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Some serological characteristics of an anti bovine luteinizing hormone (NIH-LH-B₃) rabbit serum were reported in the previous paper and it was concluded that the serologically specific antiserum to LH was obtained after absorption with bovine serum and bovine thyroid stimulating hormone.
In the present paper some biological properties of an anti bovine LH serum were reported.
The antiserum was obtained by immunizing rabbits with bovine LH by the modified method of Moudgal and Li¹⁶⁾.
The effect of the antiserum on the biological activities of bovine and ovine LH were studied by the method of rat ovarian ascorbic acid depletion described by Parlow¹¹⁾. A group of rats was injected LH and the antiserum while the other, control group, was injected LH and normal rabbit serum. Ovarian ascorbic acid depletion of the rats injected LH and the antiserum was less than that of control. Therefore, it became obvious that biological activities of bovine and ovine LH were inactivated by the antiserum to bovine LH.
The effects of the antiserum on the biological activities of bovine, ovine and porcine gonadotrophins (GTH) were determined by the rabbit ovulation test. Ovulation occured in the control rabbits injected intravenously mixture of rabbit serum and GTH of the each species, but did not appear in the rabbits injected mixture of the antiserum and GTH of the each species. The rasults show that the biological activities of bovine, ovine and porcine GTH were inhibited by the antiserum.
The effects of the antiserum on the biological activities of PMSG and HCG were determined by the rat ovarian weight method. The ovarian weight of rats injected PMSG and the antiserum was less than that of control group. This shows that the biological activities of PMSG was inhibited by the antiserum. As to HCG, however, the ovarian weight of rats injected HCG and the antiserum was not reduced than that of control group. This shows that the biological activities of HCG was not inhibited by the antiserum.
The effects of the antiserum on the biological activities of endogenous GTH in rabbt and rat were examined. Some adult female rabbits were injected the antiserum intravenously and immediately after injection they were mated with male rabbits. Control rabbits were injected a normal rabbit serum instead of the antiserum. None of the rabbits injected the antiserum had ovulation but all of the control rabbits had ovulation. Some intact male rats, 22 days old, were injected the antiserum intrave-nously for 5 days. On the 27th day of birth, they were autopsied and the weights of testis and ventral prostate were measured. The weights of testis and ventral prostate of rats treated with the antiserum were less than that of control rats which received the injection of normal rabbit serum. These results show that the antiserum inhibited the biological activities of endogenous GTH in the rabbit and the rat.

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In our earlier report, it has been shown that the estrous cycle length of the cow can be shortened by inserting rubber tube into the uterine horn during the first half of the cycle. Our recent experiments have demonstrated that the intrauterine injection of a viscous gel-like substance at early luteal phase can accelerate the regression of the bovine corpus luteum and, on the contrary, the treatment at late luteal phase can delay it. In the latter experiments, we have also found that the endometritis is induced temporarily by this treatment. From these results, it is supposed that the inflammatory changes of the endometrium may be related to the life span of the corpus luteum.
The purpose of the work presented here is to investigate 1) the effects of intrauterine infusion of iodine solution as an irritant on the estrous cycle length of the cow and 2) the irritating effects of the treatment on the bovine endometrium.
1) Experiment 1. The effects of intrauterine infusion of iodine solution on the estrous cycle length. Two preparations of iodine i.e. Lugol's solution and solution of polyvinyl pyrrolidone (PVP-I) were used. They were injected into the uterus through the cervix by using a sterilized metal catheter. The treatments were performed on a total of 43 cycles of 11 cows including 9 Japanese beef cows and 2 Jersey cows. When an animal was used successively, she was retreated after it was confirmed that the previous treated cycle had been followed by 1-3 cycles of a normal length. After the treatment, involution of the corpus luteum, development of the follicle and ovulation were examined via rectal palpation at 1-3 days interval. The cervical mucus stained with Giemsa was also examined microscopically. The day on which ovulation occurred was considered as Day 0 of the cycle.
i) Intrauterine infusion of Lugol's solution on Day 3 : Five to fourty ml of Lugol's solution at various concentrations was injected into the uterus of a total of 18 cows on Day 3. The results were shown in Table 1. In 14 of the treated animals, ovulation occurred 6-13 days (av. 8.6±1.9 days) after the treatment and the mean length of the treated cycles was reduced to 11.6±1.9 days. Estrouscycles of the remaining animals were of a normal length in 3 and fairly lengthened in 1. It was also noticed that Lugol's solution diluted 10-20 fold by distilled water was effective enough to modify the cycle length.
ii) Intrauterine infusion of Lugol's solution on Day 16 : In all of 3 animals treated with 40ml of original Lugol's solution on Day 16 (late luteal phase), ovulation occurred 9-11 days (ay. 10.0±1.0 days) after the treatment and the mean length of the treated cycles was lengthened to 26.0±1.0 days.
iii) Intrauterine infusion of PVP-I solution on Day 3 : A total of 21 cows was treated with 1.5-40.0ml of 2.5-100.0mg/ml solution of PVP-I on Day 3. The results were shown in Table 2. In 15 animals injected with 2.5-40.0ml of PVP-I solution, ovulation occurred 6-10 days (av, 7.8±1.3 days) after the treatment and the mean length of the treated cycles was reduced to 10.8±1.3 days. Estrous cycles of the remaining 6 animals were of a normal length. PVP-I solution at the concentration of at least 10mg/ml was noticed to be effective to modify the cycle length.
iv) Responses of cervical mucus to the intrauterine treatment : On 19 estrous cycles included in Experiment 1, cervical mucus samples collected before and after the treatment (on Day 3) were examined microscopically. The results were shown in Fig. 1. For several days after the treatment, the cervical mucus was cloudy and a large number of neutrophile leucocytes and epithelial cells and a small number of eosinophille leucocytes were observed in almost all of 13 animals in which estrous cycles were shortened after the treatment.

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The anti-PMSG serum were reacted to PMSG preparation and gonadotropin in pregnant mare serum. The reaction with anti-PMSG serum to HCG, FSH, HGH, ACTH and various animal sera were observed to be all negative. Immunological activity in sera of 264 pregnant mare was titer 4. The false negative cases of appearance of immunological pregnancy test were directly related to the concentration of the gonadotropin in pregnant mare serum.
The in vivo neutralization of PMSG by anti-PMSG serum was demonstrated. The materials to test the ability of antibody to inactivate PMSG were injected into 21-28 days immature female mice. No corpora lutea were observed in the fraction from ether solution, 100 IU/ml of PMSG in saline with anti-PMSG serum, precipitate containing the hormone-antibody complex and pregnant mare serum with anti-PMSG serum. While 100 IU/ml of PMSG in saline with normal rabbit serum, 100 IU/ml of PMSG in saline and pregnaut mare serum were showed prominent luteinization.

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[in Japanese]

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