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This study aimed to investigate the fertilization rate of oocytes using sperm from the cauda epididymis of Bali cattle until cleavage. The research spanned 10 months, involving various stages conducted at different facilities. Testicular samples were obtained from the Cibinong Slaughterhouse, followed by freezing of epididymal sperm at the Reproductive Rehabilitation Unit laboratory of the Faculty of Veterinary Medicine, Bogor Agricultural Institute. Post-thaw analysis of frozen sperm was performed at the Reproduction, Breeding, and Animal Cell Culture facilities of the LIPI Cibinong Biotechnology Research Center. At the same time, the in vitro fertilization (IVF) process took place at the Cipelang Bogor Livestock Embryo Center. The primary objective of this investigation was to assess the fertility of cauda epididymal sperm post-thawing, specifically evaluating the impact of caffeine supplementation on the success of in vitro fertilization (IVF). The methodology involved collecting cauda epididymal sperm at the slaughterhouse, with subsequent analysis of sperm quality after thawing by adding different concentrations of caffeine (T0: 0 mg/ml, T2: 2 mg/ml, T4: 4 mg/ml, T6: 6 mg/ml) in four replicates. The key parameter examined was the fertility of cauda epididymal sperm using the IVF technique. The experimental design utilized a one-way Completely Randomized Design (CRD), followed by Tukey-W-Procedure testing using SPSS 16. The findings revealed that the fertility of cauda epididymal sperm at concentrations T2 and T4 was significantly higher (P<0.05) compared to T0 and T6. Consequently, the study concluded that the addition of 4 mg/ml caffeine to thawed cauda epididymal sperm resulted in a 37.50% success rate in the IVF process and a 10% cleavage rate of oocytes in Bali cattle.
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