Registration Dossier

Study conducted in accordance with OECD 422 test guidance in compliance with GLP. Data included for OECD SIDS dossier. Treatment-related effects on reproduction were observed at 600 mg/kg/day. These were confined to an increase in pre-implantation losses, resulting in lower offspring numbers at this dose level. The NOAEL for reproductive toxicity was therefore considered to be 250 mg/kg/day. Read across to supporting substance, CAS No. 68442 -68 -2, by structural analogue.

A two generation reproduction toxicity study with read-across substance, CAS 68411-46-1 has been proposed in the dossier for CAS 68411 -46 -1. This dossier will be updated with the results of this study as soon as the study report is available.

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

2006

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study, which is rated as reliability 2 because it is a read-across study.

Justification for type of information:

Please refer to IUCLID Section 13 for the read-across justification.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Deviations:

not specified

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

other: Crl:CD® (SD) IGS BR

Sex:

male/female

Details on test animals or test system and environmental conditions:

Not specified

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on exposure:

The test material was administered by gavage to three groups each of ten male and ten female Sprague-Dawley Crl:CD® (SD) IGS BR strain rats, for up to fifty-four consecutive days, at dose levels of 50, 250 and 600 mg/kg/day. A control group of ten males and ten females was dosed with vehicle alone (corn oil).

Details on mating procedure:

Pairing of animals within each dose group was undertaken on a one male:one female basis on Day 15 of the study, to produce litters. The presence of sperm within the vaginal smear and/or vaginal plug in situ was taken as positive evidence of mating.

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

Not specified from the OECD SIDS documentation.

Duration of treatment / exposure:

14 days prior to mating, throughout mating and gestation and continuing through lactation day 3. Equates to:
Males: 43 days; Females: up to 54 days

Frequency of treatment:

daily

Details on study schedule:

Pairing of animals within each dose group was undertaken on a one male:one female basis on Day 15 of the study, to produce litters.

Remarks:

Doses / Concentrations:
0, 50, 250 and 600 mg/kg/day
Basis:
actual ingested

No. of animals per sex per dose:

Ten males and ten females per dose.

Control animals:

yes, concurrent vehicle

Details on study design:

Not specified

Positive control:

Not specified

Parental animals: Observations and examinations:

Clinical signs, behavioural assessments, bodyweight development, food and water consumption were monitored during the study. Haematology and blood chemistry were evaluated prior to mating on five selected males and females from each dose group.
Extensive functional observations were performed on five selected parental males from each dose group after the completion of the mating phase, and for five selected parental females rom each dose group on Day 4 post partum.

Pregnancy and parturition: The following was recorded for each female:
i) Date of mating
jj) Date and time of observed start of parturition
iii) Date and time of observed completion of parturition
iv) Duration of gestation

Oestrous cyclicity (parental animals):

Not specified

Sperm parameters (parental animals):

Not specified

Litter observations:

Daily clinical observations were performed on all surviving offspring, together with litter size and offspring weights and assessment of developmental landmarks.

Postmortem examinations (parental animals):

Males were terminated on Day 43, followed by the termination of all surviving females and offspring on Day 5 post partum. All animals were subjected to a gross necropsy examination and histopathological evaluation of selected tissues was performed.

Postmortem examinations (offspring):

Not specified

Statistics:

Not specified

Reproductive indices:

Not specified

Offspring viability indices:

Not specified

Clinical signs:

no effects observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

liver and thyroid gland

Other effects:

not examined

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

Mortality: No treatment-related deaths were detected.
Clinical Observations: No clinically observable signs of toxicity were detected.
Behavioural Assessments: No treatment-related effects were detected.
Functional Performance Tests: No treatment-related effects were detected.
Sensory reactivity Assessments: No treatment-related effects were detected.
Bodyweights: No adverse effect on bodyweight was observed for males throughout the treatment period, or for females during the maturation, gestation or lactation phases of the study.
Food Consumption: No adverse effect on dietary intake was detected for males throughout the treatment period, or for females during the maturation, gestation or lactation phases of the study.
Water Consumption: No overt intergroup differences were detected. Haematology: No treatment-related changes were detected prior to mating.
Blood Chemistry: Elevated alkaline phosphatase levels were detected for males treated with 600 mg/kg/day. Males treated with 600 and 250 mg/kg/day also showed reduced cholesterol levels. No such effects were detected for females treated with 600 or 250 mg/kg/day or for animals of either sex treated with 50 mg/kg/day.
Necropsy of Adults: No treatment-related macroscopic abnormalities were detected for the interim death female or for the remaining animals at terminal kill.
Organ Weights: Elevated liver and adrenal weights, both absolute and relative to terminal bodyweights, were detected for animals of either sex treated with 600 mg/kg/day.
Histopathology: Histopathological examination of adult tissue revealed the following treatment-related changes:
Liver: Centrilobular hepatocyte enlargement was observed for animals of either sex treated with 600 and 250 mg/g/day, with the effect extending into the female 50 mg/kg/day dose group.
Thyroid glands: Follicular cell hypertrophy was observed for males treated with 600 and probably also at 250 mg/kg/day. No such effects were detected for females at these dose levels, or for animals of either sex treated with 50 mg/kg/day.
Mating: No adverse effects on mating performance, fertility or gestation were detected.

Key result

Dose descriptor:

NOAEL

Effect level:

250 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

organ weights and organ / body weight ratios

histopathology: non-neoplastic

Clinical signs:

not examined

Mortality / viability:

mortality observed, treatment-related

Description (incidence and severity):

see Details on results (offspring)

Body weight and weight changes:

no effects observed

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings:

no effects observed

Developmental Screening:
Litter Observations: There were no clinical signs to suggest an effect of treatment.
Offspring Viability: Mean offspring weights for treated animals were comparable to controls.
Offspring Development: Offspring from the 600 mg/kg/day dose group showed less successful completion of surface righting assessments. There were no treatment-related differences in pinna unfolding.
Necropsy of Offspring: No treatment-related macroscopic abnormalities were detected for the interim death offspring or for the remaining offspring at terminal kill.

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

250 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Increase in pre-implantation losses

Reproductive effects observed:

not specified

Conclusions:

The NOAEL for reproductive toxicity was therefore considered to be 250 mg/kg/day.

Executive summary:

Study conducted to OECD test guidance in compliance with GLP. Data included for OECD SIDS dossier. Treatment-related effects on reproduction were observed at 600 mg/kg/day. These were confined to an increase in pre-implantation losses, resulting in lower offspring numbers at this dose level. The NOAEL for reproductive toxicity was therefore considered to be 250 mg/kg/day. Read across to supporting substance, CAS No. 68442 -68 -2, by structural analogue.

This substance has been supported under Environmental Protection Agency’s (EPA’s) High Production Volume (HPV) Challenge Program. The American Chemical Councils RAPA Panel, has derived a “Substituted Diphenylamines” category of chemicals for this substance, please refer to EPA reference 201-14700A located at

http://www.epa.gov/hpv/pubs/summaries/subdipha/c13378rt.pdf

Relying on several factors specified in EPA’s guidance document on “Development of Chemical Categories in the HPV Challenge Program,” in which use of chemical categories is encouraged, the chemicals constitute a chemical category on the following basis:

Structural Similarity. A key factor supporting the classification of these chemicals as a category is their structural similarity (see Figure 1). All share a common starting material; Diphenylamine (Benzenamine, N-phenyl-, CAS# 122-39-4), a common synthetic pathway, and all compounds in this category are diamines with various substitutions.

Similarity of Physicochemical Properties. The similarity of the physicochemical properties of these materials parallels their structural similarity. All are off-white to light brown solids or viscous liquids intended for use as antioxidants in finished rubber articles or as antidegradant additives that extend the useful life of heavy-duty industrial functional fluids used in high-speed, high-temperature and/or high-load applications. As a class, these amine-based antidegradant compounds are less migratory (more polymer-bound) and less staining than the Substituted p-Phenylenediamine antidegradants. The use of these materials requires that they be stable under high temperatures. Their low volatility is due to their low vapor pressure, highly viscous or solid form. The existing information for these materials indicates that they have low water solubility and high flash points.

Toxicological Similarity. Review of existing published and unpublished test data for Substituted Diphenylamines shows the aquatic and mammalian toxicity among the materials within this category are similar.

Mammalian Toxicology - Reproductive and Developmental Toxicity. Data from reproductive and developmental toxicity studies were reviewed. Sufficient data are not available to adequately represent the Substituted Diphenylamines for the purposes of the HPV Program, and additional testing is proposed, and additional testing is proposed. It is proposed to test the smallest aryl- and akyl-substituted materials.

Conclusion. Based upon the data reviewed in “Substituted Diphenylamines” category of chemicals, the physicochemical and toxicological properties of the Substituted Diphenylamine category members are similar and follow a regular pattern as a result of that structural similarity. Therefore, the definition of a chemical category has been met, and read across is considered appropriate for the category of chemical.

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

250 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Klimisch 1 study

Endpoint conclusion:

no study available

Endpoint conclusion:

no study available

This substance has been supported under Environmental Protection Agency’s (EPA’s) High Production Volume (HPV) Challenge Program. The American Chemical Councils RAPA Panel, has derived a “Substituted Diphenylamines” category of chemicals for this substance, please refer to EPA reference 201-14700A located at: http://www.epa.gov/hpv/pubs/summaries/subdipha/c13378rt.pdf

Relying on several factors specified in EPA’s guidance document on “Development of Chemical Categories in the HPV Challenge Program,” in which use of chemical categories is encouraged, the following constitute a chemical category:

Structural Similarity. A key factor supporting the classification of these chemicals as a category is their structural similarity (see Figure 1). All share a common starting material; Diphenylamine (Benzenamine, N-phenyl-, CAS# 122-39-4), a common synthetic pathway, and all compounds in this category are diamines with various substitutions.

Similarity of Physicochemical Properties. The similarity of the physicochemical properties of these materials parallels their structural similarity. All are off-white to light brown solids or viscous liquids intended for use as antioxidants in finished rubber articles or as antidegradant additives that extend the useful life of heavy-duty industrial functional fluids used in high-speed, high-temperature and/or high-load applications. As a class, these amine-based antidegradant compounds are less migratory (more polymer-bound) and less staining than the Substituted p-Phenylenediamine antidegradants. The use of these materials requires that they be stable under high temperatures. Their low volatility is due to their low vapor pressure, highly viscous or solid form. The existing information for these materials indicates that they have low water solubility and high flash points.

Toxicological Similarity. Review of existing published and unpublished test data for Substituted Diphenylamines shows the aquatic and mammalian toxicity among the materials within this category are similar.

Mammalian Toxicology - Reproductive and Developmental Toxicity. Data from reproductive and developmental toxicity studies were reviewed. Sufficient data are not available to adequately represent the Substituted Diphenylamines for the purposes of the HPV Program, and additional testing is proposed, and additional testing is proposed. It is proposed to test the smallest aryl- and akyl-substituted materials.

Conclusion. Based upon the data reviewed in “Substituted Diphenylamines” category of chemicals, the physicochemical and toxicological properties of the Substituted Diphenylamine category members are similar and follow a regular pattern as a result of that structural similarity. Therefore, the definition of a chemical category has been met, and read across is considered appropriate for the category of chemical.

In a prenatal developmental study conducted in accordance with OECD Guideline 414 with structurally similar read-across substance, CAS 36878-20-3 (at doses of 50, 150 and 500 mg/kg bw/day), no developmental toxicity was observed up to 500 mg/kg bw/day (highest dose tested). A slight decrease in body weight was noted in all treated females when compared to controls, reaching a statistical significance in females receiving 500 mg/kg bw/day (up to 7%), starting from Day 9 post coitum until the end of the study. A statistically significant decrease was also recorded in body weight gain of females of the same group on Day 9 post coitum (109%; body weight loss) and Day 12 post coitum (22%). Starting from Day 15 post coitum the mean values of body weight gain were comparable between control and high dose group. Therefore, the NOAEL for developmental toxicity was 500 mg/kg bw/day and the NOAEL for maternal toxicity was 150 mg/kg bw/day.

Reference

Endpoint:: developmental toxicity
Type of information:: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:: key study
Study period:: 2014
Reliability:: 2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:: other: Guideline study, which is rated as reliability 2 because it is a read-across study.
Justification for type of information:: Please refer to IUCLID Section 13 for the read-across justification.
Reason / purpose for cross-reference:: read-across source
Qualifier:: according to guideline
Guideline:: OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:: no
GLP compliance:: yes (incl. QA statement)
Limit test:: no
Species:: rat
Strain:: Wistar
Details on test animals or test system and environmental conditions:: TEST ANIMALS
- Source: Charles River Italia S.p.A., Calco (Lecco), Italy
- Age at study initiation: (age at delivery 10 weeks)
- Weight at study initiation: (weight range at delivery 177-196 g)
- Fasting period before study: none
- Housing: individual cages (during gestation)
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 18 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 2 °C
- Humidity (%): 55% +/- 15%
- Air changes (per hr): 15 -20
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 2014-05-20 To: 2014-06-16
Route of administration:: oral: gavage
Vehicle:: corn oil
Details on exposure:: PREPARATION OF DOSING SOLUTIONS: The formulations were prepared daily and the concentrations were calculated and expressed in terms of test item as supplied.

VEHICLE
- Justification for use and choice of vehicle: The substance is miscible in corn oil and insoluble in water.
- Concentration in vehicle: 12.5, 37.5 and 125 mg/mL
- Amount of vehicle: 4 ml/kg bw
Analytical verification of doses or concentrations:: yes
Details on analytical verification of doses or concentrations:: The proposed formulation procedure for the test item was checked in the range from 12.5 to 125 mg/mL by chemical analysis (concentration and
homogeneity) during the pre-treatment period to confirm that the method was suitable. Final results for all levels were within the acceptability limits
for concentration (90-110%). Stability after 24 hours at room temperature was verified in the range from 1 to 300 mg/mL in the validation study.
Samples of the formulations prepared on week 1 and Last Week were analysed to check the homogeneity and concentration.
Details on mating procedure:: Females were paired one to one in the home cage of the male and left overnight. Vaginal smears were taken daily in the morning from the day after pairing until a positive identification of matingwas made. The day of mating, as judged by the presence of sperm in the vaginal smear or by the presence of a copulation plug, was considered as Day 0 of gestation (or Day 0 post coitum).
Duration of treatment / exposure:: Day 6 through Day 19 post coitum
Frequency of treatment:: daily
Duration of test:: Day 6 through Day 20 post coitum
No. of animals per sex per dose:: 24
Control animals:: yes, concurrent vehicle
Details on study design:: - Dose selection rationale: Range-finding study with pregnant rats. The highest dose group of 500 mg/kg was expected to cause maternal toxicity as indicated by adverse effects on body weights and evidence on liver toxicity as indicated by clinical chemistry parameters. For details it is referred to the robust study summary of the maternal toxicity study.
- Rationale for animal assignment: Females were allocated to the groups by computerised stratified randomisation to give approximately equal initial group mean body weights
Maternal examinations:: CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations: mortality

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: Days 0, 3, 6, 9, 12, 15, 18 and 20 post coitum

FOOD CONSUMPTION : Yes
- Time schedule for examinations: Days 3, 6, 9, 12, 15, 18 and 20 post coitum starting from Day 0 post coitum

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: ovaries and uteri
Ovaries and uterine content:: The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other: gross evaluation of placentae, number of intra-uterine deaths,. Uteri or individual uterine horns without visible implantations were immersed
in a 20% solution of ammonium sulphide to reveal evidence of embryonic death at very early stages of implantation.
Fetal examinations:: - External examinations: Yes: [all per litter]
- Soft tissue examinations: Yes: [half per litter]
- Skeletal examinations: Yes: [half per litter]
Statistics:: For continuous variables the significance of the differences amongst group means was assessed by Dunnett’s test or a modified t-test, depending on the
homogeneity of data. Statistical analysis of non-continuous variables was carried out by means of the Kruskal-Wallis test and intergroup differences
between the control and treated groups assessed by a non-parametric version of the Williams test. The mean values, standard deviations and statistical
analysis were calculated from actual values in the computer without rounding off.
Indices:: Preimplantation loss
Postimplantation loss
Total implantation loss
Sex ratios
Details on maternal toxic effects:: Maternal toxic effects:yes

Details on maternal toxic effects:
A slight decrease in body weight was noted in all treated females when compared to controls, reaching a statistical significance in females receiving
500 mg/kg bw/day (up to 7%), starting from Day 9 post coitum until the end of the study.
Statistically significant decrease was also recorded in body weight gain of females of the same group on Day 9 post coitum (109%; body weight loss) and Day 12 post coitum (22%). Starting from Day 15 post coitum the mean values of body weight gain were comparable between control and high dose group.

Statistically significant decrease (up to 22%) in food consumption was observed in treated females receiving 500 mg/kg bw/day, starting from Day 9 post coitum until the end of the study.

A slight trend to decrease was observed in terminal body weight of all treated females with respect to the control. This change was about -6% in the high
dose group, without statistical significance. A statistically significant decrease in corrected body weight (up to 6%) and corrected body weight gain (up
to 50%) was noted in treated females receiving 150 and 500 mg/kg bw/day. Gravid uterus weight was similar between control and treated groups.

There were no adverse findings at the macroscopic examination at necropsy.
Dose descriptor:: NOAEL
Effect level:: 150 mg/kg bw/day (actual dose received)
Based on:: test mat.
Basis for effect level:: other: maternal toxicity
Details on embryotoxic / teratogenic effects:: Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
Litter data, mean foetal weight and sex ratio were unaffected by treatment.

A total of 15 small foetuses (foetal weight < 2.7 g) were detected: 2 out of 269 in the control group, 2 out of 214 in the low dose group, 1 out of 269 in the mid-dose group and 10 out of 254 in the high dose group. One foetus in the high dose group showed malrotation of the hindlimb, considered incidental. Of the ten small fetuses in the high dose group, 7 were from the dam which suffered most strongly from maternal toxicity as indicated by the lowest corrected body weight gain of minus 9.8g. This dam was also the only dam showing hunched posture and piloerection on gestation day 20. The higher incidence of small foetuses is therefore considered to be related to the lower maternal body weight gain.

No relevant findings that could be considered treatment-related were observed at visceral examination of foetuses in the treated groups, compared to controls.
The alterations recorded at skeletal examinations of foetuses were noted both in control and treated groups with a similar incidence.
Dose descriptor:: NOAEL
Effect level:: >= 500 mg/kg bw/day (actual dose received)
Based on:: test mat.
Basis for effect level:: other: teratogenicity
Dose descriptor:: NOAEL
Effect level:: >= 500 mg/kg bw/day (actual dose received)
Based on:: test mat.
Basis for effect level:: other: embryotoxicity
Abnormalities:: not specified
Developmental effects observed:: not specified
Conclusions:: The substance is not teratogenic and not embryotoxic in rats. It causes maternal toxicity at a dose level of 500 mg/kg bw.

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

150 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Klimisch 1 study

Endpoint conclusion:

no study available

Endpoint conclusion:

no study available

This substance has been supported under Environmental Protection Agency’s (EPA’s) High Production Volume (HPV) Challenge Program. The American Chemical Councils RAPA Panel, has derived a “Substituted Diphenylamines” category of chemicals for this substance, please refer to EPA reference 201-14700A located at http://www.epa.gov/hpv/pubs/summaries/subdipha/c13378rt.pdf

Relying on several factors specified in EPA’s guidance document on “Development of Chemical Categories in the HPV Challenge Program,” in which use of chemical categories is encouraged, the following closely related chemicals constitute a chemical category:

Structural Similarity. A key factor supporting the classification of these chemicals as a category is their structural similarity (see Figure 1). All share a common starting material; Diphenylamine (Benzenamine, N-phenyl-, CAS# 122-39-4), a common synthetic pathway, and all compounds in this category are diamines with various substitutions.

Similarity of Physicochemical Properties. The similarity of the physicochemical properties of these materials parallels their structural similarity. All are off-white to light brown solids or viscous liquids intended for use as antioxidants in finished rubber articles or as antidegradant additives that extend the useful life of heavy-duty industrial functional fluids used in high-speed, high-temperature and/or high-load applications. As a class, these amine-based antidegradant compounds are less migratory (more polymer-bound) and less staining than the Substituted p-Phenylenediamine antidegradants. The use of these materials requires that they be stable under high temperatures. Their low volatility is due to their low vapor pressure, highly viscous or solid form. The existing information for these materials indicates that they have low water solubility and high flash points.

Toxicological Similarity. Review of existing published and unpublished test data for Substituted Diphenylamines shows the aquatic and mammalian toxicity among the materials within this category are similar.

Mammalian Toxicology - Reproductive and Developmental Toxicity. Data from reproductive and developmental toxicity studies were reviewed. Sufficient data are not available to adequately represent the Substituted Diphenylamines for the purposes of the HPV Program, and additional testing is proposed, and additional testing is proposed. It is proposed to test the smallest aryl- and akyl-substituted materials.

Conclusion. Based upon the data reviewed in “Substituted Diphenylamines” category of chemicals, the physicochemical and toxicological properties of the Substituted Diphenylamine category members are similar and follow a regular pattern as a result of that structural similarity. Therefore, the definition of a chemical category has been met, and read across is considered appropriate for the category of chemical.

Justification for selection of Effect on developmental toxicity: via oral route:
Available study data for the Substituted Diphenylamines group

The above studies have been ranked reliability 1 according to the Klimisch et al system. This ranking was deemed appropriate because the studies were conducted to GLP and in compliance with agreed protocols. Sufficient dose ranges and numbers are detailed; hence it is appropriate for use based on reliability and animal welfare grounds. As the effects are considered adaptive rather than toxicological, no classification is proposed.

The above results triggered no classification under the CLP Regulation (EC No 1272/2008).

Group		Terminal body weight (g)	Gravid uterus weight (g)	Body weight at necropsy minus gravid uterus weight (corrected body weight) (g)	Body weight at necropsy minus gravid uterus weight, minus body weight on GD6 (corrected body weight gain) (g)
1 (control)	Mean	336.38	66.40	269.98	29.87
	SD	15.84	8.80	14.79	7.67
	(n)	23	23	23	23
2	Me	323.9	60.42	263.5	29.45
	SD	23	18	18	16
	(n)	20	20	20	20
3	Me	323.8	66.20	257.6*	20.5*
	SD	28	12	18	9.
	(n)	23	23	23	23
4	Me	315.9	62.53	253.3*	14.83*
	SD	24	14	23	16
	(n)	22	22	22	22

Group		gestation day 3	gestation day 6	gestation day 9	gestation day 12	gestation day 15	gestation day 18	gestation day 20
1 (control)	n	23	23	23	23	23	23	23
	Mean	18.13	20.66	18.72	20.99	21.78	23.68	23.06
	SD	2.37	2.54	2.05	1.98	2.10	2.20	1.99
2	n	20	20	20	20	20	20	20
	Mean	17.99	20.97	18.32	20.64	21.04	23.43	22.83
	SD	2.3	3.03	2.46	2.19	2.68	2.99	4.33
3	n	23	23	23	23	23	23	23
	Mean	18.7	20.23	17.36	19.28	20.90	22.32	21.56
	SD	1.99	2.53	2.59	2.90	2.40	2.91	3.14
4	n	22	22.00	22	22.00	22.00	22.00	22.00
	Mean	18.24	20.97	15.33**	17.1**	19.01**	20.12**	18.02**
	SD	2.62	2.55	1.98	2.76	2.68	3.02	2.96

Group	1 (control)	2	3	4
Initial group size (n)	24	24	24	24
Not pregnant (n)	1	4	1	2
Unilateral implantation (n)	0	1	0	0
With live foetuses at gestation Day 20 (n)	23	20	23	22

		Corpus lutea	Implantations	Early Uterine Deaths	Late Uterine Deaths	Total Uterine Deaths	Viable Young (total)	Viable males	Viable females	% Males	Preimplantation loss (%)	Postimplantation loss	Total implantation loss(%)	Litter weight (g)	Mean fetal weight (g)
1	Mean	12.87	12.26	0.52	0.04	0.57	11.7	6	5.7	50.96	4.68	4.15	8.52	43.1	3.69
	SD	1.63	1.63	1.47	0.21	1.5	1.77	2.24	2.05	17.27	5.13	10.27	11.78	6.71	0.34
	(n)	23	23	23	23	23	23	23	23	23	23	23	23	23	23
2	Mean	12.26	11.47	0.21	0.05	0.26	11.21	5.67	5.84	49.82	6.56	4.12	10.05	39.88	3.63
	SD	2.58	2.44	0.54	0.23	0.56	2.84	2.22	2.29	14.18	6.13	11.67	13.68	9.32	0.4
	(n)	19	19	19	19	19	19	18	19	18	19	19	19	19	19
3	Mean	13.09	12.22	0.52	0	0.52	11.7	5.78	5.91	449.19	6.28	4.73	10.86	42.41	3.65
	SD	2.7	2.35	0.73	0	0.73	2.55	1.93	1.98	12.79	6.68	6.74	7.39	8.85	0.28
	(n)	23	23	23	23	23	23	23	23	23	23	23	23	23	23
4	Mean	12.27	11.73	0.14	0	0.14	11.59	5.36	6.23	46.95	6.11	1.18	7.23	40.11	3.51
	SD	2.66	3.06	0.35	0	0.35	3.08	2.06	2.25	12.97	10.6	3.07	10.96	9.86	0.39
	(n)	22	22	22	22	22	22	22	22	22	22	22	22	22	22