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Effects of concentration and permeation time of cryoprotectants on fertilization and hatching rate in the unfertilized egg of the Pacific oyster Crassostrea gigas
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  • Journal title : The Korean Journal of Malacology
  • Volume 31, Issue 3,  2015, pp.179-186
  • Publisher : The Malacological Society of Korea
  • DOI : 10.9710/kjm.2015.31.3.179
 Title & Authors
Effects of concentration and permeation time of cryoprotectants on fertilization and hatching rate in the unfertilized egg of the Pacific oyster Crassostrea gigas
Kim, Ki Tae; Lim, Han Kyu;
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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.
Cryoprotectant;Fertilization rate;Permeation;Crassostrea gigas;
 Cited by
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. crossref(new window)

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.

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.

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)

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. crossref(new window)

Haga, Y. (1982) On the subzero temperature preservation of fertilized eggs of rainbow trout. Bull. Japan Soc. Sci Fish., 48: 1569-1572. crossref(new window)

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. crossref(new window)

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]

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] crossref(new window)

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.

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,

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]

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]

Renard, P. (1991) Cooling and freezing tolerances in embryo of the Pacific oyster, Crassostrea gigas: methanol and sucrose effects. Aquaculture, 92: 43-57. crossref(new window)

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.

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. crossref(new window)

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. crossref(new window)

Shelton, J.N. (1992) Factors affecting viability of fresh and frozen-thawed sheep embryos. Theriogenology, 37: 713-721. crossref(new window)

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. crossref(new window)

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.

Tervit, H.R. and Gold, P.G. (1984) Deep-freezing of sheep embryos. Theriogenology, 21: 268. crossref(new window)

한국수정란이식 학회 (1995) 소 수정란 이식. pp. 13-132, 정문각. 서울.