2,2'-(octylimino)bisethanol

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

FEBRUARY 2021 to JUNE 2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2021

Report date:

2021

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EPA OPP 83-3 (Prenatal Developmental Toxicity Study)

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Version / remarks:

Adopted 25 June, 2018

GLP compliance:

yes (incl. QA statement)

Remarks:

This study was audited by the Quality Assurance, Vimta Labs Ltd., in compliance with the Organisation for Economic Co-operation and Development (OECD) Principles of Good Laboratory Practice (GLP) ENV/MC/CHEM (98) 17, and the findings were reported to the

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

2,2'-(octylimino)bisethanol

EC Number:

239-555-0

EC Name:

2,2'-(octylimino)bisethanol

Cas Number:

15520-05-5

Molecular formula:

C12H27NO2

IUPAC Name:

2-[(2-hydroxyethyl)(octyl)amino]ethan-1-ol

Specific details on test material used for the study:

Name Genamin 3920
Chemical Name 2,2’-(Octylimino)bisethanol
Manufacturer’s / Sponsor’s Name and Address Clariant Produkte (Deutschland) GmbH, Corporate Product Stewarship on behalf of BU ICS
Bruningstr. 50, 65926 Frankfurt, Germany
Molecular Formula C12H27NO2
CAS-No. 15520-05-5
Molecular weight 217.35 g/mol
Physical Appearance Transparent liquid
Storage Conditions Ambient (Room Temperature 20-25 °C)

Test animals

Species:

rat

Strain:

Wistar

Details on test animals or test system and environmental conditions:

Species : Rat (Rattus norvegicus)
Strain : Wistar
Age : 10 - 12 weeks at start of treatment
Sex : Pregnant females
Body Weight : 217.23 – 341.45 g at start of treatment
Number of Animals : 96 pregnant females (dams)
Animals were maintained under the following environmental conditions:
Temperature : 20.6 – 24.3°C
Relative humidity : 38 – 67%
Light/dark cycle (photoperiod) : 12-hour light and 12-hour dark cycle

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The concentration of Genamin 3920 in the dose formulations were in the range between 97.5 – 105.1 which were within the acceptance criteria (85% - 115%). No degraded impurity was observed during the 48h stability test period. The results revealed homogenous distribution of Genamin 3920 in the dose formulation.

Duration of treatment / exposure:

Dose formulations of vehicle and test item were administered to respective groups by oral route once daily from GD 5 to GD 19 of presumed gestation. Dose was administered using a calibrated disposable syringe attached to a 16 and18-gauge stainless steel ball tipped oral gavage needle. Individual animal dose volumes were calculated based on their most recent body weight. Homogeneity of the dose formulations during sampling and dosing were maintained by constant stirring using a magnetic stirrer.

Frequency of treatment:

Once daily from Gestation Day 5 to Gestation Dat 19

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

75 mg/kg bw/day

Doses / concentrations 2

Dose / conc.:

150 mg/kg bw/day

Doses / concentrations 3

Dose / conc.:

300 mg/kg bw/day

Remarks:

Animals were initially dosed at 350 mg/kg. As overt toxicity was observed, dose wad reduced to 300 mg/kg/day.

No. of animals per sex per dose:

24 presumed pregnant females

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: In a dose range finding study (Vimta Study No. VLL/1020/NG/T108) pregnant dams were treated at dose levels of 50, 150 and 300 mg/kg b.w./day. In this study, a slight but not statistically significant decrease in body weight and feed consumption was observed in the highest dose (300 mg/kg). In another repeated dose 90-day oral toxicity study (Vimta Study No. VLL/1020/G/T107), no treatment related clinical signs and mortality occurred at the highest dose of 350 mg/kg b.w./day, after 75 days of treatment. The doses for the current study are selected taking into consideration, the outcome of the above-mentioned studies. In a subacute repeated dose oral toxicity study (according to OECD 407) (Study No. 134233) male and female Wistar rats were treated with 20, 100, and 500 mg/kg b.w./day of the test item for 28 days. Since at a daily oral dose of 500 mg/kg b.w./day, 5/10 female animals died during the treatment period, 350 mg/kg per day was selected as the highest tolerable dose for pregnant dams.
- Rationale for animal assignment (if not random): Pregnant females received on a particular day were randomly allocated to 4 groups (G1- G4; 1 vehicle control group and 3 test groups) ensuring minimum differences in mean body weight between the groups. This procedure was continued till each group gets the required number of females (dams).

Examinations

Maternal examinations:

The following maternal data was recorded.
• Pregnancy status
• Uterine weight
• No. of corpora lutea
• Total/viable/dead foetuses
• No. of implantation sites
• Total/early/late resorptions
Based on the uterine observations the following parameters were calculated as per
Formulas mentioned in Appendix.10
• Adjusted Maternal Weight
• Relative Uterus Weight
• Pre and Post Implantation Loss
• Percentage live and dead implants
• Percentage live offspring

Ovaries and uterine content:

Following termination, as soon as possible after death, the uteri were removed (Caesarean section) and the pregnancy status of the animals ascertained. Uteri that appear non-gravid was examined with ammonium sulphide staining to confirm the non-pregnant status. The gravid uterus along with cervix was weighed immediately.

Blood sampling:

On gestation day 20, blood (approximately 2 ml) was collected from all surviving dams through retro-orbital plexus in tubes (without anticoagulant) under mild isoflurane anaesthesia.

Fetal examinations:

Foetal Examination: All the foetuses were removed sequentially and were identified by serial numbers as per SOP.
The following litter data was recorded:
• Sex identification
• Body Weight
• Anogenital distance
• Individual foetus and placenta weight (g) (placenta was discorded)
All the foetuses were examined for external malformations.

Visceral and Head Razor Examination: Approximately half of the live foetuses per litter were selected. A detailed soft tissue examination was performed as per in-house SOP, which included observation of all the organs and structures of the head, neck, thorax and abdomen. Reproductive tract was examined for signs of altered development. External foetal sex (as determined by gross examination) was compared internally (gonadal observations). The heads of foetuses were decapitated and preserved in Bouin’s fixative for head razor examination.

Skeletal Examination: Foetuses selected for skeletal examination were eviscerated and fixed in alcohol. Detailed examinations of the skeleton (bone + cartilage) were performed after staining with alizarin red S and alcian blue. Examination included observation of all the bone structures and cartilage of the head, spine, rib cage, pectoral, pelvis and limbs.

Statistics:

The data was analysed using Graphpad Prism, version 4.00. Statistical comparisons were carried out between treatment and control groups using parametric (one-way analysis of variance (ANOVA), Dunnett's t-Test) or non-parametric (Kruskal-Wallis test, Mann Whitney’s Test) test procedures. The choice of parametric or non-parametric test was based on whether the groups satisfy the homogeneity of variance as evaluated by Bartlett’s test. Statistical significance was evaluated at p≤0.05 and/or p≤0.01. All quantitative data were summarized and expressed as Mean ± SD.

Historical control data:

Historical data provided in the Study Report to allow comparison to control data.

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Three dams were found dead between GD 9-15 with clinical signs/symptoms at 350 mg/kg body weight/day. Animals treated at 350 mg/kg/day displayed salivation, hyperactivity, aggression until reduction of dose to 300 mg/kg/day.

Mortality:

mortality observed, treatment-related

Description (incidence):

Three dams were found dead between GD 9-15

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Maternal body weight was significantly reduced from GD 8-20 in dams treated with Genamin 3920 at the dose of 350/300 mg/kg. All dams belonging to the control, low and mid dose treated groups displayed normal body weight gain throughout the study period. Adjusted maternal weight (after exclusion of uterine weight) of treated dams derived on GD 20 was significantly reduced in the 350/300 mg/kg group.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Feed consumption was significantly reduced from GD 8-20 in dams treated with Genamin 3920 at the dose of 350/300 mg/kg.

Endocrine findings:

no effects observed

Description (incidence and severity):

No changes in T3, T4 and TSH attributable to treatment were observed. Further, no histopathological changes in thyroid/parathyroid was observed in any of the treated groups as compared to the control.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

In the found dead dams treated at 350 mg/kg/day, gross pathological evaluations revealed red discoloration in lung and mottled liver

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Histopathological observations of the found dead animals revealed kidneys with necrosis, bilateral, multifocal (3+) in 2 dams and kidneys with necrosis, bilateral, multifocal (2+), hyperplasia with squamous cell, non-glandular stomach, diffuse (3+) in 1 dam in the high dose treated group of 350 mg/kg body weight/day.

Maternal developmental toxicity

Number of abortions:

no effects observed

Pre- and post-implantation loss:

no effects observed

Total litter losses by resorption:

no effects observed

Early or late resorptions:

no effects observed

Dead fetuses:

no effects observed

Changes in pregnancy duration:

no effects observed

Changes in number of pregnant:

no effects observed

Other effects:

no effects observed

Details on maternal toxic effects:

Surviving dams of the high dose group treated at 350/300 mg/kg/day revealed significant decrease in the gravid uterine weights as compared to the control group.
No other abnormalities were observed in the evaluated maternal parameters viz., placental weight, number of corpora lutea, implantations, resorptions and percent implantation loss in treated (75, 150 and 350/300 mg/kg/day) dams as compared to control group.

Effect levels (maternal animals)

Key result

Dose descriptor:

NOAEL

Effect level:

>= 150 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

body weight and weight gain

clinical signs

food consumption and compound intake

gross pathology

histopathology: non-neoplastic

mortality

necropsy findings

Maternal abnormalities

Key result

Abnormalities:

effects observed, treatment-related

Localisation:

other: Body weight and gravid uterine weight

Results (fetuses)

Fetal body weight changes:

no effects observed

Reduction in number of live offspring:

no effects observed

Changes in litter size and weights:

no effects observed

Anogenital distance of all rodent fetuses:

no effects observed

External malformations:

no effects observed

Skeletal malformations:

no effects observed

Visceral malformations:

no effects observed

Details on embryotoxic / teratogenic effects:

There were no significant changes in the mean number of male and female foetuses and total number of foetuses in any of the treated groups (75, 150 and 350/300 mg/kg/day) as compared to the control group. Foetal weight was comparable between treated and control groups. No malformations related to treatment were detected during external examination of the pups. Few incidences of haematoma in right pinna and lower mandible (jaw) observed in Genamin 3920 treated dams at 75 mg/kg and 350/300 mg/kg (0.3% and 0.4%), were within normal historical ranges and hence considered as common spontaneous lesions.

No malformations/abnormalities were detected in any of the treated and control group pups during the visceral and head razor examinations except for a few incidental findings in control and treated groups (like haemorrhages in lungs, adrenals, kidney and in lateral ventricle of brain). The incidence and severity of the observations were within the normal historical control ranges.

Foetal skeletal examination did not reveal any treatment related malformations/ abnormalities in any of the treated and control groups.

Effect levels (fetuses)

Key result

Dose descriptor:

NOAEL

Effect level:

>= 300 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Remarks on result:

not determinable due to absence of adverse toxic effects

Fetal abnormalities

Key result

Abnormalities:

no effects observed

Overall developmental toxicity

Key result

Developmental effects observed:

no

Any other information on results incl. tables

Summary of Maternal and Litter Data

Parameters	Group & Dose (mg/kg b.wt/day)
	G1 (0)	G2 (75)	G3 (150)	G4 (350/300)
No. of rats / group	24	24	24	24
Duration of treatment (GD5-GD19)	15 days	15 days	15 days	15 days
Caesarean section carried out on	GD 20	GD 20	GD 20	GD 20
No. of rats found non-pregnant / group	2	1	1	2
No. of rats found pregnant / group	22	23	23	20
No. of rats found dead / group	0	0	0	3
No. of litters examined	22	23	23	20
No. of resorptions (early)	16	28	11	24
No. of resorptions (late)	4	7	7	5
Total no. of foetuses	340	331	360	281
Mean litter size	15	14	15	14
No. of foetuses evaluated
a.  External examination	340	331	360	281

Summary of Clinical Signs

Group & Dose (mg/kg)	Clinical Signs	No. of Dams showing Clinical Signs
		Gestational Day
		0	1	2	3	4	5	6	7	8	9	10
G1 (0)	Normal	24	24	24	24	24	24	24	24	24	24	24
G2 (75)	Normal	24	24	24	24	24	24	24	24	24	24	24
G3 (150)	Normal	24	24	24	24	24	24	24	24	24	24	24
G4 (350/300)	Normal	24	24	24	24	24	21	10	4	7	8	9
	Burrowing Behaviour	0	0	0	0	0	4	14	20	17	14	13
	Salivation	0	0	0	0	0	0	0	2	8	14	13
	Hyperactivity	0	0	0	0	0	0	0	0	0	1	5
	Aggression	0	0	0	0	0	0	0	0	0	0	1
	Found Dead	0	0	0	0	0	0	0	0	0	2	0

Group & Dose (mg/kg)	Clinical Signs	No. of Animals showing Clinical Signs
		Gestational Day
		11	12	13	14	15	16	17	18	19	20
G1 (0)	Normal	24	24	24	24	24	24	24	24	24	24
G2 (75)	Normal	24	24	24	24	24	24	24	24	24	24
G3 (150)	Normal	24	24	24	24	24	24	24	24	24	24
G4 (350/300)	Normal	9	11	11	11	13	14	17	21	21	21
	Burrowing Behaviour	11	11	11	9	7	4	1	0	0	0
	Salivation	11	11	11	9	7	4	1	0	0	0
	Hyperactivity	7	5	5	6	5	4	2	0	0	0
	Aggression	4	5	5	6	4	6	2	0	0	0
	Found Dead	0	0	0	0	1	0	0	0	0	0

Group & Dose (mg/kg)	Clinical Signs	No. of Animals showing Clinical Signs
		Gestational Day
		11	12	13	14	15	16	17	18	19	20
G1 (0)	Normal	24	24	24	24	24	24	24	24	24	24
G2 (75)	Normal	24	24	24	24	24	24	24	24	24	24
G3 (150)	Normal	24	24	24	24	24	24	24	24	24	24
G4 (350/300)	Normal	9	11	11	11	13	14	17	21	21	21
	Burrowing Behaviour	11	11	11	9	7	4	1	0	0	0
	Salivation	11	11	11	9	7	4	1	0	0	0
	Hyperactivity	7	5	5	6	5	4	2	0	0	0
	Aggression	4	5	5	6	4	6	2	0	0	0
	Found Dead	0	0	0	0	1	0	0	0	0

 

Summary of Maternal Body Weight (g)

Gestational Day	Group & Dose (mg/kg b.w.)
	G1 (0)			G2 (75)			G3 (150)			G4 (350/300)
	Mean	SD	N	Mean	SD	N	Mean	SD	N	Mean	SD	N
0	241.37	22.98	22	238.89	19.23	23	246.56	18.79	23	235.99	18.71	20
3	256.12	24.73	22	252.38	18.78	23	259.14	19.27	23	249.99	17.99	20
5	265.03	27.64	22	261.13	18.40	23	269.80	18.23	23	258.21	19.34	20
8	276.40	28.73	22	272.68	20.04	23	277.70	19.55	23	250.62↓↓	25.81	20
11	292.89	29.27	22	287.61	20.45	23	293.80	18.99	23	266.99↓↓	24.79	20
14	310.27	28.94	22	303.76	19.82	23	310.52	19.11	23	283.36↓↓	21.74	20
17	345.04	31.21	22	336.66	20.31	23	344.82	20.18	23	312.51↓↓	26.65	20
20	393.18	34.44	22	385.65	22.17	23	394.24	22.63	23	355.42↓↓	32.97	20
20*	301.95	29.49	22	299.77	21.08	23	302.22	19.14	23	276.85↓↓	25.00	20

Key: N = Number of Dams, SD = Standard Deviation, * = Adjusted Maternal body Weight, ↓↓ = significantly high at P ≤ 0.01

 

Summary of Dam Body Weight Gain (g)

Gestational Day	Group & Dose (mg/kg b.w.)
	G1 (0)			G2 (75)			G3 (150)			G4 (350/300)
	Mean	SD	N	Mean	SD	N	Mean	SD	N	Mean	SD	N
0-3	6.12	1.98	22	5.70	1.31	23	5.17	3.22	23	6.00	1.93	20
3-5	3.42	1.33	22	3.51	1.77	23	4.20	3.05	23	3.28	1.40	20
5-8	4.33	2.36	22	4.41	1.58	23	2.92	1.67	23	-3.03↓↓	5.36	20
8-11	6.05	3.04	22	5.51	2.24	23	5.85	1.72	23	6.72↓↓	4.37	20
11-14	6.02	2.48	22	5.67	1.51	23	5.73	1.65	23	6.33↓↓	3.73	20
14-17	11.25	1.85	22	10.91	2.92	23	11.08	2.00	23	10.27↓↓	3.53	20
17-20	14.01	2.80	22	14.61	3.08	23	14.35	1.86	23	13.76↓↓	5.04	20
0-5	9.75	2.36	22	9.41	1.90	23	9.51	2.08	23	9.47	2.31	20
5-20	48.73	6.58	22	47.95	6.72	23	46.29	4.94	23	37.93↓↓	11.79	20
0-20	63.19	7.20	22	61.92	8.77	23	60.24	7.21	23	51.02↓↓	13.45	20

Key: N = Number of Dams, SD = Standard Deviation, ↓↓ = significantly high at P ≤ 0.01, Note: Body weight gain considered only for pregnant dams

 

Summary of T3, T4, TSH Values

Parameters	Group & Dose (mg/kg b.w.)
	G1 (0)			G2 (75)			G3 (150)			G4 (350/300)
	Mean	SD	N	Mean	SD	N	Mean	SD	N	Mean	SD	N
T3	25.90	6.02	22	29.99	7.71	23	31.34	7.23	23	28.97	9.43	20
T4	1.08	0.24	22	1.12	0.25	23	1.08	0.21	23	1.04	0.33	20
TSH	2.21	0.91	22	2.61	1.25	23	2.36	1.16	23	2.60	1.37	20

Key: N = Number of Dams, SD = Standard Deviation

Note: Values of pregnant dams alone were considered.

 

Summary of Dam Examination and Derived Uterine Parameters

Parameters	Group & Dose (mg/kg b.w.)
	G1 (0)			G2 (75)			G3 (150)			G4 (350/300)
	Mean	SD	N	Mean	SD	N	Mean	SD	N	Mean	SD	N
Uterine Weight (g)	91.24	10.17	22	85.88	14.41	23	92.02	12.11	23	78.57↓↓	16.71	20
Relative Uterine Weight (%)	23.24	2.12	22	22.26	3.54	23	22.38	5.96	23	21.99	3.94	20
Placental Weight (g)	0.55	0.08	22	0.54	0.07	23	0.53	0.08	23	0.51	0.08	20
Total No. of Corpora Lutea	18.45	3.33	22	17.52	2.17	23	18.09	2.43	23	17.40	1.88	20
Total No. of Implantations	16.36	1.50	22	15.91	1.76	23	16.43	1.65	23	15.50	2.26	20
Total No. of live Implantations	15.45	1.79	22	14.39	2.59	23	15.65	2.27	23	14.05	2.86	20
Total No. of dead Implantations	0.00	0.00	22	0.00	0.00	23	0.00	0.00	23	0.00	0.00	20
Early Resorptions (No.)	0.73	0.88	22	1.22	1.78	23	0.48	0.67	23	1.20	1.99	20
Late Resorptions (No.)	0.18	0.50	22	0.30	0.70	23	0.30	0.70	23	0.25	0.55	20
Total Resorptions (No.)	0.91	0.97	22	1.52	2.29	23	0.78	1.09	23	1.45	2.14	20
Pre-implantation Loss (%)	9.36	12.85	22	8.50	10.41	23	8.38	8.99	23	10.74	10.27	20
Post Implantation Loss (%)	5.60	6.24	22	9.29	14.25	23	5.04	7.49	23	9.32	13.84	20
Live Foetus (%)	94.40	6.24	22	90.71	14.25	23	94.96	7.49	23	90.68	13.84	20
Dead Foetus (%)	0.00	0.00	22	0.00	0.00	23	0.00	0.00	23	0.00	0.00	20

Key: N = Number of Dams, SD = Standard Deviation, ↓↓ = significantly high at P ≤ 0.01

 

Summary of Litter data

Parameters		Group & Dose (mg/kg b.w.)
		G1 (0)	G2 (75)	G3 (150)	G4 (350/300)
Total No. of litters		22	23	23	20
Total No. of foetuses		340	331	360	281
Mean litter size		15	14	15	14
Total No. Dead foetuses		0	0	0	0
Total No. of live foetuses		340	331	360	281
Total weight of live foetuses	Mean	3.74	3.76	3.80	3.62
	SD	0.07	0.07	0.10	0.12
No. of live male foetuses		164	165	172	121
Weight of live male foetuses	Mean	3.83	3.86	3.90	3.70
	SD	0.07	0.07	0.22	0.22
No. of live female foetuses		176	166	188	160
Weight of live female foetuses	Mean	3.64	3.66	3.70	3.50
	SD	0.07	0.21	0.22	0.23
Anogenital Distance (mm)	Mean	2.64	2.70	2.62	2.47
	SD	0.96	0.94	0.90	0.85
Crown Rump Length (cm)	Mean	3.74	3.78	3.78	3.73
	SD	0.27	0.23	0.25	0.32
Sex ratio- Male Female		0.93	0.99	0.91	0.76

 

Summary of Gross Pathological Findings of Dams

No. of Animals/Group: 24

Gross Pathological Findings	Group & Dose (mg/kg)
	G1 (0)	G2 (75)	G3 (150)	G4* (350/300)
No abnormalities detected	22	23	23	20
Lungs: Red discolouration	0	0	0	3
Liver: Mottled	0	0	0	1

Note: * = 1 dam will have more number of observations

Summary of Foetal Observations (Incidence and Percentage)

Observations		Group & Dose (mg/kg b.w.)
		G1 (0)	G2 (75)		G3 (150)	G4 (350/300)
No. of litters examined		22	23		23	20
No. of foetuses examined		340	331		360	281
External Examination
No abnormality Detected	340			330	360	280
Hematoma on right Pinna	0			1(0.3%)	0	0
Lower mandible-Hematoma (Jaw)	0			0	0	1(0.4%)
Visceral Examination
No. of foetus examined for examination		175	171		186	146
No abnormality Detected		173	169		185	142
Pin point Haemorrhages on Lungs		1(0.6%)	0		1(0.5%)	1(0.7%)
Kidney-Pale		1(0.6%)	0		0	0
Kidneys-Haemorrhagic		0	1(0.6%)		0	0
Adrenals-Haemorrhagic		0	1(0.6%)		0	0
Right Kidney- Haemorrhagic		0	0		0	1(0.7%)
Brain – Lateral ventricle Haemorrhagic		0	0		0	2(1.4%)

 

Summary of Foetal Skeletal Observations (Incidence and Percentage)

Skeletal Parameters	G1 (0)		G2 (75)		G3 (150)		G4 (350/300)
Skull:	Foetal. Incidence.	% foetus affected	Foetal. Incidence.	% foetus affected	Foetal. Incidence.	% foetus affected	Foetal. Incidence.	% foetus affected
Parietal incomplete ossification	6	3.6	5	3.1	7	4.0	12	8.9
Interparietal incomplete ossification	1	0.6	1	0.6	0	0	2	1.5
Interparietal Unossified	1	0.6	0	0	0	0	0	0
Supra occipital	0	0	0	0	0	0	1	0.7
Ribs-Thorax:
Wavy ribs in 11^thto14^th	1	0.6	1	0.6	0	0	0	0
Knobby (nodulated) ribs	1	0.6	0	0	0	0	1	0.7
14^th Rib ossification Site(SN)	4	2.4	11	6.9	3	1.7	3	2.2
Thoracic Centrum:
11^th Thoracic centrum dumbbell shaped	1	0.6	0	0	0	0	1	0.7
12^th Thoracic centrum dumbbell shaped	2	1.2	1	0.6	0	0	1	0.7
12^th Thoracic centrum Bipartite ossification	0	0	1	0.6	0	0	0	0
11th Thoracic centrum Bipartite ossification	0	0	0	0	0	0	1	0.7
Vertebral arches:
5th Sternebra dumb bell Shaped	11	6.7	12	7.5	6	3.4	8	5.9
2nd Sternebra UO	10	6.1	7	4.4	6	3.4	4	2.9
2nd Sternebra dumb bell Shaped	3	1.8	4	2.5	3	1.7	3	2.2
Manubrium UO	2	1.2	1	0.6	0	0	0	0
Sternebra UO	1	0.6	2	1.3	0	0	2	1.5
4th Sternebra UO	1	0.6	0	0	1	0.6	2	1.5
5th Sternebra IO	24	14.5	21	13.1	28	16.1	19	14.1
2nd Sternebra IO	8	4.8	5	3.1	11	6.3	6	4.4
4th Sternebra IO	1	0.6	1	0.6	1	0.6	0	0
Sternebra Misaligned	3	1.8	4	2.5	4	2.3	3	2.2
5th Sternebra Bipartite ossification	3	1.8	0	0	3	1.7	1	0.7
6th Sternebra IO	4	2.4	6	3.8	13	7.5	3	2.2
6th Sternebra UO	4	2.4	4	2.5	1	0.6	8	5.9
5th Sternebra UO	16	9.7	14	8.8	11	6.3	11	8.1
Sternebra IO	1	0.6	0	0	1	0.6	4	2.9

 

Skeletal Parameters	G1 (0)		G2 (75)		G3 (150)		G4 (350/300)
Vertebral arches:	Foetal. Incidence.	% foetus affected	Foetal. Incidence.	% foetus affected	Foetal. Incidence.	% foetus affected	Foetal. Incidence.	% foetus affected
5th Sternum IO	1	0.6	0	0	0	0	0	0
6th Sternum IO	1	0.6	0	0	0	0	0	0
6th Sternebra dumb bell Shaped	1	0.6	0	0	1	0.6	0	0
5th Sternebra Misaligned	0	0	1	0.6	0	0	0	0
2nd Sternebra Misaligned	0	0	1	0.6	0	0	0	0
Sternebra dumb bell Shaped	0	0	1	0.6	0	0	0	0
Metatarsal UO	0	0	1	0.6	0	0	0	0
Metatarsal IO	0	0	1	0.6	0	0	0	0
pelvic ventebral arch ossification (extra)	0	0	1	0.6	0	0	0	0
Xiphoid P Sternebra UO	0	0	0	0	3	1.7	1	0.7
Right Ischium UO	0	0	0	0	1	0.6	0	0
4th Sternebra dumb bell Shaped	0	0	0	0	1	0.6	0	0
3rd Sternebra UO	0	0	0	0	1	0.6	0	0
2nd Sternebra B O	0	0	0	0	1	0.6	0	0
2nd Sternebra dumb bell ossification	0	0	0	0	0	0	1	0.7
3rd Sternebra dumb bell ossification	0	0	0	0	0	0	1	0.7
4th Sternebra dumb bell ossification	0	0	0	0	0	0	1	0.7
Pubis IO	0	0	0	0	0	0	1	0.7

 

 

Historical Range

Caesarean Section Data – Gestation Day 20

Parameters	Mean	SD	Min	Max
Females/Study	22.11	5.60	18	25
Pregnant Females	19.33	5.15	16	24
% Pregnant	94.20	6.39	89	98
# All Resorbed	0.00	0.00	0	0
% of Preg. All resorbed	0.00	0.00	0	0
# Corpora Lutea	17.88	4.43	11	18
# Implants	13.88	2.76	9	15
# Resorptions	0.26	0.66	0	4
% Resorptions	3.08	8.56	0	15
# Live Foetuses	13.59	5.91	9	14
# Dead Foetuses	0.01	0.08	0	1
% Dead + Resor./Implants	0.03	0.09	0.0	0.5
Sex Ratio (% Males)	51.43	18.76	0	100
Live Foetal Weights (g)
All foetuses	3.62	0.31	2.79	4.97
Male Foetuses	3.68	0.43	0.00	5.10
Female Foetuses	3.49	0.41	0.00	4.75

Maternal Data – Gestation Day 20

Parameters	Mean	SD	Min	Max
Females/Study	22.18	5.60	18.00	25.00
Pregnant Females	21.03	5.15	16.00	24.00
% Pregnant	94.81	6.39	80.00	96.00
Corrected Maternal Body Weight (g)	235.56	19.24	184.60	295.40
Relative uterus weight (%)	18.31	4.52	2.55	25.07
% Pre implantation loss	5.95	13.53	0.00	26.00
% Post implantation loss	2.01	5.22	0.00	31.00

Foetal External Anomalies

Observations	Foetal Incidence			Litter Incidence
	Incidence		Average %	Incidence	Average %
Anasarca	0.30		0.01	0.20	0.12
Conjoined twins	1.00		0.04	0.03	0.02
Hematoma	1.00		0.04	0.26	0.15
Local edema	0.50		0.02	0.14	0.08
Pallid fetus	0.00		0.00	0.06	0.03
SKULL
Cephalocele	0.03		0.00	0.03	0.02
Craniorachischisis	0.40		0.01	0.20	0.11
Domed head	0.03		0.00	0.03	0.02
Encephalomeninogocele	0.50		0.02	0.03	0.02
Exencephaly	1.00		0.04	0.31	0.18
Meningocele	0.03		0.00	0.03	0.02
Microcephaly	0.03		0.00	0.03	0.02
EYES
Ablepharia	0.22		0.01	0.08	0.05
Anophtalmia	0.52		0.02	0.35	0.21
Cyclopia	0.03		0.00	0.03	0.02
Exophthalmos	0.03		0.00	0.06	0.03
Microphthalmia	2.03		0.07	0.75	0.44
Open eyelid	0.33		0.01	0.17	0.10
EAR
Ear, displaced	0.13		0.00	0.11	0.07
Microtia	0.03		0.00	0.03	0.02
NOSE
Nasal atresia		0.06	0.00	0.06	0.03
Nasal dysplasia		0.10	0.00	0.08	0.05
Rhinocephaly		0.06	0.00	0.06	0.03
MOUTH AND JAW
Aglossia		0.10	0.00	0.08	0.05
Agnathia		0.41	0.01	0.28	0.16
Astomia		0.19	0.01	0.17	0.10
Cleft face		0.06	0.00	0.06	0.03
Cleft lip		0.03	0.00	0.03	0.02
Cleft palate		0.29	0.01	0.25	0.15
High-arched palate		0.13	0.00	0.06	0.03
Microglossia		0.06	0.00	0.06	0.03
Micrognathia		0.31	0.01	0.34	0.20
Microstomia		0.13	0.00	0.11	0.07
Protruding tongue		0.21	0.01	0.31	0.18

Observations	Foetal Incidence		Litter Incidence
	Incidence	Average %	Incidence	Average %
TORSO
Atresia ani	0.29	0.01	0.25	0.15
Gastroschisis	0.22	0.01	0.20	0.11
Holorachischisis	0.03	0.00	0.03	0.02
Omphalocele	0.16	0.01	0.14	0.08
Scoliosis	0.03	0.00	0.03	0.02
Shortened torso	0.06	0.00	0.06	0.03
Spina bifida aperta	0.13	0.00	0.11	0.07
Umbilical hernia	0.34	0.01	0.34	0.20
EXTERMITIES
Clubbed forefoot	0.06	0.00	0.06	0.03
Clubbed hindfoot	0.03	0.00	0.03	0.02
Extrodactyly	0.03	0.00	0.03	0.02
Extromelia	0.03	0.00	0.03	0.02
Forelimb Flexion	0.13	0.00	0.06	0.03
Micromelia	0.10	0.00	0.08	0.05
Phocomelia	0.03	0.00	0.03	0.02
Tail malformation	0.76	0.03	0.99	0.58
ADDITIONAL FINDINGS
Abdominal hernia	0.03	0.00	0.03	0.02
partial twinning	0.03	0.00	0.03	0.02
Shiny	0.10	0.00	0.08	0.05
Polymelia	0.03	0.00	0.03	0.02
Growth retarded	0.86	0.03	0.51	0.30
Fused placenta	0.13	0.00	0.06	0.03
Vibrissal pad, malf.	0.03	0.00	0.03	0.02
Upper jaw missing	0.03	0.00	0.03	0.02
Hard palate missing	0.03	0.00	0.03	0.02

 

Foetal Head Razor Section Anomalies

Observations	Foetal Incidence		Litter Incidence
	Incidence	Average %	Incidence	Average %
Third Ventricle: Slightly Dilated	11	2.50	18	2.48
Third Ventricle: Dilated	6	1.17	8	1.45
Third Ventricle: Misshapen	1	0.39	1	0.56
Lateral Ventricles: Slightly Dilated	18	5.28	49	7.91
Lateral Ventricles: Dilated	7	3.32	20	4.06
Lateral Ventricles: Misshapen	1	0.19	1	0.56
Cerebral Hemisphere: Hemorrhagic	1	0.38	1	0.42
Lateral Ventricles: Hemorrhagic	1	0.25	2	0.37
Nasal Septum: Misshapen	1	0.25	1	0.14
Nasal Conchae: Small	1	0.38	3	0.68
Nasal Cavity: Hemorrhagic	1	0.13	3	0.71
Nasal Cavity: Large	1	0.13	1	0.37
Nasal Cavity: Dilated	1	0.13	1	0.08
Nasal Cavity: Misshapen	1	0.13	1	0.09
Nasal Septum: Hemorrhagic	1	0.13	1	0.12
Eye: Anopthalmia	1	0.13	1	0.37
Eye: Small	1	0.13	1	0.14
Olfactory Lobes: Large	2	0.25	2	0.37
Olfactory Lobes: Misshapen	1	0.13	1	0.19
Palate: Hemorrhagic	1	0.13	1	0.08
Retina (U): Misshapen	1	0.13	2	0.31

Foetal Visceral Anomalies

Observations	Foetal Incidence (%)						Litter Incidence (%)
	Mean		SD		Max		Mean		SD		Max
CORONAL SECTIONS
Cerebrum: Misshapen	0.011		0.06		0.84		0.053		0.57		4.21
Cerebral ventricular enlargement	0.110		0.27		2.58		0.561		2.27		22.02
Hydrocephalus	0.016		0.09		0.73		0.086		0.61		5.73
Lens: Agenesis	0.026		0.18		0.57		0.126		0.54		3.72
Lens: Hypoplastic	0.013		0.07		0.79		0.122		0.84		5.61
Nasal cavity: Enlarged	0.002		0.02		0.48		0.022		0.28		3.93
Retina: Agenesis	0.006		0.06		0.68		0.051		0.43		4.46
Retina: Folded	0.019		0.13		1.47		0.179		0.58		7.03
Microphthalmia	0.019		0.13		1.27		0.207		1.08		9.02
GENERAL
Abdominal cavity: Fluid-filled	0.003		0.04		0.61		0.027		0.38		3.51
Situs inversus	0.009		0.18		0.63		0.118		0.75		3.07
BLOOD VESSLES
Aortic arch: Enlarged	0.002		0.02		0.58		0.032		0.29		2.75
Aortic arch: Interrupted	0		0		0		0		0		0
Aortic arch: Retroesophageal	0.001		0.00		0.26		0.027		0.41		2.89
Azygos vein: Doubled	0.039		0.27		1.72		0.165		0.89		9.27
Azygos vein: Right-sided	0.003		0.02		0.59		0.013		0.24		4.17
Carotid: Abnormal origin	0.004		0.07		0.82		0.027		0.46		2.81
Ductus arteriosus: Hypoplastic	0.004		0.08		0.49		0.037		0.46		3.72
Innominate: Agenesis	0.054		0.40		2.21		0.276		1.27		12.54
Innominate: Hypoplastic	0.009		0.05		1.03		0.046		0.45		5.13
Pulmonary trunk: Stenosis	0.003		0.03		0.74		0.034		0.45		3.81
Truncus arteriosus: Persistent	0.006		0.04		0.81		0.044		0.38		4.74
HEART
Atrium: Enlarged		0.005		0.04		0.57		0.043		0.36		2.98
Cardiomegaly		0.005		0.03		0.52		0.031		0.27		5.33
Dextrocadia		0.003		0.06		0.48		0.026		0.43		4.21
Double apex		0.004		0.04		0.41		0.036		0.25		5.12
Misshapen heart		0.017		0.18		0.73		0.127		0.55		3.91
Situs inversus, cardio-vascular		0.008		0.04		0.56		0.088		0.41		4.49
Ventricular hypertrophy		0.005		0.04		0.68		0.039		0.27		3.00
Ventricular hypoplasia		0.004		0.08		0.53		0.018		0.29		3.12
Ventricular septal defect (VSD)		0.012		0.16		0.71		0.116		0.53		4.93
LUNG AND TRACHEA
Lung: Abnormal lobe formation		0.009		0.08		0.53		0.061		0.52		3.31
Lung: Agenesis		0.006		0.05		0.59		0.048		0.56		4.78
Lung: Diff. hemorrh. (mottled)		0.005		0.04		0.66		0.031		0.46		2.81
Lung: Enlarged		0.003		0.03		0.59		0.021		0.43		3.00
Trachea: Short		0.005		0.05		0.84		0.031		0.27		3.81
DIAPHRAGM
Diaphragmatic hernia		0.009		0.04		0.51		0.063		0.41		5.27

Observations	Foetal Incidence (%)						Litter Incidence (%)
	Mean		SD		Max		Mean	SD	Max
DIGESTIVE TRACT
Esophagus: Short	0.007		0.06		0.87		0.033	0.42	4.81
Stomach: Situs inversus	0.006		0.05		0.48		0.061	0.36	4.00
SPLEEN
Spleen: Hypoplastic	0.004		0.05		0.56		0.031	0.27	3.89
ADRENAL
Focally hemorrhagic	0.009		0.08		0.71		0.065	0.54	4.73
KIDNEY
Distended renal pelvis	0.581		1.22		5.83		5.017	6.22	41.81
Hydronephrosis	0.161		0.47		5.36		0.631	2.18	19.75
Kidney: Diffuse hemorrhagic	0.019		0.17		1.59		0.062	0.84	7.30
Kidney: Displaced	0.006		0.05		0.38		0.059	0.48	4.68
Kidney: Hypoplas. (red. Papilla)	0.822		2.13		12.27		2.167	7.81	43.02
Kidney: Supernumerary	0.003		0.04		0.49		0.021	0.58	3.89
URETER AND BLADDER
Hydroureter	0.347		73.00		3.86		1.223	3.84	17.83
Ureter: Convoluted	0.581		1.99		18.27		3.193	6.93	97.28
Ureter: Distended	1.284		3.71		37.25		6.315	21.84	81.92
Urinary bladder: Enlarged	0.165		0.71		6.73		0.573	1.88	31.36
MALE REPRODUCTIVE TRACT
Testis: Agenesis	0.005		0.10		2.17		0.039	0.44	3.92
Testis: Displaced	0.426		1.89		12.63		2.113	6.83	69.82
Testis: Supernumerary	0.003		0.04		0.81		0.061	0.73	4.93
SKULL
Bone island		0.002		0.02		0.38	0.021	0.28	4.81
Frontals: Contain holes		0.006		0.09		1.16	0.041	0.58	6.13
Frontals: Hypoplastic		0.004		0.05		0.57	0.029	0.37	4.21
Frontals: Misshapen		0.004		0.04		0.56	0.041	0.82	3.68
Frontals: Bifid		0.002		0.03		0.51	0.029	0.36	3.49
Mandible: Agenesis		0.017		0.06		0.62	0.051	0.37	3.28
Mandible: Hypoplastic		0.003		0.02		0.71	0.051	0.28	3.29
Mandible: Misshapen		0.004		0.02		0.52	0.061	0.20	4.04
Maxilla: Agenesis		0.005		0.08		0.83	0.029	0.42	3.29
Maxilla: Misshapen		0.009		0.04		0.52	0.058	0.62	4.74
Nasals: Misshapen		0.006		0.05		0.78	0.059	0.73	5.82
Orbits: Hypoplastic		0.007		0.09		0.65	0.034	0.54	4.71
Orbits: Misshapen		0.033		0.18		0.98	0.339	1.34	7.83
Parietals: Misshapen		0.003		0.02		0.47	0.031	0.31	2.48
SPINAL COLUMN
Vertebral Scrambling		0.018		0.13		1.92	0.078	0.58	9.02
Cervical vertebrae: Fused		0.013		0.17		1.21	0.089	0.53	3.71
Cervical vertebrae: Hypoplastic		0.005		0.03		0.41	0.062	0.47	3.81
Cervical vertebrae: Misaligned		0.004		0.06		0.58	0.027	0.36	4.00
Cervical vertebrae: Misshapen		0.002		0.02		0.34	0.032	0.41	4.86

Observations	Foetal Incidence (%)			Litter Incidence (%)
	Mean	SD	Max	Mean	SD	Max
SPINAL COLUMN
Cervical centra: Split	0.003	0.05	0.79	0.031	0.47	4.89
Thoracic vertebrae: Agenesis	0.061	0.28	3.79	0.223	1.37	9.63
Thoracic vertebrae: Fused	0.024	0.16	1.29	0.126	0.68	4.82
Thoracic vertebrae: Misaligned	0.005	0.06	0.42	0.037	0.51	5.21
Thoracic vertebrae: Misshapen	0.017	0.05	0.61	0.169	0.71	6.82
Thoracic centra: Dumbbell-shaped	0.397	1.07	17.24	1.212	5.81	57.33
Thoracic centra: Split	0.537	1.00	7.82	2.116	4.84	23.69
Thoracic centra: Misshapen	0.010	0.03	0.73	0.067	0.52	3.62
Lumbar vertebrae: Agenesis	0.014	0.13	0.96	0.005	0.03	0.23
Lumbar vertebrae: Fused	0.008	0.07	1.27	0.057	0.59	6.17
Lumbar vertebrae: Misaligned	0.008	0.06	0.80	0.058	0.63	2.27
Lumbar centra: Dumbbell-shaped	0.016	0.07	0.82	0.053	0.58	3.25
Lumbar centra: Split	0.021	0.11	1.89	0.213	0.88	4.82
Sacral vertebrae: Agenesis	0.003	0.04	0.37	0.039	0.03	3.29
Sacral vertebrae: Fused	0.181	0.53	9.67	0.314	2.17	27.73
Sacral vertebrae: Misaligned	0.026	0.18	0.76	0.206	1.45	9.03
Caudal vertebrae: Agenesis	0.007	0.04	0.44	0.063	0.55	3.26
Caudal vertebrae: Fused	0.081	1.45	10.60	0.154	1.02	23.27
Lumbosacral shift	0.037	0.19	1.41	0.312	1.56	9.81
Missing vertebra	0.011	0.04	0.52	0.161	0.49	3.28
Vertebral count > or < normal	0.137	0.25	1.22	0.761	1.39	10.52
STERNEBRAE
Multiple fusions	0.069	0.33	1.23	0.537	1.69	3.82
Agenesis	0.009	0.10	0.27	0.096	0.53	2.18
Split	0.036	0.33	0.82	0.216	1.33	9.54
Misaligned	0.361	1.47	3.81	2.367	2.81	22.17
Asymmetric	0.053	0.18	1.37	0.531	1.67	13.89
Misshapen	0.037	0.11	0.84	0.167	1.84	4.21
RIBS
Agenesis	0.021	0.17	0.29	0.425	1.81	3.72
Cervical	0.516	1.27	4.04	3.482	5.63	17.03
Fused	0.081	0.11	0.37	0.483	1.07	3.29
Hypoplastic	1.005	2.63	16.92	3.760	9.53	53.72
Misshapen	0.884	2.16	5.61	2.190	2.15	37.83
Supernumerary	2.167	3.27	16.83	23.445	19.94	96.81
Wavy	0.213	0.27	1.31	1.067	1.13	9.83
Bent	0.037	0.44	1.21	0.387	2.13	9.52
PELVIC GIRDLE
Ischium: Hypoplastic	0.023	0.04	0.29	0.171	0.59	3.79
SHOULDER GIRDLE
Scapula: Misshapen	0.053	0.038	0.47	0.169	0.733	3.45

 

Observations	Foetal Incidence (%)			Litter Incidence (%)
	Mean	SD	Max	Mean	SD	Max
APPENDICULAR SKELETON
Fore limb
Long bone: Hypoplastic	0.033	0.11	0.83	0.211	1.23	10.01
Long bone: Misshapen	0.002	0.02	0.13	0.063	0.42	3.76
Phalanx: Misaligned	0.007	0.07	0.76	0.059	0.74	3
Phalanx: Clubbed	0.023	0.13	1.27	0.231	0.72	6.03
Hind limb
Long bone: Small	0.034	0.11	1.27	0.236	0.87	5.92
Long bone: Misshapen	0.006	0.03	0.34	0.237	0.59	2.47
Phalanx: Agenesis	0.104	0.61	2.74	0.481	3.22	21.63
Phalanx: Supernumerary	0.019	0.12	1.05	0.116	2.17	4.72
Phalanx: Misaligned	0.011	0.11	0.87	0.053	0.27	3.37
Talus: Advanced	0.025	0.23	1.81	0.095	0.52	2.91

Foetal Skeletal Anomalies

Observations	Foetal Incidence (%)			Litter Incidence (%)
	Mean	SD	Max	Mean	SD	Max
SKULL
Bone island	0.002	0.02	0.38	0.021	0.28	4.81
Frontals: Contain holes	0.006	0.09	1.16	0.041	0.58	6.13
Frontals: Hypoplastic	0.004	0.05	0.57	0.029	0.37	4.21
Frontals: Misshapen	0.004	0.04	0.56	0.041	0.82	3.68
Frontals: Bifid	0.002	0.03	0.51	0.029	0.36	3.49
Mandible: Agenesis	0.017	0.06	0.62	0.051	0.37	3.28
Mandible: Hypoplastic	0.003	0.02	0.71	0.051	0.28	3.29
Mandible: Misshapen	0.004	0.02	0.52	0.061	0.20	4.04
Maxilla: Agenesis	0.005	0.08	0.83	0.029	0.42	3.29
Maxilla: Misshapen	0.009	0.04	0.52	0.058	0.62	4.74
Nasals: Misshapen	0.006	0.05	0.78	0.059	0.73	5.82
Orbits: Hypoplastic	0.007	0.09	0.65	0.034	0.54	4.71
Orbits: Misshapen	0.033	0.18	0.98	0.339	1.34	7.83
Parietals: Misshapen	0.003	0.02	0.47	0.031	0.31	2.48
SPINAL COLUMN
Vertebral Scrambling	0.018	0.13	1.92	0.078	0.58	9.02
Cervical vertebrae: Fused	0.013	0.17	1.21	0.089	0.53	3.71
Cervical vertebrae: Hypoplastic	0.005	0.03	0.41	0.062	0.47	3.81
Cervical vertebrae: Misaligned	0.004	0.06	0.58	0.027	0.36	4.00
Cervical vertebrae: Misshapen	0.002	0.02	0.34	0.032	0.41	4.86
Cervical centra: Split	0.003	0.05	0.79	0.031	0.47	4.89
Thoracic vertebrae: Agenesis	0.061	0.28	3.79	0.223	1.37	9.63
Thoracic vertebrae: Fused	0.024	0.16	1.29	0.126	0.68	4.82
Thoracic vertebrae: Misaligned	0.005	0.06	0.42	0.037	0.51	5.21
Thoracic vertebrae: Misshapen	0.017	0.05	0.61	0.169	0.71	6.82
Thoracic centra: Dumbbell-shaped	0.397	1.07	17.24	1.212	5.81	57.33
Thoracic centra: Split	0.537	1.00	7.82	2.116	4.84	23.69
Thoracic centra: Misshapen	0.010	0.03	0.73	0.067	0.52	3.62
Lumbar vertebrae: Agenesis	0.014	0.13	0.96	0.005	0.03	0.23
Lumbar vertebrae: Fused	0.008	0.07	1.27	0.057	0.59	6.17
Lumbar vertebrae: Misaligned	0.008	0.06	0.80	0.058	0.63	2.27
Lumbar centra: Dumbbell-shaped	0.016	0.07	0.82	0.053	0.58	3.25
Lumbar centra: Split	0.021	0.11	1.89	0.213	0.88	4.82
Sacral vertebrae: Agenesis	0.003	0.04	0.37	0.039	0.03	3.29
Sacral vertebrae: Fused	0.181	0.53	9.67	0.314	2.17	27.73
Sacral vertebrae: Misaligned	0.026	0.18	0.76	0.206	1.45	9.03
Caudal vertebrae: Agenesis	0.007	0.04	0.44	0.063	0.55	3.26
Caudal vertebrae: Fused	0.081	1.45	10.60	0.154	1.02	23.27
Lumbosacral shift	0.037	0.19	1.41	0.312	1.56	9.81
Missing vertebra	0.011	0.04	0.52	0.161	0.49	3.28
Vertebral count > or < normal	0.137	0.25	1.22	0.761	1.39	10.52
STERNEBRAE
Multiple fusions	0.069	0.33	1.23	0.537	1.69	3.82
Agenesis	0.009	0.10	0.27	0.096	0.53	2.18

Observations	Foetal Incidence (%)			Litter Incidence (%)
	Mean	SD	Max	Mean	SD	Max
STERNEBRAE
Split	0.036	0.33	0.82	0.216	1.33	9.54
Misaligned	0.361	1.47	3.81	2.367	2.81	22.17
Asymmetric	0.053	0.18	1.37	0.531	1.67	13.89
Misshapen	0.037	0.11	0.84	0.167	1.84	4.21
RIBS
Agenesis	0.021	0.17	0.29	0.425	1.81	3.72
Cervical	0.516	1.27	4.04	3.482	5.63	17.03
Fused	0.081	0.11	0.37	0.483	1.07	3.29
Hypoplastic	1.005	2.63	16.92	3.760	9.53	53.72
Misshapen	0.884	2.16	5.61	2.190	2.15	37.83
Supernumerary	2.167	3.27	16.83	23.445	19.94	96.81
Wavy	0.213	0.27	1.31	1.067	1.13	9.83
Bent	0.037	0.44	1.21	0.387	2.13	9.52
PELVIC GIRDLE
Ischium: Hypoplastic	0.023	0.04	0.29	0.171	0.59	3.79
SHOULDER GIRDLE
Scapula: Misshapen	0.053	0.038	0.47	0.169	0.733	3.45
APPENDICULAR SKELETON
Forelimb
Long bone: Hypoplastic	0.033	0.11	0.83	0.211	1.23	10.01
Long bone: Misshapen	0.002	0.02	0.13	0.063	0.42	3.76
Phalanx: Misaligned	0.007	0.07	0.76	0.059	0.74	3
Phalanx: Clubbed	0.023	0.13	1.27	0.231	0.72	6.03
Hindlimb
Long bone: Small	0.034	0.11	1.27	0.236	0.87	5.92
Long bone: Misshapen	0.006	0.03	0.34	0.237	0.59	2.47
Phalanx: Agenesis	0.104	0.61	2.74	0.481	3.22	21.63
Phalanx: Supernumerary	0.019	0.12	1.05	0.116	2.17	4.72
Phalanx: Misaligned	0.011	0.11	0.87	0.053	0.27	3.37
Talus: Advanced	0.025	0.23	1.81	0.095	0.52	2.91

Applicant's summary and conclusion

Conclusions:

Pregnant Wistar rats when treated with ‘Genamin 3920’ at 75, 150 and 350/300 mg/kg/day did not reveal any foetal toxicity. No teratogenic effects attributable to treatment were observed. Significant decrease in body weight and feed consumption along with significant decrease in uterine weights were observed in dams treated at 350/300 mg/kg.
Based on findings, the NOAEL of ‘Genamin 3920’ in pregnant Wistar rats is 150 mg/kg for maternal toxicity and 300 mg/kg for foetal toxicity under the tested conditions.

Executive summary:

This study was conducted to estimate the prenatal developmental toxicity and maternal toxicity of Genamin 3920, when administered by oral route to pregnant Wistar rats during Gestation Days (GD) 5 to 19. The study was carried out in accordance with the OECD Guideline for Testing of Chemicals No. 414 and United States EPA Health Effects Test Guideline OCSPP 870.3700.

A total of four groups (G1, G2, G3 and G4) of pregnant Wistar rats were used in the study out of which, Group 1 (G1) served as the vehicle control. Group 2 (G2), Group 3 (G3) and Group 4 (G4) were administered the test item at the dose levels of 75, 150 and 350/300 mg/kg/day respectively, through oral route, once daily, from gestation days (GD) 5 to 19. As mortality was observed at the dose level of 350 mg/kg/day (high dose) during different gestation days, the dose was reduced to 300 mg/kg/day for this group. The control group was administered the vehicle alone (corn oil).

Test item formulations were prepared in corn oil. Dose formulation analysis was performed on formulation samples prepared on the first day of dose administration. The concentration of Genamin 3920 in the dose formulations were in the range between 97.5% - 105.1%, which were within the acceptance criteria (85% - 115%). All the formulations met the acceptance criteria and confirmed that the animals received the intended dose of test item. 

In-life observations included morbidity, mortality, clinical signs of toxicity, maternal body weight and food consumption. A Caesarean section was performed on Day 20 of gestation followed by which, visceral examination of all the dams was performed. Litter data, sex and weight of individual foetuses were recorded followed by which, external malformations, visceral, head razor and skeletal examinations were performed on foetuses.

All dams belonging to control, low and mid dose were normal and were free from all visible clinical signs throughout the treatment period. Three dams (Dam No. T018/074, T018/084 and T018/086) treated at 350 mg/kg body weight/day were found dead between GD 9-15. Animals treated at 350 mg/kg/day displayed salivation, burrowing behaviour, hyperactivity and aggression. The observed clinical signs disappeared when the high dose was reduced to 300 mg/kg/day.

Maternal body weight, body weight gain and feed consumption data were significantly reduced from GD 8-20 at the dose of 350/300 mg/kg. All dams belonging to the control, low and mid dose treated groups displayed normal body weight gain throughout the study period. Adjusted maternal weight (after exclusion of uterine weight) of treated dams derived on GD 20 was significantly reduced in the 350/300 mg/kg group. No significant changes were observed in the low and mid dose and was comparable to control group animals.

There were no changes in T3, T4 and TSH values attributable to treatment. No pathological observations in thyroid/parathyroid were evident in the control, low, mid and high dose treated dams.

In the found dead dams belonging to high dose, gross pathological evaluations revealed red discoloration (3/3) in lung and mottled liver (1/3) which were considered to be treatment related. All other dams belonging to high dose group following reduction of dose from 350 to 300 mg/kg/day survived the entire treatment period and displayed no gross pathological abnormalities. No abnormalities were detected in the mid and low dose treated dams as well as the control animals during gross pathological examinations.

Histopathological observations of the found dead animals revealed kidneys with necrosis, bilateral, multifocal (3+) in 2 dams and kidneys with necrosis, bilateral, multifocal (2+), hyperplasia with squamous cell, non-glandular stomach, diffuse (3+) in one dam in the high dose treated group of 350 mg/kg body weight/day. No histopathological lesions in thyroid/parathyroid were recorded in the survived animals of high dose as well as control, low and mid dose.

No abnormalities were observed in the evaluated maternal parameters viz., mean gravid absolute and relative uterine weights, placental weight, number of corpora lutea, No of implantations, resorptions and implantation losses in treated (75 and 150mg/kg/day) and control group. Significant changes were observed in the gravid uterine weights of high dose treated group (350/300 mg/kg/day) as compared to the control group.

There were no significant changes in the mean number of male and female foetuses and total number of foetuses in any of the treated groups (75, 150 and 300 mg/kg/day) as compared to the control group. Foetal weight was comparable between treated and control groups. No malformations related to treatment were detected during external examination of the pups.

No malformations/abnormalities were detected in any of the treated and control groups during the visceral and head razor examinations.

Foetal skeletal examination did not reveal any treatment related malformations/ abnormalities in any of the treated and control groups.

Conclusion

Pregnant Wistar rats when treated with ‘Genamin 3920’ at 75, 150 and 350/300 mg/kg/day did not reveal any foetal toxicity. No teratogenic effects attributable to treatment were observed. Significant decrease in body weight and feed consumption along with significant decrease in uterine weights were observed in dams treated at 350/300 mg/kg.

Based on findings, the NOAEL of ‘Genamin 3920’ in pregnant Wistar rats is 150 mg/kg for maternal toxicity and 300 mg/kg for foetal toxicity under the tested conditions.