This study was conducted to investigate the optimal artificial insemination (AI) time with diagnostic kit at ovulation time. We already applied the patent about the protein in the cow heat mucose in external reproductive tract. And we would examine the accuracy for detection of cow heat by the kit produced with the protein. Evaluation of optimal heat detection was tried two time at 12 hrs and 24 hrs after the heat. And then, AI service also performed two times with no relation to the results of heat diagnosis by heat detection kit and pregnancy rates were checked with rectal palpation on 60th day after AI. Heat diagnostic results by kit in natural heat after 12 hrs in Hanwoo cows were showed 31.3~75.0% on positive in first heat detection and 33.3~100.0% on positve in second heat detection. In the 1st positive results were significant different (p<0.05), but 2nd positive were not. The results of heat detection showed different result on regional influence and individual cow effects. The pregnancy rates of first trial of heat detection were showed 34.4~78.7% on positive and 21.3~68.8% on negative after the diagnosis by heat detection kit. And the pregnancy rates of next trial of heat detection were showed 33.3~85.7% on positive and 14.3~66.6% on negative after the heat diagnosis. Both positive results of first trial and next trial also were showed significant different (p<0.05), but negative results were not. In positive result, first trial of total pregnancy rates was higher than the next trial of pregnancy, but there showed opposite results on negative results. In conclusion, the optimal heat detection kit is suitable to ordinary Hanwoo cows and it suggested that we have to improve the kit’s accuracy by detecting the materials like proteins related optimal AI time.

The boar sperm has more lipid droplets and specialty of seminal plasma compared with other species, causing difficulties of freezing sperm and decreases for the utilization of frozen semen into the artificial insemination. However, several studies reported significant results for the recovery of sperm motility and reproductive by addition of cryoprotectants and seminal plasma after thawing. This study was designed to investigate the effects of supplementation of trehalose or glycerol in the LEY (lactose and egg yolk in BTS) solution for the conventional freezing and vitrification process. Two boars aged 16 months were used to collect semen for 2 times in a week. The samples were allotted to 3 freezing solutions (LEY + glycerol 10.5% + OEP 1.5%, LEY + trehalose 1M + OEP 1.5%, and sucrose 1.5M + trehalose 1 M + OEP 1.5%) after centrifugation at 800 g for 10 minutes. Semen was equilibrated in freezing solutions for 10 minutes and injected into plastic straws with 2∼3 air bubbles to minimize freezing damages. Vitrification was performed to locate sperm in 5 cm above LN2 for 5 minutes, and the conventional freezing was conducted with an automatic freezer. Motility and survival rates were measured by CASA (Computer assisted sperm an alyzing system) and FITC (Fluorescein isothiocyanate), respectively after thawing semen at 50℃ for 12 seconds. The results were analyzed by ANOVA with STATVIEW statistical program. The vitrificatioin solution (LEY + 10.5% glycerol + 1.5% OEP) presented higher motility (20.9%) than other solutions while the solution (LEY + 1M trehalose + 1.5% OEP) showed the lowest (motility : 5.2%). However, survival rates of vitrified sperms detected by FITC showed 1~4% live sperms in almost of dead sperms at all vitrification solutions’ groups, but survival rate of freezing solution of LEY + 1M trehalose + 1.5% OEP LEY and LEY + 10.5% glycerol + 1.5% OEP were showed 49%, and 79%, respectively. There were differences (P<0.05) survival rate of conventional freezing in LEY + 10.5% glycerol + 1.5% OEP and LEY + 1M trehalose + 1.5% OEP and the remaining showed no differences. The results suggested that vitrified boar semen was not enough to be utilized for the artificial insemination, but it showed possibility to utilize for ICSI and conventional freezing with glycerol would be useful method for artificial insemination in pig while we choose the outstanding semen against tolerance to freezing damages.