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Development of Semen Transport System for Cryopreservation and Fertility in Bull Sperm

소 정자의 동결 및 수정능력 향상을 위한 정액운반법의 개발

  • Lee, Sang-Hee (College of Animal Life Sciences, Kangwon National University) ;
  • Song, Eun-Ji (College of Animal Life Sciences, Kangwon National University) ;
  • Woo, Jea-Seok (National Institute of Animal Science, Hanwoo Ex.) ;
  • Lee, Seung-Hwan (National Institute of Animal Science, Hanwoo Ex.) ;
  • Kang, Hee-Seol (National Institute of Animal Science, Hanwoo Ex.) ;
  • Cheong, Hee-Tae (School of Veterinary Medicine, Kangwon National University) ;
  • Yang, Boo-Keun (College of Animal Life Sciences, Kangwon National University) ;
  • Park, Choon-Keun (College of Animal Life Sciences, Kangwon National University)
  • 이상희 (강원대학교 동물생명과학대학) ;
  • 송은지 (강원대학교 동물생명과학대학) ;
  • 우제석 (국립축산과학원 한우시험장) ;
  • 이승환 (국립축산과학원 한우시험장) ;
  • 강희설 (국립축산과학원 한우시험장) ;
  • 정희태 (강원대학교 수의학과) ;
  • 양부근 (강원대학교 동물생명과학대학) ;
  • 박춘근 (강원대학교 동물생명과학대학)
  • Received : 2013.08.28
  • Accepted : 2013.09.06
  • Published : 2013.09.30

Abstract

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.

Keywords

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