การคัดแยกเพศจากอสุจิของแพะด้วยการปั่นเหวี่ยงผ่านชั้นต่างระดับของสารละลายเพอร์คอลล์

Abstract

The objectives of this study were to investigate the post-thawed semen quality and sex ratio of kids with sperm sexed by Percoll density gradient centrifugation in goat. The study was divided into 2 experiments which were: Experiment 1: effect of Percoll density gradient centrifugation on post-thawed sperm quality The semen was collected from three mature Boer bucks, 3-5 years of age, once a week for 3 consecutive weeks using artificial vaginal method. Each week, the semen with progressive motility and concentration greater than 70% and 2,000 million sperm per milliliter, respectively, were pooled. The pooled semen was then divided into 2 groups of with or without centrifugation before frozen semen processing. For the centrifugation group, the semen was centrifuged using 7 layer of Percoll density gradient (40, 45, 50, 65, 70, 80 and 90%). After centrifugation, only semen from 2 layers of 80 and 90% Percoll were further processed for frozen semen. *Post-thawed semen quality was analyzed by CASA at 0, 1, 2, 3, 4, 5 and 6 h after incubation at 37°C. The results showed that the interaction between [treatment (with or without centrifugation) and times was highly significant difference (P<0.01) for sperm motility, whereas the progressive motility, velocity (VAP, VSL and VCL) and kinetic movement (ALH, BCF, STR and LIN) of sperm were not significant (P>0.05). In addition, the centrifugation using Percoll density gradient did not effect on progressive motility, velocity and kinetic movement of sperm. From this study it could be concluded that the centrifugation using Percoll density gradient had no effect on post-thawed semen quality in goat. In addition, Percoll density gradient technique could be applied for goat sperm sexing. The benefit of this study could be used for frozen sexing sperm production, using percall density gradient centrifugation, in goat. Experiment 2 effect of insemination with sexed sperm by Percall density gradient centrifugation on sex ratio af kids A total of 90 does (first-parity or greater) were used in this study. The body weight af does was between 25-30 kilagrams. Does were in 3-35 body condition score (score 1-5). The experimental does were divided into 2 gaugs af control (m = 46) and treatment (in = 44). The does in contral graup were inseminated with frozen unsexed sperm whereas in the treatment group were inseminated with frozen sexed sperm by Percoll density gradient centrifugation. The does were inseminated with frozen-thawed semen (150 million sperm per dose) at 12=24 m alter the anset of estrus ance or twice (12 h interval). Mostly does (n=75) were recevet one insemination and another 15 doses were NO at 12 h interval. Concagtion was determined by trans-rectal ultrasonography at about 45-40 days after insemination and the sex of kids were detected at kidding The results showed that the conception rate of does was 78.26 and 70.45 % in control and experimental group, respectively. The sex rattio of kids was significantly different between two graugs (Pcd:0172). The percentage di female and male kids in contrall group was 70.45 and 29.55, respectively, whereas in treatment group was 45 65 and 54.35,, respectively. Im conclusion, the Fercail density gracient centrifugation could be used for sperm sexing in goat to increase female kid