The effect of arbutin on freezing of ram sperm

This study was conducted to determine the effects of arbutin, an antioxidant added to the sperm diluent after thawing, on spermological parameters, Computer-Assisted Sperm Analysis (CASA), oxidative stress parameters, and DNA damage. Sperm samples were collected from a total of 3 male goats of the Sönmez breed raised under Afyonkarahisar conditions. The samples were pooled and divided into 5 equal volumes, which were then diluted with Tris-based extenders containing different amounts of arbutin, including the control group. The diluted samples were subjected to equilibration at 5 °C for 3 hours in 0.25 ml straws before being frozen in liquid nitrogen vapor. Subsequently, the sperm samples were stored in a liquid nitrogen tank (-196°C) until the day of analysis. In terms of subjective motility and CASA motility, the group containing 2,5 mM arbutin in the thawed sperm showed a significant superiority compared to both the control group and the other groups. The parameters VAP, VSL, VCL, ALH, and BCF exhibited the highest values in the group containing 10 mM arbutin. Regarding the midpiece abnormalities of the spermatozoa, a significant decrease was observed in the 7,5 mM and 10 mM groups compared to the other groups and the control group (p<0.05). When looking at tail abnormalities, the lowest value was found in the 2,5 mM group, and the difference between this group and the 10 mM group was significant (p<0.05). Among the HE test parameters of the spermatozoa, the lowest membrane integrity and viability (H+, E-) were observed in the control group, while the highest value was found in the 7,5 mM group, and the difference between them was statistically significant (p<0.05). The values obtained for DNA damage in the groups containing 2,5; 5, and 7,5 mM arbutin were statistically significant (p<0.05) compared to the control group. Among the oxidative stress parameters, the values of MDA (malondialdehyde) in the 2,5 and 5 mM groups were significantly lower (p<0.05) compared to the 10 mM group. In the GSH (glutathione) parameter, the value obtained in the 10 mM group was statistically different (p<0.05) from the control and other groups. The highest value obtained in the total antioxidant level showed a significant difference (p<0.05) with the 7,5 and 10 mM groups. In terms of total oxidant level and oxidative stress index, a statistically significant difference (p<0.05) was observed between the 10 mM group and the control and other groups. In conclusion, it has been observed that arbutin, used as an antioxidant in the freezing of goat sperm, has positive effects on motility, CASA parameters, abnormal spermatozoa rate, membrane integrity, oxidative stress, and DNA damage. However, it is deemed beneficial to evaluate its effects along with fertility results using a wider range of animal samples.