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

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 × 108 spermatozoa were infused into one of the uterine horns. In insemination of non-OEP supplemented semen, 3 × 108 spermatozoa were inseminated. In intravaginal insemination (IVI), 10-40 × 108 spermatozoa were inseminated. Conception was obtained in nine of 10 bitches (90.0%) that underwent IUI. The number of newboms 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 × 108 or 40 × 108 spermatozoa, but two of three bitches that received insemination of 20 × 108 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.