Всероссийский научно-исследовательский институт физиологии, биохимии и питания животных – филиал Федерального государственного бюджетного научного учреждения «Федеральный научный центр животноводства – ВИЖ имени академика Л.К. Эрнста»
ABSTRACT. The stage of in vitro fertilization (IVF) is considered as a pivotal term in the embryo production programs in mammals, although the effect of this stage is still underestimated, especially in sheep. In IVF laboratories, oocytes are exposed to spermatozoa for 18 to 24 h. This prolonged duration of co-incubation for gamete can produce high levels of reactive oxygen species (ROS), which can potentially cause an increase in polyspermic penetration and hardening of the zona pellucida (ZP), which may affect embryo viability. Despite this, most researchers still prefer 18-24 h of fertilizing time for in vitro ewe oocytes. This study aimed to improve the success rate of the IVF process in sheep by manipulating the time of the IVF stage. In addition, penetration time of ram spermatozoa into the oocytes was measured in vitro condition. For this purpose, maturated oocytes were incubated with capacitated spermatozoa at a concentration of 10×106 spermatozoa/ml in BO medium for either 6 h (Group 1, n = 119), 12 h (Group 2, n = 112), or 18 h (Group 3, n = 107). Our study showed that exposing gametes to IVF for only 6 h results in a higher fertilization rate and an improvement in the quality of the embryo development when compared to 12 h and 18-h co-incubation of gametes. Moreover, this study showed that sperm can arrive at the surface of zona pellucida already within 110 min after the co-incubation beginning and then, fertilization happens within 4.5 h after having started IVF procedure. Concluded that using the protocol adopted in this study, the highest percentage of zygote development to the blastocyst stage occurred in the shortest IVF period. In conclusion, the highest percentage of zygote development into the blastocysts stage occurs in the shortest period of IVF (6 h)
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