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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
2019-
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Deviations:
yes
Remarks:
ten weeks premating administration period, estrous cycle determination performed at the end of the premating administration period (in a three weeks period before the start of mating)
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid: viscous
Details on test material:
100 g/100 g (UVCB substance)


Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material:
BASF and 0016046440
- Expiration date of the lot/batch:
07 Jan 2020

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material:
ambient (room temperatures)
- Stability under test conditions: guaranteed by the sponsor
- Solubility and stability of the test substance in the solvent/vehicle:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing:

- Preliminary purification step (if any):
- Final dilution of a dissolved solid, stock liquid or gel:
- Final preparation of a solid:

FORM AS APPLIED IN THE TEST: test substance addition to the diet

Test animals

Species:
rat
Strain:
Wistar
Remarks:
Crl:Wl(Han)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 35±1 d
- Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x g
- Housing: group housed - up to 5 animals/cage (pretreatment), 2 animals/cage (premating), 2 male animals/cage (mating and postmating)
- Polysulfonate cages Typ 2000P (H-Temp)
- Diet: Mouse and rat maintenance diet "GLP", Granovit AG, Kaiseraugst, Switzerland; ad libitum
- Water: Drinking water ad libitum
- Acclimation period: 7 d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 45 - 65
- Air changes (per hr): 15
- Photoperiod (hrs dar k / hrs light): 12/12

IN-LIFE DATES: From: 2019-05-21 To: 2019-09-25 (2019-10-10 recovery females)

Administration / exposure

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: 14 days
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 (GD 0) of pregnancy
- After successful mating each pregnant female was caged: individually
Details on analytical verification of doses or concentrations:
will be carried out as a separate study at the test facility Competence Center Analytics of BASF SE, 67056 Ludwigshafen, Germany
Frequency of treatment:
daily for 7 days a week
Doses / concentrationsopen allclose all
Dose / conc.:
500 ppm
Remarks:
dose applied to animals during the premating and mating phase (males and females), postmating (males) and gestation phase and after weaning (females)
Dose / conc.:
1 500 ppm
Remarks:
dose applied to animals during the premating and mating phase (males and females), postmating (males) and gestation phase and after weaning (females)
Dose / conc.:
5 000 ppm
Remarks:
dose applied to animals during the premating and mating phase (males and females), postmating (males) and gestation phase and after weaning (females);
dose was also applied to recovery animals
Dose / conc.:
250 ppm
Remarks:
dose applied to F0 females during lactation phase to maintain dams at desired target dose of the test substance: adjustment to historical body weight and food consumption data
Dose / conc.:
750 ppm
Remarks:
dose applied to F0 females during lactation phase to maintain dams at desired target dose of the test substance: adjustment to historical body weight and food consumption data
Dose / conc.:
2 500 ppm
Remarks:
dose applied to F0 females during lactation phase to maintain dams at desired target dose of the test substance: adjustment to historical body weight and food consumption data
No. of animals per sex per dose:
10
Control animals:
yes
Details on study design:
- Dose selection rationale: requested by the sponsor
- Rationale for animal assignment: rats will be randomized according to their weight and allocated to the dose groups
- Fasting period before blood sampling for clinical biochemistry: 16-20 hours

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule for moribund and dead animals: twice daily (Mon-Fri), once daily (Sat, Sun and public holidays)
- Time schedule for clinical signs: at least once daily, if signs occur: several times daily

DETAILED CLINICAL OBSERVATIONS: Yes / No / No data
- Time schedule:

BODY WEIGHT: Yes
- Time schedule for examinations: once a week for animals in the main group and the recovery group
- not determined in females without positive evidence of sperm during mating and gestation periods and in the females without litter during lactation period
- during mating, females will be weighed on the day of positive evidence of sperm (GD 0) and on GD 7, 14 and 20
- females with litter will be weighed on the day after parturition (PND1) and on PND 4, 7, 10 and 13

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption will be determined once a week for the male and female parental animals of main groups as well as the males and females of recovery groups.
- not be determined during mating, in the females without positive evidence of sperm during mating and gestation periods and in the females without litter during lactation period
- females with evidence of sperm: food consumption will be determined for GD 0-7, 7-14 and 14-20
- females which gave birth to a litter: food consumption will be determined for PND 1-4, 4-7, 7-10 and 10-13

OTHER:
Oestrous cyclicity (parental animals):
- in all parental females in the premating phase: estrous cycle length and normality will be evaluated by preparing vaginal smears during a minimum of 3 weeks prior to mating and throughout cohabitation until there is evidence of sperm in the vaginal smear
- in all females of the recovery groups: estrous cycle length and normality will be evaluated in the 2 weeks of the recovery period
- in all parental females: estrous status will be determined on the day of scheduled sacrifice
Sperm parameters (parental animals):
Parameters examined in F0 male parental generations and males of the recovery group:
- after organ weight determination, parameters determined in the right testis or right epididymis:
- Cauda epididymis sperm motility
- Sperm morphology
- Spermatid head count in the testis
- Sperm head count in the cauda epididymis
Parameters exmined for the control and highest dose group:
- Sperm morphology and sperm head count (cauda epididymis and testis)
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes
- If yes, maximum of 8 pups/litter (4/sex/litter as nearly as possible); excess pups were killed and discarded.

Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals after the end of the administration period the last litters in each generation were produced.
- Female animals: All surviving animals after the last litter of each generation was weaned (PND 14).
- Recovery animals: All surviving animals will be maintained for at least two weeks (recovery period) longer compared to the respective main groups.

GROSS NECROPSY
- Gross necropsy performed for all animals in the main and the recovery group
- consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera
- special attention being given to reproductive organs

HISTOPATHOLOGY / ORGAN WEIGHTS
Organs/Tissues weighed for...:
- all animals (main test and recovery group): anesthetized animals (final body weight), Epididymides, Ovaries, Prostate (ventral and dorsolateral part together, fixed), Seminal vesicles with coagulating glands (fixed), Testes, Thyroid glands (with parathyroid glands) (fixed), Uterus with cervix
- 5 animals/sex/test group (main test and recovery group): Adrenal glands (fixed), Brain, Heart, Kidneys, Liver, Spleen, Thymus (fixed)
Organs/Tissues fixed (4% netrual buffered formaldehyde) for...:
- all parental animals (main test and recovery group): All gross lesions, Adrenal glands, Aorta, Bone marrow (femur), Brain, Cecum, Cervix, Coagulating glands, Colon, Duodenum, Esophagus, Extraorbital lacrimal glands, Epididymides, left (modified Davidson's solution), Eyes with optic nerve (modified Davidson's solution), Femur with knee joint, Heart, Ileum, Jejunum (with Peyer's patches), Kidneys, Larynx
Postmortem examinations (offspring):
SACRIFICE
- One selected male and one female pup/litter were sacrificed at 13 days of age (PND 13) for blood sampling and thyroid/parathyroid glands fixation.
- All F1 offspring
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:
Statistics:
Beside means and standard deviations, details statistics are given in table 1.

Results and discussion

Applicant's summary and conclusion