The objective of this study was to develop of semen transport system for cryopreservation and fertility in bull sperm. The ejaculated semen were diluted with Triladyl containing 20% egg-yolk for transportation. Diluted semen was transported by three methods that there were wrapping tissue (Tissue), sinking under $30^{circ}C$ water (Water) and sinking between warm water and air (Air) methods. Semen was transported within 2 hours in $0.3^{circ}C$. For this study, the freezing of diluted semen were added with Triladyl containing 20% egg-yolk. And frozen-thawed sperm were estimated with SYBR14/PI double stain for viability, FITC-PNA/PI double stain for acrosome reaction analysis and Rhodamine123 double stain for mitochondrial intact assessment. In results, live sperm (SYBR+/PI-) in Air treatment group ($43.3{pm}4.7%$) was significantly (p<0.05) higher than other treatment groups (Tissue: $16.3{pm}2.7%$ and Water: $27.5{pm}3.1%$), dying sperm (SYBR+/PI+) in Air treatment group ($55.6{pm}4.7%$) was significantly lower than other treatment groups (Tissue: $77.6{pm}3.2%$ and Water: $67.6{pm}3.3%$) (p<0.05). Acrosome reaction in Air treatment group ($0.2{pm}0.1%$) within live sperm (PI negative region) was significantly (p<0.05) lower than other treatment groups (Tissue: $0.7{pm}0.2%$ and Water: $0.5{pm}0.1%$), the acrosome reaction in Air treatment group ($28.6{pm}2.8%$) within all sperm also was significantly lower than other treatment groups (Tissue: $44.2{pm}1.8%$ and Water: $36.2{pm}2.0%$) (p<0.05). And mitochondrial intact in Air treatment group within live ($97.1{pm}0.4%$) and all ($61.9{pm}3.3%$) sperm were significantly higher than other treatment groups (Tissue: $85.2{pm}3.3%$, Water: $87.8{pm}2.9%$ within live sperm and Tissue: $49.28{pm}3.7%$, Water: $42.0{pm}3.1%$ within all sperm) (p<0.05). Therefore, we suggest that transportation by sinking method between warm water and air was beneficial to improvement of fertility in frozen-thawed in bull semen.