The purpose of this study was to improve the frozen-thawed sperm of Korean Native Cattle using magnetized water. The semen was collected by artificial vagina. Before cryopreservation, Triladyl was flowed through magnet [0, 2000, 4000 and 6000 Gauss (G)] for 5min. The semen was diluted to magnetized Triladyl with 20% egg-yolk for freezing. The diluted semen was placed in 0.5ml straws, and freezing process was exposed on ‒120℃ for 10min. Diluted sperm was stored into LN2. Analysis of frozen- thawed sperm was estimated of viability with SYBR14/PI stain, and membrane intact with hypoosmotic swelling test (HOST). The mitochondrial function analysis was conducted by staining with Rhodamin 123 by flow cytometry and was analyzed histogram. The intensity of acrosome damage was analyzed using FITC-PNA stain by flow cytometry. Analysis of rhodamin 123 and FITC-PNA stain were used mitochondria and acrosome membrane intact. As a results, sperm viability was significantly higher in 4000G (76.0±1.2%) group than other groups (p<0.05). However, HOST was significantly higher in 4000 (37.7±0.6%) and 6000G (35.0±1.1%) than 0 (30.3±0.9%) and 2000G (30.7±0.5%) (p<0.05). In addition, mitochondria membrane and acrosome damage were significantly lower in 6000G (1.40±0.08% and 26.7±2.9%) group than other groups (p<0.05). In conclusion we suggest that magnetized water could improve the ability of sperm on cryopreservation of korean native cattle. * This work was supported by Grant No. PJ 907008 from Rural development administration (RDA).

The purpose of this study was to improve of frozen-thawed sperm using magnetized water in Korean native cattle. Before cryopreservation, without egg-yolk Triladyl® solution was flowed though magnet [0, 2000, 4000 and 6000 Gauss(G)] for 5 min. The freezing of dilluted semen added with Triladyl containing 20% egg-yolk. Analysis of frozen-thawed sperm was estimated viability with SYBR14/PI double stain, membrane intact with hypoosmotic swelling test (HOST), acrosome reaction with FITC-PNA, mitochondria membrane function with Rhodamin 123 by flow- cytometry. Sperm viability was significantly higher in 4000G group than other groups (p<0.05). However, the Hypoosmotic Swelling Test(HOST) was significantly higher in fresh, 4000 and 6000G than 0 and 2000G (p<0.05). In addition, mitochondria membrane damage and acrosome damage were significantly lower in 6000G group than other groups (p<0.05). in conclusion we suggest that magnetized water could be improve ability of sperm on cryopreservation in Korean native cattle.

The objective of this study was to evaluate efficiency of methyl-beta-cyclodextrin (MBCD) in the sperm preservation of bull. For this study, the freezing of diluted semen were added with Triladyl containing 20% egg-yolk and/or 0, 1, 5, 10 and 20 mM MBCD before freezing process. Analysis of viability in frozen-thawed sperm was estimated by SYBR14/PI double stain, hypoosmotic swelling test(HOST) and acrosome damage with FITC-PNA, and mitochondria activation with Rhodamin123 by flow-cytometry. The sperm viability was significantly higher in 0 mM and 5 mM concentrations of MBCD than other groups (p<0.05). However, the HOST was significantly lower at 20 mM concentration of MBCD than other concentrations (p<0.05). In addition, acrosome damage and mitochondria activation rates were significantly lower at 20 mM concentration of MBCD than other groups (p<0.05). In conclusion, the viability of sperm was not significantly different among concentrations of MBCD 0, 5 and 10 mM, but MBCD 20 mM was significantly lower than other groups. In addition, as concentrations of MBCD was high, HOST, acrosome damage and mitochondria activation rates had a negative effect in bull sperm.