Analysis of Embryo Recovery Rate by Direct Embryo Collection (DEC) in Korean Native Cattle (Hanwoo)

한우에서 Direct Embryo Collection(DEC)을 이용한 체내 수정란의 회수율 분석

  • Yoo, Han-Jun (Developmental Biotechnology Laboratory, Myung-poomHanwooConsulting) ;
  • Park, Joung-Jun (Developmental Biotechnology Laboratory, Myung-poomHanwooConsulting) ;
  • Yoon, Pil-Sang (Developmental Biotechnology Laboratory, Myung-poomHanwooConsulting) ;
  • Kim, Ki-Won (Developmental Biotechnology Laboratory, Myung-poomHanwooConsulting) ;
  • Park, Choon-Keun (College of Animal Life Sciences, Kangwon National University)
  • Received : 2011.08.06
  • Accepted : 2011.08.22
  • Published : 2011.09.30

Abstract

This study was performed in order to determine optimum flushing solution using the direct embryo collection (DEC). Donors, at random stages of the estrous cycle, received a CIDR. 7 days later, 200 mg FSH was treated with 40, 30, 20, 10 mg FSH levels in declining doses twice daily by intramuscular injection for 4 days. On the 3$^{rd}$ day administration of FSH, 25 mg $PGF_2{\alpha}$ was administered and CIDR was withdrawn. After FSH injections were complete, donors were artificially inseminated twice at 12 hr intervals. The donor cattle received 250 ${\mu}g$ GnRH at time of 1$^{st}$ insemination and embryos were recovered 8 days after the 1$^{st}$ insemination. Embryo collection from superovulated donors were performed to flushing by DEC and conventional method. As a results, the average number of recovered embryos were significantly higher as 19.1${\pm}$1.40 with DEC method than 12.0${\pm}$0.44 with conventional embryo collection method, respectively (p<0.05). Also, The average number of transferable embryos were significantly higher (p<0.05) as 15.8${\pm}$1.72 with DEC method than 6.9${\pm}$0.35 from conventional embryo recovery procedures. Meanwhile, number of recovered embryos and number of recovered transferable embryos following the number of flushing times until 6${dr}$ flushing were significantly higher as 8.6${\pm}$0.53 and 8.6${\pm}$0.53 from 2$^{nd}$ flushing time than other groups (p<0.05). No. of Ear. B stage embryos were significantly higher as 3.9${\pm}$0.90 and 3.9${\pm}$0.90 with 2$^{nd}$ flushing time in total collected embryos and transferable embryos (p<0.05). Com M stage embryos were significantly higher as 3.7${\pm}$1.00 in 2$^{nd}$ flushing time and as 2.2${\pm}$0.76 in 3$^{rd}$ flushing time for recovered embryos (p<0.05). In transferable embryos, Com. M stage embryos were significantly higher (p<0.05) as 3.7${\pm}$1.00 in 2$^{nd}$ flushing time and as 2.2${\pm}$0.76 in 34$^{dr}$ flushing time, also. No. of degradation embryos was significantly higher as 2.2${\pm}$0.72 in 5${rd}$ flushing time, On the other hand, degradation embryos was not observed in transferable embryos (p<0.05). In conclusion, these results suggest that DEC method should effective methods for production of in vivo embryos using less flushing solution following perform until 4$^{rd}$ flushing time than conventional embryo collecting method. Also, it might be effectively collection of transferable embryos following more less procedure times compared to conventional embryo recovery methods.

Keywords

References

  1. 김덕임, 서상원, 정재경, 이규승, 서길웅, 박창식, 정영채, 박병권. 2002. 한우에 있어서 체내 수정란의 생산과 이식에 관한 연구: 한우 수정란 이식이 수태율에 미치는 요인. 한국수정란이식학회지 17:33-44. https://doi.org/10.1016/j.anireprosci.2004.04.034
  2. 박성재, 양보석, 임기순, 성환후, 장원경, 정일정, 박충생. 2000.한우에 있어서 초음파기기를 이용한 생체내 난자 채취시 채란조건 및 수태율에 관한 연구. 한국수정란이식학회지 24:199-208. https://doi.org/10.1016/S0093-691X(97)00303-8
  3. 박용수, 김소섭, 박흠대, 박현정, 김재명. 2005. 한우 체외 수정란이 이식된 수란우의 임신과 유산에 영향을 미치는 수정란측 요인. 한국수정란이식학회지 20:89-95. https://doi.org/10.1016/0093-691X(91)90153-5
  4. 손동수, 김일화, 류일선, 연성흠, 서국현, 이동원, 최선호, 박수봉, 이충섭, 최유림, 안병석, 김준식. 2000. 젖소MOET Scheme의 추진을 위한 수정란 생산과 이식. 한국수정란이식학회지 15:57-65.
  5. 임석기, 우제석, 전기준, 장선식, 강수원, 윤상기, 손동수. 1998.한우에 있어서 PEG에 용해시킨 Folltropin-V의 1회 피하주사에 의한 다배란 유기. 한국수정란이식학회지 13:207-212.
  6. 진종인, 권태현, 최병현, 김성수, 조현태, 공일근. 2010. OPU(OvumPick-Up) 채란기간이 난자 및 수정란 생산에 미치는 영향.한국수정란이식학회지 25:15-20. https://doi.org/10.1016/0093-691X(94)00013-K
  7. 최병헌, 진종인, 권태현, 김성수, 조현태, 공일근. 2011. 한우와젖소 대리모가 OPU 유래 한우 송아지의 체중과 임신 기간에 미치는 영향. 한국수정란이식학회지 26:27-32. https://doi.org/10.1016/S0093-691X(99)00236-8
  8. 최선호, 류일선, 손동수, 조상래, 한만희, 김현종, 최창용, 김영근. 2005. 한우의 반복 과배란 및 산차가 수정란 생산에 미치는 영향. 한국수정란이식학회지 20:185-190. https://doi.org/10.1016/0093-691X(94)90384-U
  9. Ax RL, Armbrust S, Tappan R, Gilbert G, Oyarzo JN, Bellin ME, Selner D and McCauley TC 2005. Superovulation and embryo recovery from peripubertal Holstein heifer. Anim. Reprod. Sci. 85:71-80. https://doi.org/10.1095/biolreprod.102.004481
  10. Blondin P, Guibault LA and Sirard MA. 1997. The time interval between FSH-P administration and slaughter can influence the developmental competence of beef heifer oocytes. Theriogenology 48:803-813.
  11. Christensen LG. 1991. Use of embryo transfer in future cattle breeding schemes. Theriogenology 35:141-149. https://doi.org/10.1016/j.theriogenology.2005.09.028
  12. Donaldson L. 1983. The effect of prostagladin F2 alpha traetments in superovulated cattle on estrus response and embryo production. Theriogenology 20:279-285. https://doi.org/10.1016/S0378-4320(02)00241-5
  13. Gilmour AR, Gogel BJ, Cullis BR and Thompson R. 2006. ASReml User Guide Release 2.0 VSN. International Ltd, Hemel Hempstead, HP1 1ES, UK. https://doi.org/10.1016/S0093-691X(02)01246-3
  14. Greve T, Callesen H, Hyttel H, Hoier R and Assey R. 1995. The effects of exogenous gonadotropins on oocyte and embryo quality in cattle. Theriogenology 43:41-50. https://doi.org/10.1016/S0093-691X(00)00463-5
  15. Hendriksen PJ, Vos PL, Steenweg WN, Bevers MM and Dieleman SJ. 2000. Bovine follicular development and its effect on the in vitro competence of oocytes. Theriogenology 53: 11-20. https://doi.org/10.1016/0093-691X(88)90310-X
  16. Kruip TA, Boni R, Wurth YA, Roelofsen MW and Pieterse MC. 1994. Potential use of ovum pick-up for embryo production and breeding in cattle. Theriogenology 42:675-684. https://doi.org/10.1002/mrd.1153
  17. Lazzari G, Wrenzycki C, Herrmann D, Duchi R, Kruiop T, Niemann H and Galli C. 2002. Cellular and molecular deviations in bovine in vitro produced embryos are related to the large offspring syndrome. Biol. Reprod. 67:767-775.
  18. Lohuis MM. 1997. Strategy for dairy cattle improvement utilizing MOET in Canada. Anim. Genetic and Breeding 1: 224-226. https://doi.org/10.1016/0093-691X(96)00174-4
  19. Lonergan P, Fair T, Corcoran D and Evans AC. 2006. Effect of culture environment on gene expression and developmental characteristics in IVF-derived embryos. Theriogenology 65: 137-152. https://doi.org/10.1126/science.7194504
  20. Manik RS, Singla SK and Palta P. 2003. Collection of oocytes through transvaginal ultrasound-guided aspiration of follicles in an Indian breed of cattle. Anim. Reprod. Sci. 76: 155-161. https://doi.org/10.1016/0093-691X(88)90040-4
  21. Merton JS, de Roos AP, Mullaart E, de Ruigh L, Kaal L, Vos PL and Dieleman SJ. 2003. Factors affecting oocyte quality and quantity in commercial application of embryo technologies in the cattle breeding industry. Therogenology 59: 651-674.
  22. Numabe T, Oikawa T, Kikiuchi T and Horiuchi T. 2000. Production efficiency of Japanese Black calves by transfer of produced in vitro. Theriogenology 54:1409-1420. https://doi.org/10.1016/S0093-691X(98)00038-7
  23. Pieterse MC, Kappen KA, Kruip TA and Taverne MA. 1988. Aspiration of bovine oocytes during transvaginal ultrasound scanning of the ovaries. Theriogenology 30:751-762. https://doi.org/10.1016/S0093-691X(99)00259-9
  24. Rizos D, Ward F, Duffy P, Boland MP and Lonergan P. 2002. Consequences of bovine oocyte maturation fertilization or early emrbryo development in vitro versus in vivo: Implications for blastocyst yield and blastocyst quality. Mol. Reprod. Dev. 61:234-248.
  25. Rowe RF, Del Campo MR, Critser JK and Ginther OJ. 1980. Embryo transfer in cattle : Nonsurgical transfer. Am. J. Vet. Res. 41:1024-1028.
  26. Schmidt M, Greve T, Avery B, Beckers JF, Sulon J and Hansen HB. 1996. Pregnancies, calves and calf viability after transfer of in vitro produced bovine embryos. Theriogenology 46:527-539.
  27. Seidel GE. 1981. Superovulation and embryos transfer in cattle. Science 211:351-358.
  28. Smith C. 1988. Application of embryo transfer in animal breeding. Theriogenology 29:203-212.
  29. Stringfello DA and Seidel SM. 1998. Manual of the International Embryo Transfer Society. 3dr ed., International Embryo Transfer Society Inc., Illinois, pp. 165-170.
  30. van Wagtendonk-de Leeuw AM, Aerts BJ and den Dass JH. 1998. Abnormal offspring following in vitro production of bovine preimplantation embryos: A field study. Theriogenology 49:883-894.
  31. van Wagtendonk-de Leeuw AM, Mullaart E, de Roos AP, Merton JS, Den Daas JH, Kemp B and de Ruigh L. 2000. Effects of different reproduction techniques: AI, MOET or IVP, on health and welfare of bovine offspring. Theriogenology 53:575-597.