Korean Journal of Poultry Science (한국가금학회지)

The Korean Society of Poultry Science (한국가금학회)
권호 표지
DOI QR코드

DOI QR Code

Effect of N-Methylacetamide Concentration on the Fertility and Hatchability of Cryopreserved Ogye Rooster Semen

N-Methylacetamide 동결 보호제의 농도가 오계 동결 정액의 수정 및 부화율에 미치는 영향

  • Kim, Sung Woo (Animal Genetic Resauces Station, National Institute of Animal Science, RDA) ;
  • Choi, Jin Seok (Animal Genetic Resauces Station, National Institute of Animal Science, RDA) ;
  • Ko, Yeoung-Gyu (Animal Genetic Resauces Station, National Institute of Animal Science, RDA) ;
  • Do, Yoon-Jung (Animal Genetic Resauces Station, National Institute of Animal Science, RDA) ;
  • Byun, Mijeong (Animal Genetic Resauces Station, National Institute of Animal Science, RDA) ;
  • Park, Soo-Bong (Animal Breeding and Genetics Division, National Institute of Animal Science, RDA) ;
  • Seong, Hwan-Hoo (Animal Genetic Resauces Station, National Institute of Animal Science, RDA) ;
  • Kim, Chong-Dae (Poutry Science Division, National Institute of Animal Science, RDA)
  • 김성우 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 최진석 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 고응규 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 도윤정 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 변미정 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 박수봉 (농촌진흥청 국립축산과학원 개량평가과) ;
  • 성환후 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 김종대 (농촌진흥청 국립축산과학원 가금과)
  • Received : 2013.12.18
  • Accepted : 2013.02.19
  • Published : 2014.03.31
Download PDF
⟨ Previous Next ⟩

Abstract

To preserve chicken genetic materials like cryopreserved spermatozoa, various kinds of freezing agents like glycerol, dimethylsuloxide, dimethylformamide or dimethylacetamide have been used for rooster semen preparation. Recently, the usage of N-methylacetamide (MA) for Ogye rooster semen preservation resulted in hatched chicken successfully. In this study, we investigated the effects of 7, 9 and 11% of MA on the viability, fertility and hatchability of frozen-thawed rooster semen using artificial insemination. The results of viability, fertility and hatchability in frozen semen with 7%, 9% or 11% MA were $35.16{\pm}6.12%$, $67.83{\pm}15.3%$ and $66.2{\pm}16.3%$ of motile sperm rate, 21.5%, 34.7% and 25% of fertility rate, and 100%, 89.5% and 87.5% of hatchability rate. The results of control group with frozen semen were 96.0% of fertility rate and 92.2% of hatchability rate. With these results, the concentration range of MA as a freezing agent of rooster semen could be 7~9% of media. The higher concentration of 9 % MA could decrease the fertility rate of thawed semen not the rate of hatchability rate. So the use of MA without affecting fertility rate would be a key point of freezing method of rooster semen for poultry genetic resource preservation.

본 연구는 닭 정액의 동결 보존을 위하여 비 글리세롤성 동결 보호제 중 MA 농도가 정자의 생존율과 융해된 동결 정자를 인공수정을 실시하여 생산된 수정란의 수정율, 부화율을 조사하고자 실시하였다. 동결 보조제로써 MA의 효율성은 7%, 9% 및 11% 범위에서 동결을 실시하였을 때, 정자의 생존율은 $35.16{\pm}6.12%$, $67.83{\pm}15.3%$ 및 $66.2{\pm}16.3%$로 관찰되었으며, 융해된 정자를 인공수정을 실시하여 생산된 수정란의 수정율은 21.5%, 34.7% 및 25%로 관찰되었으며, 수정된 수정란의 부화율은 100%, 89.5% 및 87.5%로 관찰되었다. 대조군으로써 신선 정액은 수정율이 96.0%로 관찰되었고, 부화율은 92.2%로 관찰되었다. 9% MA를 이용한 간이 동결법으로 생산된 동결 정자를 이용하여 3주간 수정란을 검사하였을 때, 수정율은 비록 35.3%로 관찰되었으나, 부화율은 90.3%로 관찰되었다. 이러한 결과에 따르면, 9% 농도로 MA 동결 보호제를 이용할 경우, 동결 및 융해된 정자를 이용하여 생산된 수정란에서 수정율을 감소시킬 수 있음을 보여주고 있으며, 부화율에는 영향을 주지 않음을 추정할 수 있다. 그러므로 수정율과 부화율에 나쁜 영향을 미치지 않고 가금 유전자원의 보존에 중요한 요인이 될 수 있는 적절한 농도의 MA 동결 보호제 범위는 7~9% 농도로 추정된다.

Keywords

References

  1. Allen TE, Bobr LW 1955 The fertility of fowl spermatozoa in glycerol diluent after intrauterine insemination. Poultry Sci 34:1167-1169. https://doi.org/10.3382/ps.0341167
  2. Blesbois E, Brillard JP 2007 Specific features of in vivo and in vitro sperm storage in birds. Animal 1:1477-1481.
  3. Burrows WH, Quinn JP 1935 A method of obtaining spermatozoa from the domestic fowl. Poultry Sci 14:251-254. https://doi.org/10.3382/ps.0140251
  4. Clark CE, Shaffner CS 1960 The fertilizing capacity of frozen chicken sperm and the influence of related in vitro processes. Poultry Sci 39:1213-1220. https://doi.org/10.3382/ps.0391213
  5. Chalah T, Seigneurin F, Blesbois E, Brillard JP 1999 In vitro comparison of fowl sperm viability in ejaculates frozen by three different techniques and relationship with subsequent fertility in vivo. Cryobiology 39:185-191. https://doi.org/10.1006/cryo.1999.2201
  6. Donoghue AM, Holsberger DR, Evenson DP, Froman DP 1998 Semen donor selection by in vitro sperm mobility increases fertility and semen storage in the turkey hen. J Androl 19:295-301.
  7. Donoghue AM, Wishart GJ 2000 Storage of poultry semen. Animal Reproduction Sci 62:213-232. https://doi.org/10.1016/S0378-4320(00)00160-3
  8. Froman DP, McLean DJ 1996 Objective measurement of sperm motility based upon sperm penetration of Accudenz. Poult Sci 75:776-784. https://doi.org/10.3382/ps.0750776
  9. Gill SPS, Buss EG, Mallis RJ 1996 Cryopreservation of rooster semen in thirteen and sixteen percent glycerol. Poultry Sci 75:254-256. https://doi.org/10.3382/ps.0750254
  10. Hanzawa S, Niinomi T, Miyata T, Tsutsui M, Tajima A 2010 Cryopreservation of chicken semen using N-methylacetamide as cryoprotectant. Jp Poult Sci 47:J27-J32.
  11. Hammerstedt RH, Graham JK 1992 Cryopreservation of poultry sperm: The enigma of glycerol. Cryobiology 29:26-38. https://doi.org/10.1016/0011-2240(92)90004-L
  12. Kirby JD, Tressler CJ, Kirby YK 1998 Evaluation of the duration of sperm fertilizing ability in five lines of commercial broiler breeder and Delaware cross males. Poultry Sci 77:1688-1694. https://doi.org/10.1093/ps/77.11.1688
  13. Lake PE, Ravie O 1984 An exploration of cryoprotective compounds for fowl spermatozoa. Br Poult Sci 25:145-150. https://doi.org/10.1080/13632758408454852
  14. Long JA, Kulkarni G 2004 An effective method for improving the fertility of glycerol-exposed poultry semen. Poultry Sci 83:1594-1601. https://doi.org/10.1093/ps/83.9.1594
  15. McLean DJ, Feltmann AJ, Froman DP 1998 Transfer of sperm into a chemically defined environment by centrifugation through 12% (wt/vol) Accudenz. Poult Sci 77:163-168. https://doi.org/10.1093/ps/77.1.163
  16. Neville WJ, Macpherson JW, Reinhart B 1971 The contraceptive action of glycerol in chickens. Poultry Sci 50:1411-1415. https://doi.org/10.3382/ps.0501411
  17. Purdy PH, Song Y, Silversides FG, Blackburn HD 2009 Evaluation of glycerol removal techniques, cryoprotectants, and insemination methods for cryopreserving rooster sperm with implications for breed and/or line regeneration. Poultry Sci 88:2184-2191. https://doi.org/10.3382/ps.2008-00402
  18. Sasaki K, Tatsumi T, Niinomi T, Imai T, Naito M, Tajima A, Nishi Y 2010 A method for cryopreserving semen from Yakido roosters using N-methylacetamide as a cryoprotectant. Jp Poult Sci 47:297-310. https://doi.org/10.2141/jpsa.009111
  19. Schramm GP 1991 Eignung verschiedener gefrierschutzstoffe zur kryoprotektion von hahnensperma.(German) Monatsh Veterinaermedizin 46:438-440.
  20. Sexton TJ 1973 Effect of various cryoprotectants on the viability and reproductive efficiency of chicken spermatozoa. Poultry Sci 52:1353-1357. https://doi.org/10.3382/ps.0521353
  21. Tereshchenko AV 1985 Study of the toxic effects of some cryoprotectants on cock spermatozoa using fluorescent stains. Nauchno-Tekh Bull Ukr Poult Inst 19:32-36.
  22. Tselutin K, Seigneurin F, Blesbois E 1999 Comparison of cryoprotectants and methods of cryopreservation of fowl spermatozoa. Poultry Sci 78:586-590. https://doi.org/10.1093/ps/78.4.586
  23. Wishart GJ 1985 Quantitation of the fertilizing ability of fresh compared with frozen and thawed fowl spermatozoa. Br Poultry Sci 26:375-380. https://doi.org/10.1080/00071668508416825
  24. 박영식 임경순 이용빈 1987 Dimethylacetamide 첨가와 동결융해방식이 닭 동결정액의 정자생존성과 수정율에 미치는 영향. Korean J Anim Sci 29(9):377-382
  25. 최진석 김성우 신단비 고응규 도윤정 김동훈 공일근 박수봉 2012 N-methylacetamide 동결 보호제가 오계 동결정액의 생존성, 수정 및 부화율에 미치는 영향. 한국가금학회지 39(4): 291-295.
  26. 최진석 신단비 고응규 도윤정 변미정 박수봉 성환후 김현 공일근 김성우 2013 닭 정액 동결 시 동결 보호제가 정액 성상에 미치는 영향. 한국가금학회지 40(3): 171-178. https://doi.org/10.5536/KJPS.2013.40.3.171