Influence of the type of semen and morphology of individual sperm cells on the results of ICSI in domestic cats.

The aim of this study was to analyze the influence of the type of spermatozoa and of different sperm abnormalities on fertilization and embryo development after ICSI in cats. In Exp I, ICSI was performed using urethral or epididymal spermatozoa collected from 7 tomcats. In Exp. II, epididymal spermatozoa from 16 cats were used for ICSI and an epididymal spermatozoon exhibiting no abnormalities or one with an abnormality was microinjected into an oocyte. Exp. I was performed in 14 replicates and Exp. II was performed in 20 replicates. In both experiments the number of cleaved oocytes, the number of embryos at the morula stage and the number of embryos at the blastocyst stage were evaluated at 24 h, and at 6 and 7 days after ICSI, respectively, and compared between experimental groups. No statistically significant differences (P > 0.05) were observed, either for Exp. I or for Exp. II. The average cleavage rate was 60.2%, morula rate 62.3% and blastocyst rate 19.2% in Exp. I and 51.6%, 66.8% and 25.8% in Exp. II, respectively. The study confirmed that both urethral and epididymal spermatozoa can be used for in vitro fertilization in cats and proved the usefulness of the ICSI method in the case of teratozoospermic males. The study showed that even in severe cases, when almost no normal spermatozoa can be found in the semen, it is possible to obtain embryos using abnormal sperm cells with the same chance of success as for normal spermatozoa.

The use of human and bovine commercial media for oocyte maturation and embryo development in the domestic cat (Felis catus).

The aim of this study was to examine the suitability of commercial media designed for humans and cattle for oocyte maturation and embryo culture in the domestic cat. In Exp. I, feline oocytes collected ex vivo were subjected to in vitro maturation in a laboratory-made culture medium (based on M199) or a commercial medium designed for cattle cells (BO-IVM® ). In Exp. II, ICSI-derived feline embryos were cultured for 7 days in a commercial human (Continuous Single Culture® ) or bovine (BO-EC® ) cell medium. The rates of cleavage, morula and blastocyst formation were evaluated at 24 hr, 6 days and 7 days after ICSI, respectively, and compared between experimental groups. At the end of culture, embryos were assessed for viability and apoptotic changes. In Exp. I, no statistically significant difference in oocyte maturation outcome between laboratory-made (52.7%) and commercial media (58.9%) was observed. However, the use of a commercial medium prepared for use with bovine cells resulted in a significantly lower variance of the maturation rate. In Exp. II, no statistically significant differences between two commercial media were observed for cleavage (67.5% and 64.5%), morula (39.3% and 47.1%) and blastocyst rates (25.0% and 19.6%), as well as for the percentage of late apoptotic blastomeres. Morulae cultured in medium marketed for humans exhibited significantly more early apoptotic (43.2 ± 31.2% vs. 23.4 ± 23.2%) and necrotic (60.6 ± 47.6% vs. 29.4 ± 22.6%) blastomeres. In conclusion, both commercial media tested are suitable for in vitro oocyte maturation and embryo culture procedures in cats. It is remarkable that a culture medium designed for use in cattle for in vitro maturation of cat oocytes provides more reproducible results.