ABSTRACT
Background and Aim: Egg yolk (EY) is commonly used as an extracellular cryoprotectant in semen diluents but has some negative effects. Therefore, this study aimed to assess the potential of lecithin derived from plants, such as soybeans, as an alternative extracellular cryoprotectant and to characterize liquid semen quality of Ongole crossbred bulls using a modified caudal epididymis plasma-3 [CEP-3 (m)] as a base diluent and aqueous soybean extract (ASE). Materials and Methods: A bull with progressive motility (PM) of fresh semen >70% was used. Two soybean extracts were also used, namely, ASE 1 and ASE 2, obtained by extraction procedures 1 and 2, respectively. The study was conducted using an experimental design with 11 treatments and ten replications, with diluents comprising different levels of ASE 1 and ASE 2, as well as a positive control with 10% EY. The parameters measured were motility (M) and its kinetic parameters, including PM, M, velocity curve linear, velocity straight linear, velocity average pathway, linearity, straightness, wobble, amplitude lateral head beat cross frequency, and hyperactivity using computer-assisted sperm analysis, viability, and spermatozoa abnormalities. Results: The CEP-3(m) diluent formula and ASE 1 at a 30% level maintained the PM of spermatozoa up to day 5 (40.7% ± 16.1%) of cold storage. Meanwhile, the CEP-3(m) diluent formula and ASE 2 could only maintain PM >40% until day 3 (42.1% ± 13.5%) of cold storage at a 30% level. The CEP-3(m) diluent and ASE 1 at a level of 25%-30% supported spermatozoa life (viability) up to day 5 with a value >80% (81.8 ± 3.5; 86.4 ± 2.6). The abnormality value of spermatozoa in various diluents during cold storage on days 0-5 was below 20%. Conclusion: Soybean extracts 1 and 2 can substitute EYs as extracellular cryoprotectants in modified CEP-3 basic diluents. Soybean extract 1 can support the life of spermatozoa up to day 5 but may cause the viscosity and movement of spermatozoa to be hyperactive. Soybean extract 2 can support the life of spermatozoa up to the 3rd day of cold storage and produces progressive (non-rotating) movement patterns. Further, research is recommended with higher levels of ASE 2.
ABSTRACT
BACKGROUND AND AIM: Lecithin based diluent such as AndroMed, is a semen diluent made without animal components to prevent the risk of disease transmission, while glutathione (GSH) is an intracellular non-enzymatic antioxidant that prevents cell damage due to reactive oxygen species. Meanwhile, the specific impact of AndroMed and GSH combinations on spermatozoa abnormalities has not been fully studied. Therefore, this study aims to determine the effect of using cauda epididymal plasma-2 (CEP-2) and AndroMed diluents with or without the addition of GSH on the abnormalities of sexing semen of Ongole crossbred bulls in cold storage. MATERIALS AND METHODS: This study used a factorial completely randomized design 2×2, the first factor was types of diluent and the second was with or without the addition of GSH. Observation of spermatozoa abnormalities was carried out at a storage time of 0-5 days using 297 ejaculations of liquid semen, with 100 spermatozoa observed per smear of each ejaculate. Furthermore, the data were analyzed using a two-way analysis of variance and the significant threshold (p-value) for statistical analysis was set at <0.05. RESULTS: The results showed that there was a significant difference (p<0.05) between AndroMed and CEP-2 in minimizing the abnormalities of upper layer spermatozoa (X), with parameters DH and AD on day 0, damage of spermatozoa (DMR) on days 1-5, and dag-like defect (DLD) on day 5. Furthermore, spermatozoa abnormalities in the lower layer (Y) showed a significant difference between diluents in the parameters of AD on day 1, DMR on days 0-5, and DLD on days 1-5. The significant difference between with or without the addition of GSH in the X sperm was observed in the DH parameters on day 0 and DMR on 5, while there was no significant difference in the Y sperm. CONCLUSION: Based on the results, AndroMed has the potential to minimize spermatozoa abnormalities compared to CEP-2 diluent in sexed liquid semen. Therefore, AndroMed diluents with or without the addition of 1 mM GSH have no significant effect on spermatozoa abnormalities.
