Seleksi Spermatozoa Domba Garut dengan Metode Sentrifugasi Gradien Densitas Percoll
Abstract
Good quality of sperm was necessary prepared in order to increase the efficacy of in vitro fertilization. The good quality of sperm could be obtained through selection using percoll density gradient centrifugation method. The aim of this research was to determine an optimal combination of speed and centrifugation time in providing better quality of sperm with the lowest mortality rate or damage. Garut sheep fresh semen with 200 million cells/ml in concentration was used as research sample for each treatments. Factorial completely randomized design was used as research design, with one control and sixteen treatments combination of centrifugation speed and time: (1) 200 xg for 5 min; (2) 200 xg for 10 min; (3) 200 xg for 15 min; (4) 200 xg for 20 min; (5) 300 xg for 5 min; (6) 300 xg for 10 min; (7) 300 xg for 15 min; (8) 300 xg for 20 min; (9) 400 xg for 5 min; (10) 400 xg for 10 min; (11) 400 xg for 15 min; (12) 400 xg for 20 min; (13) 500 xg for 5 min; (14) 500 xg for 10 min; (15) 500 xg for 15 min; and (16) 500 xg for 20 min. All of control and treatments combination were replicated for three times. The sperms variables were observed sperm concentration, percentage of live sperms, percentage of progressive motility sperms and percentage of normal sperms. Data were evaluated by two way analysis of variance (ANOVA), and differences between treatment group were determined by a post hoc, Duncan’s multiple range test (DMRT). Differences were considered significantly at a probability value (P) of 0.05. The results showed that the treatment of centrifugation 400 xg for 15 min gave the optimal combination significantly (P<0.05) with the highest sperms concentration (33.33%), the highest percentage of living sperms (74.82%), the highest percentage of progressive motility sperms (57.56%), and the highest percentage of normal sperms (86%) were obtained. It is concluded that sperms selection using percoll density gradient centrifugation method is able to increase the percentage of sperms concentration, percentage of live sperms, percentage of progressive motility sperms, and percentage of normal sperms. Centrifugation should be done at the speed of 400 xg for 15 min to decrease the damage rate of sperms.