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ISSN : 1225-3480
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.
Chao, N.H., Chiang, C.P., Hsu, H.W., Tsai, C.T., Lin, T.T. (1994) Toxicity tolerance of oyster embryos to selected cryoprotectants. Aquat Living Res, 7:99-104.
Chao, N.H., Hsu, H.W., Chang, H.W., Chen, Y.J. (1996) Finger fish (Monodactylus sebae) as model fish in embryo cryopreservation. COA Fish. Ser. (Council of Agriculture, Taipei), 56: 159-171.
Choi, Y.H., Chang, Y.J. (1999) Survival rates of trochophores from pearl oyster, Pinctada fucata martemsii, pacific oyster, Crassostrea gigas immersed in four kinds of cryoprotectant. J. Kor. Fish. Soc., 32: 476-480.
Chung, J.K., Lim, H.K, Son, M.H, Kim, J.H., Jeong, M.H. (2011) Effects of concentration, Immersion time of three cryoprotective agents on the embryos development of abalone Haliotis discus hannai. Development, Reproduction, 15: 337-343. (in Korean)
Cseh, S., Corselli, j., Nehlsin-Cannarella, S.L., Bailey, L.L., Szalay, A.A. (1997) The effect of quick-freezing in ethylene glycol on morphological survival, in vitro development of mouse embryos frozen at different preimplantation stages. Theriogenology, 48: 43-50.
Haga, Y. (1982) On the subzero temperature preservation of fertilized eggs of rainbow trout. Bull. Japan Soc. Sci Fish., 48: 1569-1572.
Harvey, B.R., Kelley, N., Ashwood-Smith, M.J. (1983)Permeability of intact, dechroionated zebra fish embryos to glycerol, dimethyl sulfoxide. Cryobiology, 20: 432-439.
Kim, K. T. (2015) Reproductive physiology of pacific oyster Crassostrea gigas, cryopreservation of oysters, larvae. Ph.D. thesis, Pukyong National University, Pusan, Korea, 155 pp. [in Korean with English abstract]
Kim, K.T., Lim, H.K., Chang, Y.J. (2013) Survival rates with time course of frozen-thawed Pacific oyster larvae in indoor rearing system. Development, Reproduction, 17: 337-343. [in Korean with English abstract]
Leibo, S.P., Mcgrath, J.J., Cravalho, E.G. (1978)Microscopic observation of intracellular ice formation in unfertilized mouse ova as a function of cooling rate. Cryobiology, 15: 157-271.
Leung, L.K.P., Jamieson, B.G.M. (1991) Live preservation of fish gametes. In: Fish Evolution, Systematics: Evidence from Spermatozoa. (ed. by B.G.M. Jamieson), pp. 245-269. Cambridge University Press, New York,
Lim, H. K. (1998) Physiological properties of sperm, gamete preservation in black seabream, Acanthopagrus schlegeli. Ph.D. thesis, Pukyong National University, Pusan, Korea, 130 pp. [in Korean with English abstract]
Lim, H.K., Chang Y.J., Jo P.G. (2003) Effects of cryoprotectants on survival, hatching of black seabream, Acanthopagrus schlegeli embryos. J. of Aquaculture, 16: 262-266. [in Korean with English abstract]
Renard, P. (1991) Cooling, freezing tolerances in embryo of the Pacific oyster, Crassostrea gigas:methanol, sucrose effects. Aquaculture, 92: 43-57.
Renard, P., Cochard, J.C. (1989) Effect of various cryoprotectants on Pacific oyster Crassostrea gigas Thunberg, Manila clam, Ruditapes philippinarum Reeve, King scallop, Pecten maximus (L.) embryo:influence of the biochemical, osmotic effects. Cryo Letters, 10: 169-180.