Ammonium 2-amino-4-(hydroxymethylphosphinyl)butyrate

Repeated dose toxicity: oral

Administrative data

Endpoint:

chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

May 1983 - Jun 1984

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1984

Report date:

1984

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 452 (Chronic Toxicity Studies)

Version / remarks:

May 12, 1981

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Ammonium 2-amino-4-(hydroxymethylphosphinyl)butyrate

EC Number:

278-636-5

EC Name:

Ammonium 2-amino-4-(hydroxymethylphosphinyl)butyrate

Cas Number:

77182-82-2

Molecular formula:

C5H12NO4P.H3N

IUPAC Name:

ammonium 2-amino-4-[hydroxy(methyl)phosphoryl]butanoate

Test animals

Species:

dog

Strain:

Beagle

Details on species / strain selection:

This system has been selected as an internationally recognized standard non-rodent species.

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: KFM, Kleintierfarm Madoerin AG, 4414 Fuellinsdorf / Switzerland
- Age at study initiation: 4 - 6 months
- Weight at study initiation: 4.2 - 8.0 kg (males), 3.6 - 6.5 kg (females)
- Housing: individually
- Diet (e.g. ad libitum): each dog was provided with 300 grams of repelleted standard Kliba no. 335 dog maintenance diet for 3 hours daily
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: 17 days under test conditions, after veterinary examination

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/- 2 °C
- Humidity (%): 55 +/- 10 %
- Air changes (per hr): 20
- Photoperiod (hrs dark / hrs light): 12 h / 12 h

Administration / exposure

Route of administration:

oral: feed

Details on route of administration:

Peroral ingestion (feeding) was considered the most appropriate route for testing systemic effects of the test article. Therefore, each dog received daily 300 grams of the appropriate test article / diet mixture for the duration of the study. The animals of group 1 (controls) received similar diet without the test article. The feed was available daily for approximately 3 hours. Any remaining feed was weighed to calculate the daily food consumption.

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

DIET PREPARATION
- Rate of preparation of diet (frequency): twice monthly
- Mixing appropriate amounts with (Type of food): standard Kliba no. 335 dog maintenance diet
- Storage temperature of food: room temperature

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The stability of the test substance was tested before starting the study. The homogeneity and concentration of the test article in the diet preparations were determined every three months in the analytical laboratory of RCC.

Duration of treatment / exposure:

12 months

Frequency of treatment:

daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

2 mg/kg bw/day (nominal)

Doses / concentrations 2

Dose / conc.:

5 mg/kg bw/day (nominal)

Doses / concentrations 3

Dose / conc.:

8.5 mg/kg bw/day (nominal)

No. of animals per sex per dose:

8

Control animals:

yes, plain diet

Details on study design:

- Dose selection rationale: The dietary route administration was selected in accordance with regulatory requirements for the registration of pesticides to define human health risk.

Examinations

Observations and examinations performed and frequency:

MORTALITY / VIABILITY: Yes
- Time schedule: at least twice daily

CLINICAL OBSERVATIONS: Yes
- Time schedule: at least twice daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Time schedule: daily

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: at pretest and after 3, 6, 9 and 12 months
- Dose groups that were examined: all animals

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at pretest and after 1, 3, 6 and 12 months of treatment
- Anaesthetic used for blood collection: No
- Animals fasted: Not specified
- How many animals: all animals
- Parameters examined: erythrocyte count, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, platelet count, reticulocyte count, nucleated erythrocytes - normoblasts, total leukocyte count, differential leukocyte count, red cell morphology, thromboplastin time, partial thromboplastin time, thrombin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at pretest and after 1, 3, 6 and 12 months of treatment
- Animals fasted: Not specified
- How many animals: all animals
- Parameters examined: glucose, urea, creatinine, uric acid, total bilirubin, direct bilirubin, total cholesterol, aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, creatine kinase, alkaline phosphatase, gamma-glutamyl-transferase, ornithine carbamyl-transferase, calcium, phosphorus, sodium, potassium, chloride, total protein, protein electrophoresis, bromosulfophthalein, phenolsulfonphthalein

URINALYSIS: Yes
- Time schedule for collection of urine: at pretest and after 1, 3, 6 and 12 months of treatment
- Metabolism cages used for collection of urine: No
- Animals fasted: Not specified
- Parameters examined: specific gravity, color, appearance, pH, protein, glucose, ketone, bilirubin, blood, nitrite, urobilinogen, urine sediment

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No

OTHER:
- Hearing Tests: at pretest and after 3, 6, 9 and 12 months
- Examinations of the teeth and mucous membranes: at pretest and after 3, 6, 9 and 12 months
- Electrocardiograms: at pretest, after 6 months and after 12 months
- Bone marrow examinations

Sacrifice and pathology:

GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes

Statistics:

The following statistical methods were used to analyze body weights, food consumption, organ weights and clinical laboratory data:
- Univariate one-way analysis of variance was used to assess the significance of intergroup differences if the variables could be assumed to follow a normal distribution. Student's t-test, based on a pooled variance estimate, was used for intergroup comparisons (i.e. single treatment groups against the control group).
- William's test was used to determine the lowest dose group significantly different from the control group, if a monotone (increasing or decreasing) dose-response relationship existed.
- The difference of the overall mean of the treated groups to the control group was tested for significance if some evidence existed for differences between the treatment groups and the control group, and a monotone dose-response relationship and significant t-test were absent. This is referred to as "significance as a whole" in the results section.
- The median test was used to assess the significance of intergroup differences if the variables were defined on a discrete scale.
- Fisher's exact test for 2 x 2 tables was applied if the variables could be dichotomized without loss of information.
The adjustment for multiple testing was not performed in a formal way (i.e. Bonferoni inequality and subsequently lowering the test level), but rather informally by comparing the expected and observed number of significant results obtained.
Individual values, means, standard deviations and t-statistics were rounded off before printing. For example, t-tests were calculated on the basis of exact values for means and pooled variances and then rounded off to two decimal places. Therefore, two groups may display the same printed means for a given parameter, yet display different t-statistic values with respect to the control group.

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Clinical symptoms were noted in three animals, manifested by trismus, salivation, hyperactivity followed by somnolence and hypoactivity, as well as stereotypic stiff gait, tremor, ataxia, whining, urinating, tonic-clonic spasms, paddling movements, opisthotonus and lateral recombency. These symptoms were observed in male No. 28 on days 9 - 12 and 14, and in females No. 61 on days 9 - 10 and in No. 63 on day 9. Generally, symptoms were noted from 0 - 2 hours after administration. However, in male No. 28, these symptoms were seen 23 hours after the administration on days 11 and 12. Convulsive movements, when observed, persisted for 1 to 5 minutes. Three hours prior to death, male No. 28 showed increased heart rate (> 200 beats/min), dyspnea, decreased body temperature (36.4 degrees centrigrade), reduced corneal reflexes and severe apathy.
In dog No. 47 of group 2, slightly hyperemic areas were noted on the skin of the axillary and abdominal regions, and on the skin of the stifle during weeks 15 - 21. Several times during that seven weeks, the hyperemia observed in the axillary region was more pronounced.
Dog No. 49 of group 3 revealed local alopecia on the ear lobes and alopecia as well as hyperemic skin on all paws from sixth months onwards. After 12 months, alopecia on the paws were widened, and additionally, new areas of slight alopecia were discerned: eye periphery, mandible and thoracic region, the latter also with hyperemic skin.
Both regular and sporadic incidents of soft feces, slight to marked in degree, were observed in all groups. In the males, a reversed dose-related severity was noted (greatest severity in group 1). In the females, the severity was similar in all groups. The feces of some animals from all treated and control groups were observed to contain occasionally traces of mucous and / or blood, but no differences between any group were noted. Sporadic vomiting was noted in some dogs of each group.

Mortality:

mortality observed, treatment-related

Description (incidence):

In group 4, male No. 28 died on day 14, and female No. 63 was found dead on the morning of day 10.
For the two and one days preceding the spontaneous deaths of these animals, respectively, they were observed to have consumed rapidly the entire 300 gram daily feed ration, resulting in abruptly increased test article consumption, whereas they consumed considerably less food during the first 11 or 8 days of treatment, respectively. It is this rapid food (and therefore test article) consumption that is considered to be the cause of the symptoms described below. In dog No. 61 which survived, similar symptoms were observed prior to offering diet but the animal showed almost no food consumption on the day which symptoms were noted and highly decreased food consumption in the days following, although it increased slightly each day. After the initial observation of symptoms on day 9, no further symptoms were noted.
These findings indicate that these animals show individual sensitivity to the high doses of the test article, which seems to demonstrate a narrow dose-response relationship.
All other dogs survived the duration of the study.

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

The mean body weight gains of the treated males, especially group 4, was slightly reduced during the first six months of treatment, when compared with those of the control group.
The abrupt decline of body weight seen in four group 3 male dogs at week 11 was inexplicable, but because of its transient nature, it was considered not to be related to treatment. After 27 weeks of study all male groups and the females of groups 1 and 2 showed an abrupt decline. This was due to the slightly lower body weights of the remaining animals after interim sacrifice. In the females, the mean body weight gains of group 4 was slightly increased when compared with those of the control.

Food consumption and compound intake (if feeding study):

effects observed, non-treatment-related

Description (incidence and severity):

With the exception of the group 4 males during week 1, the mean food consumption of the treated males was comparable to that of the controls. In the females, the mean food consumption of group 2 was decreased, whereas groups 3 and 4 were increased when compared with the control group.

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

effects observed, non-treatment-related

Description (incidence and severity):

Somer superficial ocular abnormalities such as small opaque spots, reddened conjunctivae and watery and / or purulent discharge were noted in animals of all groups. These findings were considered to be spontaneous in nature as they are commonly observed in animals of this breed and age.

Haematological findings:

no effects observed

Description (incidence and severity):

The assessment of hematological data indicated no changes of toxicological significance after 1, 3, 6 and 12 months of treatment.
All differences in the results were considered incidental and of normal biological variation.

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

The assessment of clinical biochemistry data indicated no changes of toxicological significance after 1, 3, 6 and 12 months of treatment.
All differences in the results were considered incidental and of normal biological variation.

Urinalysis findings:

no effects observed

Description (incidence and severity):

The assessment of urinalysis data indicated no changes of toxicological significance after 1, 3, 6 and 12 months of treatment.
All differences in the results were considered incidental and of normal biological variation.

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

With the exception of a few statistically significant changes in organ weights and organ weight ratios which were considered to be incidental and of no toxicological relevance, there were no changes in absolute and relative organ weights.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

In the male that died spontaneously, the main gross lesions were increased fluid in the pericardium and subendocardial hemorrhages in the right heart.
In the female that died spontaneously, the main gross findings were numerous gray-green foci in the lung; the lung was not collapsed.
In the dogs killed on schedule, a number of gross lesions were observed at a random incidence in various organs.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Heart
In both dogs that died during the study, multifocal myocardial necrosis was noted. This necrosis was marked and multi-cellular in the male dog (No. 28), with beginning granulocytic infiltration, and it was accompanied by hemorrhages and diffuse microvesicular lipid deposition. In the female (No. 63), the necrosis was slight and mainly unicellular without granulocytic reaction.

Lungs
Marked aspiration of diet material with beginning inflammatory reaction was noted in the female dog (No. 63) that died during the study. Bronchi and bronchioli were markedly filled with foreign material, and the bronchial epithelium was completely desquamated and necrotic.

Stomach
In the female (No. 63) that died during the study, a small acute ulcer was seen in the pyloric mucosa. This lesion corresponded to the red focus described at necropsy.

Other organs / tissues
After the 6-month and the 12-month administration period, a number of common findings were noted in various organs. The type, incidence, and severity of these lesions were similar in the treated dogs and in the controls.

Other effects:

no effects observed

Description (incidence and severity):

- Hearing tests: No change of the auditory perception was noted.
- Examinations of teeth and mucous membranes: No change in the teeth or mucous membranes was noted.
- Electrocardiogram: All findings observed in this study were spontaneous in nature, except for P pulmonale in male No. 26 (group 4) which was possibly test article-related. The cause of the P pulmonale after 12 months treatment in animal No. 26 can not be determined, whether it is test article-related or spontaneous. There were, however, no similar findings in any other animal of this group, so that the P pulmonale could be considered spontaneous (developing tricuspid dysplasia) rather than test article-related. The heart rhythm changes, such as atrioventricular blocks (first and second degree) caused by excessive vagal activity during the respiratory arrhythmia, as well as sinus tachycardia - the most common primary heart arrhythmia - caused by excitation, are findings which occur frequently in clinically healthy Beagle dogs. Two dogs of group 4 which died suddenly on day 10 and 14, were not electrocardiographically examined prior to death. A slight decrease in heart rate within the normal range was observed in groups 2 and 3, more distinct in group 4 after six months of treatment. After 12 months of treatment, no dose related changes in the heart rate could be seen. Statistically significant differences between pretest and 6 or 12 months values in some ECG parameters were of limited biological relevance, because they occurred in control and treated groups. Based on these findings there was no indication of cardiotoxic effects.
- Bone Marrow Examination: The cells of the erythro-, granulo- and thrombopoietic system and the other cells of the animals sacrificed at 6 and 12 months showed no relevant test article-related quantitative or qualitative changes. No biologically relevant differences were noted between the control group and group 4.

Effect levels

1

Dose descriptor:

NOEL

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: absence of effects

2

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: neurological signs, circulatory failure in 2 animals

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

Table 1: Food Consumption (g/animal/day) Treatment / Males

	Group 1	Group 2	Group 3	Group 4
Week 1	266	253	249	218
Week 2	284	271	275	245
Week 3	290	276	287	285
Week 4	291	284	289	286
Week 5	278	279	289	279
Week 6	294	292	291	280
Week 7	295	299	300	289
Week 8	297	298	298	285
Week 9	300	300	300	289
Week 10	292	300	298	283
Week 11	288	300	299	287
Week 12	292	299	300	289
Week 13	298	300	300	285
Week 14	296	300	299	288
Week 15	298	298	300	290
Week 16	297	300	300	289
Week 17	297	299	300	290
Week 18	294	300	300	286
Week 19	295	300	300	291
Week 20	300	300	300	290
Week 21	296	299	300	288
Week 22	298	300	300	289
Week 23	297	300	300	292
Week 24	296	300	300	290
Week 25	292	297	300	284
Week 26	293	300	30	287
Week 27	286	297	300	289
Week 28	294	300	300	281
Week 29	293	300	300	282
Week 30	292	300	300	285
Week 31	293	300	300	276
Week 32	281	300	300	283
Week 33	289	300	300	282
Week 34	289	300	300	285
Week 35	283	300	300	274
Week 36	296	300	300	284
Week 37	286	300	300	287
Week 38	273	300	300	285
Week 39	283	300	300	279
Week 40	293	300	300	278
Week 41	284	300	300	282
Week 42	280	300	300	283
Week 43	290	300	300	281
Week 44	295	300	300	292
Week 45	293	300	300	279
Week 46	285	300	300	280
Week 47	284	300	300	275
Week 48	236	300	289	280
Week 49	268	300	300	281
Week 50	283	300	297	283
Week 51	271	300	300	278
Week 52	275	297	300	278
Week 53	289	300	300	277
Treatment mean	288	297	297	282

Table 2: Food Consumption (g/animal/day) Treatment / Females

	Group 1	Group 2	Group 3	Group 4
Week 1	238	208	243	212
Week 2	256	233	254	244
Week 3	251	236	269	273
Week 4	251	230	285	276
Week 5	253	211	281	268
Week 6	253	240	289	285
Week 7	278	247	288	291
Week 8	269	255	281	282
Week 9	272	268	283	290
Week 10	269	253	284	289
Week 11	281	256	289	286
Week 12	273	251	291	294
Week 13	271	249	288	287
Week 14	281	250	285	289
Week 15	283	260	285	287
Week 16	279	263	289	290
Week 17	280	257	288	291
Week 18	277	257	293	292
Week 19	284	264	292	294
Week 20	287	249	294	291
Week 21	270	241	288	287
Week 22	280	267	292	292
Week 23	284	262	291	298
Week 24	285	256	292	292
Week 25	279	241	283	287
Week 26	272	240	284	293
Week 27	255	233	287	284
Week 28	267	227	291	293
Week 29	260	226	299	294
Week 30	269	217	300	300
Week 31	262	207	300	300
Week 32	266	199	295	283
Week 33	262	205	280	272
Week 34	245	216	282	272
Week 35	260	206	287	288
Week 36	275	225	294	293
Week 37	258	223	292	296
Week 38	264	243	292	298
Week 39	249	221	264	284
Week 40	247	244	288	300
Week 41	231	229	277	295
Week 42	251	230	276	300
Week 43	252	233	283	300
Week 44	259	245	299	300
Week 45	255	221	290	300
Week 46	257	226	289	300
Week 47	256	216	277	300
Week 48	252	218	280	300
Week 49	246	214	281	300
Week 50	245	222	288	300
Week 51	239	208	285	300
Week 52	248	199	271	300
Week 53	244	195	276	300
Treatment mean	263	234	285	289

Table 3: Body Weights (gram) Treatment / Males

	Group 1	Group 2	Group 3	Group 4
Day 3 Week 1	6842	6621	6532	6397
Day 10 Week 2	7118	6844	6641	6361
Day 17 Week 3	7418	7108	6961	6730
Day 24 Week 4	7705	7382	7222	6977
Day 31 Week 5	7814	7533	7495	7095
Day 38 Week 6	8014	7694	7637	7232
Day 45 Week 7	8210	7913	7769	7431
Day 52 Week 8	8294	8084	7953	7461
Day 59 Week 9	8479	8222	8149	7590
Day 66 Week 10	8415	8584	8317	7805
Day 73 Week 11	8712	8500	7780	7896
Day 80 Week 12	8923	8572	8559	8011
Day 87 Week 13	9118	8713	8807	8049
Day 94 Week 14	9209	8763	8789	8149
Day 101 Week 15	9352	8844	8856	8245
Day 108 Week 16	9452	8921	9026	8349
Day 115 Week 17	9551	9094	9082	8443
Day 122 Week 18	9659	9174	9203	8526
Day 129 Week 19	9729	9268	9263	8586
Day 136 Week 20	9842	9364	9343	8611
Day 143 Week 21	9987	9388	9381	8690
Day 150 Week 22	10026	9467	9401	8672
Day 157 Week 23	10055	9502	9505	8763
Day 164 Week 24	10133	9587	9596	8829
Day 171 Week 25	10102	9602	9682	8890
Day 178 Week 26	10229	9718	9690	8910
Day 185 Week 27	10159	9691	9661	8861
Day 192 Week 28	9671	9425	9309	8363
Day 199 Week 29	9767	9383	9306	8457
Day 206 Week 30	9795	9348	9322	8516
Day 213 Week 31	9845	9433	9411	8561
Day 220 Week 32	9876	9448	9485	8703
Day 227 Week 33	9928	9475	9453	8727
Day 234 Week 34	9961	9435	9368	8691
Day 241 Week 35	9929	9552	9377	8725
Day 248 Week 36	9982	9567	9419	8736
Day 255 Week 37	10086	9599	9444	8620
Day 262 Week 38	10036	9625	9516	8879
Day 269 Week 39	10091	9690	9575	8987
Day 276 Week 40	10287	9863	9801	9055
Day 283 Week 41	10281	9785	9839	9158
Day 290 Week 42	10239	9866	9997	9226
Day 297 Week 43	10332	9899	9917	9239
Day 304 Week 44	10244	9898	10004	9304
Day 311 Week 45	10310	9975	10002	9286
Day 318 Week 46	10376	9902	9960	9248
Day 325 Week 47	10497	10008	10204	9270
Day 332 Week 48	10361	10054	10100	9323
Day 339 Week 49	10262	10063	9927	9338
Day 346 Week 50	10130	10058	10045	9347
Day 353 Week 51	10202	10000	10074	9349
Day 360 Week 52	10124	10035	9989	9423
Day 367 Week 53	10138	9970	9929	9434

Table 4: Body Weights (gram) Treatment / Females

	Group 1	Group 2	Group 3	Group 4
Day 3 Week 1	5214	5046	5345	5230
Day 10 Week 2	5520	5210	5538	5538
Day 17 Week 3	5734	5525	5779	5954
Day 24 Week 4	5893	5698	6066	6314
Day 31 Week 5	6060	5656	6268	6540
Day 38 Week 6	6119	5784	6445	6694
Day 45 Week 7	6416	5908	6726	7055
Day 52 Week 8	6553	6163	6788	7054
Day 59 Week 9	6608	6371	6943	7284
Day 66 Week 10	6718	6495	7035	7478
Day 73 Week 11	6902	6616	7141	7589
Day 80 Week 12	6984	6677	7253	7759
Day 87 Week 13	7077	6773	7423	7882
Day 94 Week 14	7203	6801	7446	7980
Day 101 Week 15	7171	6882	7508	8034
Day 108 Week 16	7375	6994	7609	8194
Day 115 Week 17	7458	7110	7650	8344
Day 122 Week 18	7468	7173	7780	8405
Day 129 Week 19	7602	7238	7802	8441
Day 136 Week 20	7645	7261	7853	8489
Day 143 Week 21	7735	7257	7906	8524
Day 150 Week 22	7785	7339	7907	8577
Day 157 Week 23	7863	7384	7976	8651
Day 164 Week 24	7877	7490	8076	8732
Day 171 Week 25	7967	7459	8105	8781
Day 178 Week 26	7980	7475	8157	8882
Day 185 Week 27	7900	7343	8210	8741
Day 192 Week 28	7312	6708	8277	9029
Day 199 Week 29	7388	6867	8332	9033
Day 206 Week 30	7430	6954	8286	9110
Day 213 Week 31	7490	6826	8395	9243
Day 220 Week 32	7559	6735	8413	9225
Day 227 Week 33	7723	6919	8422	9191
Day 234 Week 34	7566	6956	8434	9044
Day 241 Week 35	7580	6912	8435	9062
Day 248 Week 36	7712	7006	8467	9126
Day 255 Week 37	7785	7141	8540	9153
Day 262 Week 38	7782	7225	8532	9183
Day 269 Week 39	7862	7255	8489	9138
Day 276 Week 40	7976	7304	8613	9281
Day 283 Week 41	7844	7241	8531	9304
Day 290 Week 42	7909	7225	8508	9472
Day 297 Week 43	7912	7356	8460	9470
Day 304 Week 44	7906	7344	8562	9463
Day 311 Week 45	7869	7296	8509	9444
Day 318 Week 46	7880	7405	8541	9494
Day 325 Week 47	7977	7324	8544	9513
Day 332 Week 48	7989	7415	8631	9542
Day 339 Week 49	7926	7382	8498	9564
Day 346 Week 50	7927	7538	8502	9589
Day 353 Week 51	7762	7316	8465	9611
Day 360 Week 52	7890	7392	8499	9646
Day 367 Week 53	7845	7305	8507	9743

Table 5: Organ Weights (gram) after 6 months / Males

	Group 1	Group 2	Group 3	Group 4
Body weight	10650	9908	9918	9433
Brain	71.9	77.6	71.4	72.9
Heart	91.1	84.5	82.2	81.7
Liver	283.5	254.8	239.4	238.4
Lung	94.9	81.3	79.6	82.8
Kidneys	46.79	49.18	46.43	51.59
Spleen	43.51	30.82	36.79	28.35
Testes	15.61	14.90	14.23	13.71
Adrenals	1.10	1.07	1.09	1.17
Pituitary	0.064	0.058	0.051	0.065
Thyroid	0.82	0.78	0.70	0.67
Thymus	7.61	5.22	4.04	5.66

Table 6: Organ Weights (gram) after 6 months / Females

	Group 1	Group 2	Group 3	Group 4
Body weight	8015	7348	7810	7850
Brain	66.8	63.9	66.4	69.6
Heart	65.5	66.2	66.7	71.8
Liver	258.9	222.8	252.9	233.8
Lung	63.3	61.8	69.8	73.2
Kidneys	33.93	31.97	34.73	34.43
Spleen	28.72	20.41	22.94	28.01
Ovaries	1.361	1.185	0.962	1.095
Adrenals	1.09	1.12	1.02	0.97
Pituitary	0.050	0.050	0.055	0.047
Thyroid	0.67	0.50	0.65	0.55
Thymus	5.98	5.04	6.19	4.39

Table 7: Organ Weights (gram) after 12 months / Males

	Group 1	Group 2	Group 3	Group 4
Body weight	9688	9300	9275	8913
Brain	76.4	75.4	76.6	80.3
Heart	94.7	90.1	87.7	87.1
Liver	243.9	249.0	244.2	246.5
Lung	76.6	87.3	79.4	71.1
Kidneys	48.34	55.17	45.96	42.88
Spleen	28.72	29.54	24.84	28.31
Testes	13.05	14.86	14.27	14.25
Adrenals	0.95	1.31	1.39	1.26
Pituitary	0.060	0.067	0.057	0.053
Thyroid	0.77	0.90	0.74	0.75
Thymus	7.05	5.80	5.57	4.97

Table 8: Organ Weights (gram) after 12 months / Females

	Group 1	Group 2	Group 3	Group 4
Body weight	7310	6900	8000	9040
Brain	66.2	67.1	72.4	74.1
Heart	66.6	61.6	74.2	78.4
Liver	219.9	199.3	227.3	290.6
Lung	58.6	54.1	72.7	72.7
Kidneys	35.31	31.48	36.40	43.03
Spleen	35.34	26.59	26.99	30.24
Ovaries	0.951	1.299	1.324	1.644
Adrenals	1.19	1.15	1.47	1.22
Pituitary	0.063	0.043	0.063	0.072
Thyroid	0.63	0.62	0.61	0.80
Thymus	4.88	4.48	4.98	7.00