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Effects of Triladyl-egg Yolk Diluents on the Viability of Frozen Korean Black-goat Spermatozoa from Cauda Epididymis and Electro-ejaculated Semen

Triladyl-난황 희석제가 한국 재래 흑염소의 정소상체 및 전기자극 유래 정자의 융해 후 생존성에 미치는 영향

  • Kim, Sung Woo (Animal Genetic Resources Research Center, National Institute of Animal Science, RDA) ;
  • Lee, Jinwook (Animal Genetic Resources Research Center, National Institute of Animal Science, RDA) ;
  • Kim, Kwan-woo (Animal Genetic Resources Research Center, National Institute of Animal Science, RDA) ;
  • Kim, Chan-Lan (Animal Genetic Resources Research Center, National Institute of Animal Science, RDA) ;
  • Jeon, Ik Soo (Animal Genetic Resources Research Center, National Institute of Animal Science, RDA) ;
  • Lee, Sung-soo (Animal Genetic Resources Research Center, National Institute of Animal Science, RDA)
  • 김성우 (농촌진흥청 국립축산과학원 가축유전자원센터) ;
  • 이진욱 (농촌진흥청 국립축산과학원 가축유전자원센터) ;
  • 김관우 (농촌진흥청 국립축산과학원 가축유전자원센터) ;
  • 김찬란 (농촌진흥청 국립축산과학원 가축유전자원센터) ;
  • 전익수 (농촌진흥청 국립축산과학원 가축유전자원센터) ;
  • 이성수 (농촌진흥청 국립축산과학원 가축유전자원센터)
  • Received : 2017.08.13
  • Accepted : 2017.09.15
  • Published : 2017.09.30

Abstract

To preserve genetic materials, cryopreservation of the semen from live animals is the main technique to establish cryo-banking system which could be used for artificial insemination and embryo transfer. However, the population of Korean black goat (KBG) becomes to dwindle in number and is now faced genetic erosion by crossbreeding with non-native breeds in small KBG farms. In this study, simple freezing method was used to preserve frozen semen from KBG using spermatozoa of cauda epididymis (CE) and electro-stimulated semen (ES). The negative effects of seminal plasma on fresh sperm was confirmed using precipitation test of Triladyl egg yolk diluent and sperm viability after thawing was compared between CE and ES spermatozoa. When seminal plasma of fresh ES semen was washed with semen washing media (SWM), the rates of live sperm shown no significant difference between CE and ES spermatozoa before freezing. However, the survival rate of frozen/thawed CE sperm was higher than ES ($74.6{\pm}10.6%$ vs $53.8{\pm}5.2%$) with significant difference (p < 0.05). The results of longevity test on frozen/thawed sperm from CE showed healthier sperm than ES. Therefore, spermatozoa from CE could be used for cryo-banking system in KBG lines. The more studies are needed to increase survival rate of ES semen.

가축유전자원으로서 흑염소 정액은 가축의 증식 및 유전적 개량을 위하여 동결정액으로서 보존될 필요성이 높으나 아직까지 연구 결과가 많이 누적되어 있지 않은 것으로 판단된다. 국내의 흑염소 동결유전자원의 생산성과 효율성 분석을 위하여 가축유전자원센터에서 보유 중인 흑염소 3 계통을 이용하여 전기자극방법에 의하여 채취된 정자와 정소상체유래 정자를 동결 보존하였다. 동결 전 Triladyl 난황희석액을 이용하여 흑염소의 정액과 혼합하여 $17^{\circ}C$에서 2시간 보존하면 응고현상을 관찰할 수 있었으며 42.9%의 비율로 난황이 변화한다는 것을 육안으로 관찰할 수 있었다. 정장 제거용 배양액으로 전기자극 유래 정자를 세정 후 동결 보존하면 융해 후 정자 생존율이 정액 세정용 배양액 $53.8{\pm}5.2%$로 나타났으며 정소상체 정자는 $74.6{\pm}10.6%$로 유의적으로 높게 관찰되었다. 융해된 정자의 장수성을 $17^{\circ}C$에서 조사해 보면 정소상체 유래 정자가 전기자극 유래 정자보다 우수한 것으로 조사되었다. 그러므로, 염소 동결 유전자원을 보존하는 방법으로 정소상체 정자는 활용성이 인정되며, 염소의 개량과 선발을 위하여 농가에서 활용 가능한 동결정액의 생산과 융해 후 생존성 증진을 위하여 더 많은 동결 보존 연구가 필요하다고 판단된다.

Keywords

References

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