Artificial insemination with fresh semen in beagle bitches.

Tanaka

Takagi

et al. 2000

Self Cite

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.

Section: Discussionsupporting

confidence: 54%

“…The semen volume for AI was adjusted to 30 µl with egg yolk Tris-fructose citrate solution. IUI was performed as we previously reported for dogs [11,12]. After confirming the numbers of mature follicles and ovulation from each left and right ovary, a greater amount of semen was inseminated into the uterine horn.…”

Section: Animalsmentioning

confidence: 99%

Unilateral Intrauterine Horn Insemination of Fresh Semen in Cats.

Tanaka

Takagi

et al. 2000

Self Cite

“…As we previously reported [19,23], bitches were made to stand on their forelegs, and the semen was inseminated into the deep vagina. The bitches were kept in this position for 15 min.…”

Section: Animalsmentioning

confidence: 99%

“…We also preformed IUI using an extender containing no OEP in a small number of animals. IUI and IVI was performed referring to our results previously obtained using fresh semen [19,22,23]. …”

mentioning

confidence: 99%

Intrauterine and Intravaginal Insemination with Frozen Canine Semen Using an Extender Consisting of Orvus ES Paste-Supplemented Egg Yolk Tris-Fructose Citrate

Hase

Tanaka

et al. 2000

Self Cite

ABSTRACT. Our previous report indicated that addition of Orvus ES Paste (OEP) to the extender of frozen canine semen protected acrosomes and maintained sperm motility after thawing. In this study, artificial insemination (AI) using the frozen semen was carried out. The frozen semen was prepared using egg yolk Tris-fructose citrate, and the final concentrations of glycerol and OEP were 7% (v/v) and 0.75% (v/v), respectively. AI was performed during the optimal mating period predicted from the peripheral plasma progesterone level. In intrauterine insemination (IUI), the bitches were laparotomized and 1 × 10 8 spermatozoa were infused into one of the uterine horns. In insemination of non-OEP supplemented semen, 3 × 10 8 spermatozoa were inseminated. In intravaginal insemination (IVI), 10-40 × 10 8 spermatozoa were inseminated. Conception was obtained in nine of 10 bitches (90.0%) that underwent IUI. The number of newborns was from 1 to 7 (mean 3.6 ± 0.9). The mean ratio of the number of puppies to the number of ovulations in the inseminated uterine horn was 71.8%. The number of puppies did not exceed the number of ovulation in the inseminated uterine horn. Conception using non-OEP supplemented frozen semen was unsuccessful in all four bitches. In IVI, conception was not obtained in any of the six bitches that received insemination of 10 × 10 8 or 40 × 10 8 spermatozoa, but two of three bitches that received insemination of 20 × 10 8 spermatozoa were fertilized. It was shown that a high conception rate can be obtained by IUI using OEP-supplemented frozen canine semen. Developmenmt of a non-surgical method of IUI and a method of freezing canine sperm applicable to IVI is necessary.-KEY WORDS: artificial insemination, canine, frozen semen, Orvus ES Paste.Conception using frozen canine semen was initially achieved by Seager and Platz using pellet form semen in 1969 [14]. Following this, conceptions by the straw method were reported by Andersen in 1972 [1]. Thereafter, conceptions using frozen semen have been obtained at a small number of research institutions, using egg yolk Trisfructose citrate (EYT-FC) [1-4, 11-13, 17, 18, 24], Triladyl [8,10], laiciphos [15,16], biciphos [16], and CLONE [17] as extenders. The composition of some of these extenders has not been revealed. With regard to the insemination site, both intravaginal [2,4,[8][9][10][11][12][13]15] and intrauterine [3,4,6,12,13,15,16,24] insemination (IVI, IUI) have been performed. Moreover, the numbers of sperm and modes of insemination vary among researchers. A high conception rate has been documented in many of these reports. However, it is generally recognized that currently frozen canine semen is not practical for clinical use, contradicting the research reports.We have previously reported [20] that the quality of frozen semen prepared using EYT-FC, which is used as an extender of frozen canine semen, is reasonable, however, the sperm motility rapidly decreased in the early phase after thawing, and was reduced to 0% after three hours. Referring to the ...

“…The semen samples, collected by digital manipulation, were divided into three fractions. The volume of semen, the number, motility, and viability of sperm were determined by methods described previously [19]. The motility of spermatozoa was represented by the percent ratio of spermatozoa that actively moved forward using a test plate for semen property examination (FA225, Fujihira Industry, Co., Ltd., Japan).…”

mentioning

confidence: 99%

Effects of Orvus ES Paste on Canine Spermatozoal Longevity after Freezing and Thawing

Hase

Hori

et al. 2000

Self Cite

ABSTRACT. Because of weak resistance of canine sperm to freezing, an applicable method of preparing canine frozen semen has not yet been established. We added various concentrations of Orvus ES Paste (OEP) to egg yolk Tris-fructose citrate, and investigated its effectiveness on survival of spermatozoa. Addition of 0.5-1.0% OEP to the extender for freezing canine semen was effective in prolonging post-thaw survival of spermatozoa.-KEY WORDS: canine, frozen semen, Orvus ES Paste.The first successful fertilization using frozen canine semen involved semen pellets, and was achieved by Seager and Platz in 1969 [15]. Then, in 1972, Andersen reported a successful case by the straw method, which is convenient for storage [1]. Thereafter, many researchers have investigated extenders [5,7,8,17,18], glycerol concentrations [2,4,5,10,12,16], freezing methods [2,10,[16][17][18], and thawing methods [4, 5, 7, 10-12, 17, 20] for frozen canine semen, but an applicable method of preparing frozen canine semen has not yet been established. It is recognized that canine sperm have poor freezing resistance, and sperm motility cannot be maintained for a prolonged time after thawing [3,7,13]. This is considered to be due to major injury of the acrosomes during thawing.A surfactant, Orvus ES Paste (OEP, Nova Chemical Sales, Inc., MA, U.S.A.), is added to the extender of boar sperm, because of weak freezing-resistance. OEP is known to protect the sperm acrosomes after thawing, resulting in improvement of sperm motility and conception rate [6]. In 1993, Nöthling and Volkmann [9] succeeded in obtaining conception using canine frozen semen diluted with an extender supplemented with 0.5% OEP. In 1997 and 1999, Rota et al. [13, 14] also showed the effectiveness of addition of 0.5% OEP to the extender for canine frozen semen by observing the sperm motility and acrosomes after thawing.Although Nöthling and Volkmann [9] and Rota et al. [13,14] set the OEP concentration to 0.5%, the criterion for determining this concentration is not clear. Therefore, in this study, using Tris-fructose-citrate extender containing 20% (v/v) egg yolk (EYT-FC), as used by Rota et al. [13,14], we investigated the effect of concentration of OEP supplemented to the extender on post-thaw survival of canine spermatozoa. In previous reports dealing with frozen canine semen [2, 4, 8-11, 13, 14, 16, 17, 20], 0.5 ml volume straws were used. Since intravaginal insemination of frozen semen in dogs requires a large number of surviving spermatozoa, we investigated the possibility of semen preservation in 1.0 ml straws .The male dogs used in this study were 10 beagles from 2 to 6 years of age with copulation ability and fertility. The semen samples, collected by digital manipulation, were divided into three fractions. The volume of semen, the number, motility, and viability of sperm were determined by methods described previously [19]. The motility of spermatozoa was represented by the percent ratio of spermatozoa that actively moved forward using a test plate for semen prope...