ABSTRACT. Artificial insemination with frozen cauda epididymal sperm was performed in cats. Sperm were transmigrated from the epididymides in 10 male cats. The mean sperm motility and viability were 67% and 82.5%, respectively, and 11.6 × 10 7 sperm were recovered. The mean sperm motility after thawing was 24.0%. Eleven female cats received unilateral intrauterine insemination of 5 × 10 7 sperm, and the conception rate was 27.3% (3/11). This was the first case of conception obtained with frozen epididymal sperm in cats. KEY WORDS: artificial insemination, epididymal sperm, feline.J. Vet. Med. Sci. 65(3): 397-399, 2003 It has been demonstrated in many species that epididymal sperm have fertility in in vitro fertilization and artificial insemination (AI) [4,8,[11][12][13][14][15]25], and fertilization with frozen epididymal sperm was confirmed in pigs [11] and dogs [9].Freeze-storage of epididymal sperm is an important technique for gamete preservation of species, particularly for preservation of animals becoming extinct. As wild animals in the cat family are becoming extinct, we selected cats as an animal model.We have performed studies of reproductive techniques such as in vitro fertilization [5,6], embryo transfer [6,24], and AI [18,[20][21][22][23] in cats.With regard to feline epididymal sperm, studies of the qualities [1,2], state of fertility acquisition [2,3,12], and freeze-storage method [4,8,17] are progressing, and fertility of fresh epididymal sperm [12] and frozen semen [4,8] has been shown, but no in vivo study with frozen epididymal sperm has been performed. With frozen feline ejaculated semen, Platz et al.[16] and we [23] have succeeded in obtaining newborns by intravaginal and intrauterine insemination, respectively. Therefore, we froze epididymal sperm by the same method as for freezing ejaculated sperm and investigated the possibility of inducing conception by intrauterine insemination. It has been shown that frozen semen supplemented with Orvus ES Paste (OEP, Nova Chemical Sales Scituate, Inc., MA, U.S.A.) has superior properties after thawing semen without OEP in dogs [9] and pigs [7]. In this way, the usefulness of OEP was also investigated in this study.Animals: Ten male crossbred cats aged 1.0-3.3 years (mean: 1.9 ± 0.3, SE) were used. For AI, 11 female cats aged 2-4 years were used. The cats were kept in groups in an animal room measuring 4 × 7 m with temperature controlled to 23 ± 2°C. The animals were given commercial dry cat food (Hill's Feline Maintenance Dry, U.S.A.).Transmigration of sperm from epididymis: Surgically excised testis and epididymis were weighed. The epididymis was cut at the corpus into two equal parts. Each part was minced in 1 ml of egg yolk Tris-fructose citrate solution (EYT-FC) [23] to transmigrate the sperm. The solution was filtered through a stainless steel mesh (80 µm) and sperm were recovered. The time from excision of the epididymis to transmigration of the sperm was within 1 hr.Sperm quality test and freezing: The recovered sperm were examined by the gen...

ABSTRACT. The sperm count required were investigated to obtain a conception rate of 80% by unilateral intrauterine insemination (UIUI) of fresh semen in cats. The conception rates obtained by insemination before and after ovulation were also examined. Thirty-six female cats aged 1-7 years were used in the experiments, and the number of experimental cases was 44. Seven male cats aged 2-12 years from which semen could be collected by the artificial vagina method were used. In artificial insemination, 100 iu × 2 or 250 iu of hCG was administered on days 2-4 of estrus, and sperm were introduced into the uterine horn with a greater number of ovulations (or mature follicles) 15, 20 and 30 hr after hCG administration by laparotomy. The inseminated sperm counts were 2 × 10 6 (Exp. 1), 4 × 10 6 (Exp. 2), and 8 × 10 6 (Exp. 3). As a result, ovulation was induced in 42 of 44 cases (induction rate: 95.5%) regardless of the dosage of hCG. Conception was obtained by UIUI in two of 16 animals (conception rate: 12.5%) in the Exp. 1, five of 16 animals (31.3%) in Exp. 2, and eight of 10 animals (80.0%) in Exp. 3. Regarding the relationship between the ovulation state at insemination and conception, the conception rate obtained by insemination before ovulation was clearly higher than that obtained by insemination after ovulation (p<0.05). Regarding the number of kits compared to the number of ovulations on the inseminated side, the percentages of cases in which the number of kits exceeded the number of ovulations on the inseminated side were similar in all groups inseminated with a different number of sperm. It is therefore necessary to investigate conception rates obtained by bilateral insemination to increase the fertility rate. Based on the above findings, it was shown that the sperm count required for fertilization by UIUI is 8 × 10 6 . KEY WORDS: feline, fresh semen, intrauterine insemination.

ABSTRACT. Frozen feline semen was prepared using two types of extenders, egg yolk Tris-fructose citric acid (EYT-FC) and egg yolk sodium citrate solution (EYC), and the semen qualities after thawing and the conception rates obtained by unilateral intrauterine horn insemination (UIUI) were investigated. Cats used in the experiment were six males and 11 females aged 2-12 years (the number of experimental cases was 17). For preparation of frozen semen, semen collected by the artificial vagina method was adjusted to 1 × 10 8 sperm/ml and 7% glycerol, put in 250 µl straws, and then frozen using a cell freezer. The mean sperm motility after thawing was 30.0 ± 9.7 (SE) % in the semen prepared with EYT-FC and 30.0 ± 3.3% in the semen prepared with EYC. Four of seven animals were fertilized by UIUI using two straws in both extenders, and the conception rate was 57.1%. The mean ratios of number of kits to the number of ovulations in the inseminated side were 61.1 ± 24.5% and 30.5 ± 3.4% for EYT-FC and EYC, respectively, showing that the ratio tended to be higher in the semen prepared with EYT-FC. The above findings, comparing the two extenders for preparation of frozen feline semen, showed that EYT-FC is slightly superior to EYC. To increase conception and fertility rates, it may be important to increase the sperm count for insemination and to inseminate both uterine horns. KEY WORDS: feline, frozen semen, intrauterine insemination, straw method.J. Vet. Med. Sci. 62(12): 1247-1251 Feline artificial insemination (AI) using frozen semen has been reported only by Platz et al. in 1978 [3]. In their study, 50-100 × 10 6 viable sperms were inseminated into the vagina. Although only six of 56 animals were fertilized, obtaining a conception rate of 10.7%, these were the first cases of fertilization by frozen semen in cats. We previously performed intravaginal insemination (IVI) of fresh semen and clarified that a number of sperms, 80 × 10 6 , are required for fertilization [6]. This sperm count is equal to the number of sperm in two ejaculations in cats [5]. We also clarified the sperm count required for fertilization by unilateral intrauterine horn insemination (UIUI) of fresh semen to be 8 × 10 6 in cats, which is 1/10 of that by IVI [7]. Freezing markedly reduces motility and viability of sperms. Accordingly, it is important to develop a technique of fertilizaiton using a small number of sperm. Therefore, intrauterine insemination (IUI) may be appropriate for feline AI using frozen semen rather than IVI.The method using frozen feline semen reported by Platz et al. was the pellet method, which may pose difficulties for storage and use [3]. Therefore, we investigated freezing of feline semen by the straw method.Platz et al. used egg yolk lactose as the extender for frozen feline semen [3]. However, the semen qualities after thawing were not reported, and thus, we could not evaluate the extender used in their study. Therefore, we prepared frozen feline semen using egg yolk fructose citric acid (EYT-FC) [6,7], which we use as an e...

subjective evaluations at 9 stations in Tokyo under low illuminance condition by power saving and under usual condition, after The Great East Japan Earthquake.As the result, it shows that users perceive the changes of illuminance between two conditions, and approve low illuminance conditions.And it is suggested that the evaluation about feelings is less influenced by the change of illuminance than the evaluation about brightness, and is influenced by other factors such as a form of lighting and a spatial form. subjective evaluations at 9 stations in Tokyo under low illuminance condition by power saving and under usual condition, after The Great East Japan Earthquake. Keywords : Station, Lighting, Power Saving, User Evaluation, Spatial FormAs the result, it shows that users perceive the changes of illuminance between two conditions, and approve low illuminance conditions.And it is suggested that the evaluation about feelings is less influenced by the change of illuminance than the evaluation about brightness, and is influenced by other factors such as a form of lighting and a spatial form. As the result, it shows that users perceive the changes of illuminance between two conditions, and approve low illuminance conditions.And it is suggested that the evaluation about feelings is less influenced by the change of illuminance than the evaluation about brightness, and is influenced by other factors such as a form of lighting and a spatial form. subjective evaluations at 9 stations in Tokyo under low illuminance condition by power saving and under usual condition, after The Great East Japan Earthquake. Keywords : Station, Lighting, Power Saving, User Evaluation, Spatial FormAs the result, it shows that users perceive the changes of illuminance between two conditions, and approve low illuminance conditions.And it is suggested that the evaluation about feelings is less influenced by the change of illuminance than the evaluation about brightness, and is influenced by other factors such as a form of lighting and a spatial form.