The purpose of this study was to determine the effect offertilization time and dffirent incubation systems onfertilization level by in-vitro. The mature of ovaries from indigenous cow andfresh cementfrom HolsteinFrisiancows(FH),0.9% NaClphysiological PBS,NissuiJapan, I,HEPES30pM,20 mL Heparin, 10% goat serum, 250 g/ml FSH BO medium, medium gentamisain 50 mB-O, mineral oil, alcohol, aquabidest, and I %o aceto orcein were materials and reagents. The Completely Randomized Design (CRD) in factorial pattern was used. Results shown no significant ffict on the percentage of fertitized ooq)tes and level of development of pronuclei (2PN and> 2PN) by different timing offertilization and incubation system. The development of pronucleus (IPN) showed significant (P <0.05) on I2 hours (37.60 %o), but no significant effect on dffirent incubation system. It concluded, the system of incubation and time of fertilization has no effect on oocyte fertilization rate. Oocytes fertilization time can be performed at 6 hours , I 2 hours, and I B hours, while the extension of the period of fertilization until I 8 hours did not increase the I ev el offertilization.
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