The Effect of Genistein on the Plasma Membrane Integrity of Frozen Ongole Grade Bull Semen Based on Skim Milk – Soy Lecithin Extender

Abstract

Beef cattle represent livestock commodities that can help support national economic stability. Ongole grade bull (PO) is one of the local cattle breeds whose population and genetic quality decrease. Various efforts have been made in improving the genetic quality and in increasing the population of the local livestock population to achieve self-sufficient status of the beef supply in Indonesia. One of the efforts is artificial insemination (IB). The study aims at examining the effect of different concentrations of genistein semen extender on the post-thawing membrane integrity of Ongole grade bull semen. It is useful in developing the genistein as a protective agent against reactive oxygen species (ROS) during cryopreservation process. It was conducted at BPBPTDK, Center for Animal Breeding Development, Animal Feed, and Veterinary Diagnostics, in the Special Region of Yogyakarta from February 7th, 2019 to May 7th, 2019. The semen of two healthy Ongole grade bulls was consecutively collected five times in the period of 3 months using artificial vagina. The semen was cryopreserved using skim milk-soy lecithin semen based extenders at different concentrations of genistein, which were group A (1.0 mmol genistein), B (2.0 mmol genistein) and K (without any genistein as control). The observed parameters included individual spermatozoa motility, percentage of spermatozoa viability that was observed using Eosin nigrosine staining method and spermatozoa membrane integrity that was observed using the hypo osmotic swelling test (HOST) method. The observations were made on fresh semen, extended semen, and post-thawing semen. The results of the observation were then analyzed descriptively and statistically using regression correlation test and one-way analysis of variance (ANOVA). The results of the analysis showed that the addition of Genistein at the different concentrations of 1 mmol and 2 mmol resulted in higher motility, viability, and membrane integrity than the control and had significant effect (p<0.05) on the motility, the membrane integrity, and the acrosome integrity of the Ongole grade bull semen. The genistein concentration of 2.0 mmol gave the highest mean percentage of spermatozoa motility (57.50 ± 1.00 %) after freezing. The genistein concentration of 1.0 mmol gave the highest mean percentage of spermatozoa membrane integrity and acrosome integrity (52.45 ± 0.85 % and 49.58 ± 0.98 %) after freezing.