The P1-generation females of the 180 ppm dose group exhibited significant body weight declines for weeks 2 through 10 of the premating phase. At week 10, body weights were declined 11.0%, relative to controls. A decline in body weight gain was also observed in the 180 ppm dose group (declined 46.7%, relative to controls). No test substance-related findings were observed on body weight or body weight gain during the 10-week premating phase at any other dietary level tested.
Gestation:
In the 180 ppm dose group, significant declines in body weight (mean decline Days 0-20 of 9.3%) was observed. There were no test substance-related findings observed on absolute body weight at any other dietary level tested. There were no effects observed on body weight gain at any dietary level tested.
Lactation:
In the 180 ppm dose group, significant declines in body weight (mean decline Days 0-21 of 5.5%) was observed. Body weight effects were not observed at any other dietary level tested.

Food consumption and compound intake (if feeding study):

effects observed, non-treatment-related

Description (incidence and severity):

Premating: Food consumption on a g/kg/day basis was decreased during the first week of premating in the 60 and 180 ppm dose groups in males but by the second week was comparable to controls. Food consumption was unaffected by treatment at any dietary level tested in females.
Gestation:
A slight increase in food consumption on a g/kg/day basis was observed in the females of the 180 ppm dose group (significant week 13- 20). Test substance-related effects on food consumption were not observed at any other dietary level tested.

Lactation:
There were no effects on food consumption considered to be test substance-related at any dietary level tested.

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Endocrine findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

There were no terminal body weight or organ weight effects in males observed at any dietary level tested.
A decline in terminal body weight was observed in the females of the 180 ppm dose group, declined 5.4% relative to controls. Test substance-related organ weight effects were not observed at any dietary level tested.

Gross pathological findings:

no effects observed

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not examined

Other effects:

not examined

Reproductive function / performance (P1)

Reproductive function: oestrous cycle:

no effects observed

Description (incidence and severity):

There were no test article-related effects on oestrus cycle length

Reproductive function: sperm measures:

effects observed, non-treatment-related

Description (incidence and severity):

There were no effects considered to be test-substance-related on any sperm parameter evaluated at any dietary level tested for either generation. Epididymal counts for the P1-males of the 180 ppm dietary group appear low compared to the controls but are not considered to be a result of treatment with the test substance based on the following. There is a wide variability with epididymal counts, there was no reproductive consequence of this finding, and micropathology evaluation did not show any effect on the epididymis.

Reproductive performance:

no effects observed

Description (incidence and severity):

Overall reproductive performance was not affected for any parameter (e.g., mating, fertility or gestation indices, days to insemination, gestation length, or the median number of implants) in either generation at any dietary level tested.

There were no test substance-related effects observed on the mean primordial (preantral) follicles, antral follicles, or corpora luteal counts for the F1-females at any dietary level tested.

Effect levels (P1)

1

Key result

Dose descriptor:

NOAEL

Remarks:

Reproduction

Effect level:

12.5 mg/kg bw/day

Based on:

test mat.

Sex:

female

Basis for effect level:

reproductive performance

2

Key result

Dose descriptor:

NOAEL

Remarks:

Reproduction

Effect level:

9.5 mg/kg bw/day

Based on:

test mat.

Sex:

male

Basis for effect level:

reproductive performance

3

Key result

Dose descriptor:

NOAEL

Remarks:

systemic

Effect level:

3.6 mg/kg bw/day

Based on:

test mat.

Sex:

female

Basis for effect level:

body weight and weight gain

4

Key result

Dose descriptor:

NOAEL

Remarks:

systemic

Effect level:

9.5 mg/kg bw/day

Based on:

test mat.

Sex:

male

Basis for effect level:

clinical signs

mortality

body weight and weight gain

Target system / organ toxicity (P1)

Key result

Critical effects observed:

no

Results: F1 generation

General toxicity (F1)

Clinical signs:

no effects observed

Description (incidence and severity):

There were no test substance-related clinical observations observed in either generation at any dietary level tested.

Dermal irritation (if dermal study):

not examined

Mortality / viability:

no mortality observed

Description (incidence and severity):

There was no test substance-related effects observed on the viability of the pups at any dietary level tested.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Pup body weights at birth for all three treated groups were comparable to the control group.
There were no test substance-related effects on pup body weight or body weight gain observed during the lactation period at any dietary level tested.

Food consumption and compound intake (if feeding study):

not examined

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Sexual maturation:

no effects observed

Description (incidence and severity):

There were no effects observed on either vaginal patency or balanopreputial separation for the F1-pups at any dietary level tested.

Anogenital distance (AGD):

not examined

Nipple retention in male pups:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings:

no effects observed

Other effects:

not examined

Developmental neurotoxicity (F1)

Behaviour (functional findings):

not examined

Developmental immunotoxicity (F1)

Developmental immunotoxicity:

not examined

Effect levels (F1)

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

13.8 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

viability

clinical signs

body weight and weight gain

Target system / organ toxicity (F1)

Key result

Critical effects observed:

no

Results: F2 generation

General toxicity (F2)

Clinical signs:

no effects observed

Description (incidence and severity):

There were no test substance-related clinical observations observed in either generation at any dietary level tested.

Dermal irritation (if dermal study):

not examined

Mortality / viability:

no mortality observed

Description (incidence and severity):

There was no test substance-related effects observed on the viability of the pups at any dietary level tested.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Pup body weights at birth for all three treated groups were comparable to the control group.
There were no test substance-related effects on pup body weight or body weight gain observed during the lactation period at any dietary level tested.

Food consumption and compound intake (if feeding study):

not examined

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Sexual maturation:

not examined

Anogenital distance (AGD):

not examined

Nipple retention in male pups:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings:

no effects observed

Other effects:

not examined

Developmental neurotoxicity (F2)

Behaviour (functional findings):

not examined

Developmental immunotoxicity (F2)

Developmental immunotoxicity:

not examined

Effect levels (F2)

Key result

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

13.8 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

clinical signs

mortality

body weight and weight gain

Target system / organ toxicity (F2)

Key result

Critical effects observed:

no

Overall reproductive toxicity

Key result

Reproductive effects observed:

no

Any other information on results incl. tables

Tables of results are provided in the overall remarks section below.

Applicant's summary and conclusion

Conclusions:

The parental male systemic NOAEL is 180 ppm (10.0 mg Propineb/kg bw/day).
The parental female systemic NOAEL is 60 ppm (4.0 mg Propineb/kg bw/day), based on decreased body weight and/or body weight gain during premating (P and F1), gestation (P and F1) and lactation (F1) at 180 ppm (11.9 mg Propineb/kg bw/day).
The reproductive NOAEL is 180 ppm in both the males and females (10.0 mg Propineb/kg bw/day for males and 12.5 mg Propineb/kg bw/day for females) based on no test-substance- related reproductive findings observed at the highest dose tested.
The offspring NOAEL is 180 ppm (13.8 mg Propineb/kg bw/day) based on no test substance-related findings observed in the pups.

Executive summary:

In a two generation-reproduction study, Propineb was administered continuously in the feed to the Wistar rat (30 animals/dose/sex) at nominal dietary concentrations of 0, 30, 60 and 180 ppm. All test diets (including control) were available for ad libitum consumption; the homogeneity and stability of Propineb as a dietary admixture was confirmed. Body weight and food consumption determinations and detailed clinical examinations of each animal were conducted weekly throughout the study, as well as, an evaluation of multiple reproductive parameters. All animals placed on study were subject to a post-mortem examination, which included recording all gross lesions, weighing designated organs and collecting representative tissue specimens for histopathological evaluation.

 

The mean daily intake of the test substance (mg propineb/kg bw/day) throughout this two-generation reproduction study at nominal dietary concentrations of 0, 30, 60 or 180 ppm, respectively, were 1.6, 3.0-3.2 and 9.5-10.0 in the males and between 1.7 -2.1, 3.6-4.1 and 11.6-13.8 in the females. In the parent and the adult of F1 generation the main effects consisted of decreased body weight and bodyweight gain in the females after 5 week of exposure. There were no toxicological effects in the offspring. There were no reproductive effects up to highest dose level.

 

The dose level of 60 ppm (equivalent to 4.0 mg/kg bw/day) in the females and 180 ppm (equivalent to 10.0 mg/kg bw/day) in the males. The dose level of 180 ppm (equivalent to 13.8 mg/kg bw/day) was also the NOAEL for the offspring and for reproductive toxicity.