To induce luteal regression-related abortion/delivery and treat pyometra in dogs, various PGF2α-analogues (PGAs) are administered, but a PGA most appropriate for clinical application in dogs, with a low incidence of side effects, is being investigated. In this study, we compared the effects of etiproston tromethamine (PGA-E), which has not been investigated in dogs, with those of cloprostenol (PGA-C), which is routinely used in dogs. A single dose of PGA-E at 100, 200, 400 or 800 μg or PGA-C at 12.5, 25, 50 or 100 μg was administered to beagles (n=5 per group) 25 days after ovulation, when the corpus luteum was in the functional phase. We compared the state of luteal regression by measuring plasma progesterone levels. As side effects, the incidences of salivation, vomiting, tachypnea, diarrhea and the drop in body temperature were investigated. In the 400-μg and 800-μg groups treated with PGA-E, the mean intervals from administration until luteal regression were 18.6 days and 31.2 days, respectively. In the dogs treated with 50 μg or more of PGA-C, luteal regression was noted 2 days after administration. The above side effects were observed for 3 hr after administration of PGA-E/PGA-C. In the dogs treated with 800 μg of PGA-E, the mean body temperature was 36.7°C 4 hr after administration; hypothermia persisted. PGA-E may be less useful than PGA-C for promoting luteal regression in dogs in clinical application.
A Beagle with a low plasma testosterone (T) level and azoospermia was given 10 subcutaneous injections of 1 μg gonadotropin-releasing hormone analogue (GnRH-A) per head at intervals of 3 days (Experiment 1), and 6 months after the final injection was given, 15 subcutaneous injections of 2 μg GnRH-A were given at intervals of 2 days (Experiment 2). The plasma T level increased and peaked at 8 weeks after the first injection of GnRH-A in both Experiment 1 and Experiment 2. Motile sperm were detected in the semen collected 8 weeks and 7 weeks after the first injection in Experiment 1 and Experiment 2, respectively. The total number of sperm peaked 9 weeks after the first injection in both Experiment 1 (4.5 × 106) and Experiment 2 (72.8 × 106).
In dogs, embryo transfer (ET) techniques such as induciton of excessive ovulation and synchronization of estrus have not progressed well. Therefore, using embryos at various developmental stages, ET was investigated in dogs from a beagle colony in which the ovulation days were close, as estimated by the progesterone level. Embryos were recovered 8-11 days after ovulation (4-9 days after mating) by excising the oviducts and uteri (excision method) in 16 animals and by surgical flushing of the uteri at laparotomy (surgical method) in 3 animals. In 24 dogs with -4 to +2 days of difference in the timing of ovulation between donor and recipient dogs, 1-10 embryos at the 8-cell to blastocyst stages were transferred per animal. The mean embryo recovery rate by the excision method (97.1%) was significantly higher than that by the surgical method (42.5%) (p<0.01). Twelve (57.1%) of 21 animals with -1 to +2 days difference in ovulation day became pregnant after the transfer of 8-cell to blastocyst stage embryos. Although 3 dogs with -4 to -2 days of difference of ovulation day underwent ET of morula or compacted morula, none of these dogs became pregnant. The mean ratio of the number of newborns to the number of transferred embryos was only 51.9%. The mean duration of the period between ovulation and delivery in the pregnant recipients was 65.8 days, which tended to be longer than that in natural mating. These results demonstrate that pregnancy can be induced by ET at the 8-cell to blastocyst stage in dogs with -1 to +2 days difference in ovulation day.
The effects of protein supplements and culture dish type on in vitro fertilization (IVF) and embryo development in culture were examined in the domestic cat. In Experiment I, follicular oocytes were fertilized and cultured in either 1) modified Earle's balanced salt solution, designated MK-1, supplemented with one of the following: 10% human serum (HS), 10% FCS or 0.4% BSA, or 2) Medium 199 (M-199) supplemented with 10% FCS. Fertilization rates were lower (P<0.01) in MK-1 + BSA (74.4%), MK-1 + FCS (56.1%), and M-199 + FCS (51.4%) than in MK-1 + HS (94.7%). A greater (P<0.01) percentage of blastocysts was obtained in MK-1 + HS (50.0%) than in other treatment groups (range, 4.3-17.2%). In Experiment II, the effect of dish type (tissue culture dish, TCD, versus suspension culture dish, SCD) on embryo development was evaluated in MK-1 supplemented with either HS or BSA. Significantly higher proportions of IVF-derived embryos developed to blastocysts at 120 and 144 hr post-insemination, respectively, when cultured in HS/SCD (47.2 and 71.7%) than in BSA/SCD (11.4 and 27.3%) or BSA/TCD (10.4 and 25.0%). At 120 hr post-insemination, there was a lower (P<0.01) percentage of blastocysts in HS/TCD (22.2%) than in HS/SCD. In Experiment III, six embryos per cat were transferred to the uterine horns of 17 recipients at 144 hr after hCG treatment. Five of 7 recipients which received late morulae cultured in MK-1 + BSA (SCD) for 120 hr became pregnant (71.4%). Eight of 10 recipients which received early blastocysts cultured in MK-1 + HS (SCD) for 120 hr became pregnant (80.0%). We conclude that MK-1 containing HS is highly beneficial for overcoming the in vitro developmental block of IVF-derived feline embryos and increasing the success rate of IVF/ET.
The relationship between ejaculation intervals and semen quality in 4 male cats aged 3−5 years was investigated in this study.Semen was collected 10 times at intervals of every day, every other day, and every three days using an artificial vagina.Semen was collected consecutively twice on the day of semen collection, and the semen quality was examined.In semen collected every day, the number of sperm in the first collection decreased, and the frequency of immature sperm rapidly increased after the 4th day.In semen collected every other day and every three days, although the semen volume markedly varied among the animals on both first and second collections, the volume remained stable for each animal, the number of sperm was similar in the first and second collections, but was clearly larger in the first collection(p<0.01).Sperm motility and abnormality were stable among the various intervals and between the first and second collections in each animal.
We collected semen from a male Amur leopard cat using the transrectal electroejaculation method and investigated the semen qualities for about four years. In addition, the influence of the season on the spermatogenic function of the Amur leopard cat was investigated with regard to the semen qualities, testicular volume and serum testosterone level. As a result, we could collect semen with good sperm qualities that would be useable for artificial insemination. Some seasonality was noted in the testicular volume and serum testosterone level. We clarified that the semen qualities were favorable before and during the female breeding season compared with those after the breeding season.
The fertility was compared between ejaculated and cauda epididymal sperm sensitized with prostatic fluid in dog after freeze-thawing using the fertility of ova from the contralateral ovary after injection (2 × 108 sperm) into dog uterus on the unilateral ovariectomized side, on the basis of the presence or absence of conception. No significant difference was observed in sperm quality after freeze-thawing between the two groups and conception rates were equivalent and low. Therefore, to achieve a high fertility by intrauterine insemination of canine frozen-thawed ejaculated and cauda epididymal sperm, intrauterine insemination on both sides is recommended, rather than insemination with a lot of sperm of the uterine horn on one side.
