한국패류학회지 (The Korean Journal of Malacology)

한국패류학회 (The Malacological Society of Korea)
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동해방지제의 종류, 농도 및 침투시간이 굴 (Crassostrea gigas) 미수정란의 수정률과 부화율에 미치는 영향

Effects of concentration and permeation time of cryoprotectants on fertilization and hatching rate in the unfertilized egg of the Pacific oyster Crassostrea gigas

  • 김기태 (국립수산과학원 남동해수산연구소) ;
  • 임한규 (국립목포대학교 해양수산자원학과)
  • 투고 : 2015.09.11
  • 심사 : 2015.09.30
  • 발행 : 2015.09.30
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초록

굴 미수정란의 동결보존에 적합한 동해방지제의 종류, 농도 및 침투 시간을 구명하기 위하여 4가지 동해방지제 (DMSO, EG, methanol, PEG) 를 대상으로 수정률, 부화율 및 형태 이상률을 조사하였다. 동해방지제의 농도 및 침지시간은 각각 5, 10, 15, 20%와 10, 20, 30, 40분이었다. 침지 후 미수정란은 정상 정자와 수정되었고, 수정률, 부화율 및 형태 이상률은 동해방지제의 종류와 농도 및 침지시간에 영향을 받았다. 동해방지제의 농도와 침지시간의 증가에 따라 수정률과 부화율은 낮아졌고, 대부분의 동해방지제에서 5-10% 농도와 10-20분의 짧은 침지시간에서 좋은 결과를 보였다. 수정률과 부화율을 근거로 굴 미수정란에 가장 적합한 동해방지제는 DMSO였으며, 처리 농도와 침지 시간은 각각 5%와 10-20분이었다.

The Pacific oyster Crassostrea gigas oocytes were exposed to 4 cryoprotectants, dimethyl sulfoxide (DMSO), ethylene glycol (EG), methanol, or polyethylene glycol (PEG), each with 4 four concentrations (5, 10, 15, and 20%) and for 10, 20, 30 or 40 minutes for permeation. The oocytes were then fertilized, using normal sperm of the species. Fertilization and hatching rates were clearly influenced by cryoprotectant species and their concentration and permeation time. Overall, they decreased as concentrations and permeation time of cryoprotectants increased with optimum results at concentrations of 5-10% and a permeation time of 10 minutes. Larval abnormalities, a sign of the chemical damage, were a representative phenotype which was higher at a higher concentration and longer duration of the chemicals. In conclusion, best result was from 5% DMSO exposure for 10-20 minute permeation.

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참고문헌

  1. Chao, N.H., Chiang, C.P., Hsu, H.W., Tsai, C.T. and Lin, T.T. (1994) Toxicity tolerance of oyster embryos to selected cryoprotectants. Aquat Living Res, 7: 99-104. https://doi.org/10.1051/alr:1994013
  2. Chao, N.H., Hsu, H.W., Chang, H.W. and Chen, Y.J. (1996) Finger fish (Monodactylus sebae) as model fish in embryo cryopreservation. COA Fish. Ser. (Council of Agriculture, Taipei), 56: 159-171.
  3. Choi, Y.H. and Chang, Y.J. (1999) Survival rates of trochophores from pearl oyster, Pinctada fucata martemsII and pacific oyster, Crassostrea gigas immersed in four kinds of cryoprotectant. J. Kor. Fish. Soc., 32: 476-480.
  4. Chung, J.K., Lim, H.K, Son, M.H, Kim, J.H. and Jeong, M.H. (2011) Effects of concentration and Immersion time of three cryoprotective agents on the embryos development of abalone Haliotis discus hannai. Development and Reproduction, 15: 337-343. (in Korean)
  5. Cseh, S., Corselli, j., Nehlsin-Cannarella, S.L., Bailey, L.L. and Szalay, A.A. (1997) The effect of quick-freezing in ethylene glycol on morphological survival and in vitro development of mouse embryos frozen at different preimplantation stages. Theriogenology, 48: 43-50. https://doi.org/10.1016/S0093-691X(97)00190-8
  6. Haga, Y. (1982) On the subzero temperature preservation of fertilized eggs of rainbow trout. Bull. Japan Soc. Sci Fish., 48: 1569-1572. https://doi.org/10.2331/suisan.48.1569
  7. Harvey, B.R., Kelley, N. and Ashwood-Smith, M.J. (1983) Permeability of intact and dechroionated zebra fish embryos to glycerol and dimethyl sulfoxide. Cryobiology, 20: 432-439. https://doi.org/10.1016/0011-2240(83)90033-0
  8. Kim, K. T. (2015) Reproductive physiology of pacific oyster Crassostrea gigas and cryopreservation of oysters and larvae. Ph.D. thesis, Pukyong National University, Pusan, Korea, 155 pp. [in Korean with English abstract]
  9. Kim, K.T., Lim, H.K. and Chang, Y.J. (2013) Survival rates with time course of frozen-thawed Pacific oyster larvae in indoor rearing system. Development and Reproduction, 17: 337-343. [in Korean with English abstract] https://doi.org/10.12717/DR.2013.17.4.337
  10. Leibo, S.P., Mcgrath, J.J. and Cravalho, E.G. (1978) Microscopic observation of intracellular ice formation in unfertilized mouse ova as a function of cooling rate. Cryobiology, 15: 157-271.
  11. Leung, L.K.P. and Jamieson, B.G.M. (1991) Live preservation of fish gametes. In: Fish Evolution and Systematics: Evidence from Spermatozoa. (ed. by B.G.M. Jamieson), pp. 245-269. Cambridge University Press, New York,
  12. Lim, H. K. (1998) Physiological properties of sperm and gamete preservation in black seabream, Acanthopagrus schlegeli. Ph.D. thesis, Pukyong National University, Pusan, Korea, 130 pp. [in Korean with English abstract]
  13. Lim, H.K., Chang Y.J. and Jo P.G. (2003) Effects of cryoprotectants on survival and hatching of black seabream, Acanthopagrus schlegeli embryos. J. of Aquaculture, 16: 262-266. [in Korean with English abstract]
  14. Renard, P. (1991) Cooling and freezing tolerances in embryo of the Pacific oyster, Crassostrea gigas: methanol and sucrose effects. Aquaculture, 92: 43-57. https://doi.org/10.1016/0044-8486(91)90007-T
  15. Renard, P. and Cochard, J.C. (1989) Effect of various cryoprotectants on Pacific oyster Crassostrea gigas Thunberg, Manila clam, Ruditapes philippinarum Reeve and King scallop, Pecten maximus (L.) embryo: influence of the biochemical and osmotic effects. Cryo Letters, 10: 169-180.
  16. Sakul, H., Bradford, G.E., Bondurant, R.H., Anderson, G.B. and Donahue, S.E. (1993) Cryopreservation of embryos as means of germ plasm conservation in sheep. Theriogenology, 39: 401-409. https://doi.org/10.1016/0093-691X(93)90383-G
  17. Schiewe, M.C., Rall, W.F., Stuart, L.D. and Wildt, D.E. (1991) Analysis of cryoprotectant, cooling rate and in situ dilution using conventional freezing or vitrification for cryopreserving sheep embryos. Theriogenology, 36: 279-293. https://doi.org/10.1016/0093-691X(91)90386-R
  18. Shelton, J.N. (1992) Factors affecting viability of fresh and frozen-thawed sheep embryos. Theriogenology, 37: 713-721. https://doi.org/10.1016/0093-691X(92)90150-P
  19. Somgsasen, N., Buckrell, B.C., Plante, C. and Leibo, S.P. (1995) In vitro and in vivo survival of cryopreserved sheep embryos. Cryobiology, 32: 78-91. https://doi.org/10.1006/cryo.1995.1007
  20. Suzuki, T., Komada, H., Takai, R., ArII, K. and Kozima, T.T. (1995) Relation between toxicity of cryoprotectant DMSO and its concentration in several fish embryos. Fisheries science, 6: 193-197.
  21. Tervit, H.R. and Gold, P.G. (1984) Deep-freezing of sheep embryos. Theriogenology, 21: 268. https://doi.org/10.1016/0093-691X(84)90368-6
  22. 한국수정란이식 학회 (1995) 소 수정란 이식. pp. 13-132, 정문각. 서울.

피인용 문헌

  1. Effects of cryoprotective agents and treatment methods on sperm cryopreservation of stone flounder, Kareius bicoloratus vol.531, pp.None, 2015, https://doi.org/10.1016/j.aquaculture.2020.735969