Penentuan Waktu Optimal Pengujian Keutuhan Membran Plasma Spermatozoa Semen Beku Sapi Limousin dan Frisien Holstein Menggunakan Hypo-Osmotic Swelling (HOS) Test.

Abstract

The study was conducted to determine the optimal time to test the integrity of plasma membrane of Limousin and Frisien Holstein (FH) bull frozen semen spermpatozoa using hypoosmotic swelling (HOS) test. A total of 15 samples from each bull used. All samples were incubated in a hypoosmotic solution with osmolarity of 150 mOsm Kg-1 for 60 minutes. The hypoosmotic solution made by admix 1.351 g fructose, 0.735 Na-citrate and water until the volume was 100 ml. Every 15 minutes the whole sample was evaluated the percentage of spermatozoa that reacts to the solution. Optimal time was obtained at 30’ and 45’, both on Limousin and FH frozen semen. The mean percentage of reacted Limousin bull spermatozoa at 30’ and 45’ respectively were 53.90 ± 6.81 % and 52.47 ± 8.00 %. Whereas the FH bull spermatozoa was 44.52 ± 6.26 % and 44.46 ± 8.42 %. Spermatozoa from frozoen semen needs time to react to hypoosmotic solution due to the added of freezing diluent. This causes the spermaozoa did not react immediately to the hypoosmotic solution. Plasma membrane integrity is very important for sperm fertility. Fertility enzymes contained in the head of spermatozoa, thus if the plasma membrane of the head broken, spermatozoa will lose their fertility. Similarly in the tail, if the plasma membrane broken, spermatozoa will lose their motility.