Cryopreservation of fish gametes is an emerging technology and breeding with cryopreserved gametes is advancement in fish seed production. Success of cryopreservation is evaluated by the post - thaw motility of the spermatozoa, an for which energy is required. Cryopreservation is known to cause changes in the seminal plasma that would alter the energy supply for the motility of the spermatozoa. Therefore, energy supplementation is found to be useful during cryopreservation. Cirrhinus mrigala spermatozoa were cryopreserved along with glucose as a co-cryoprotectant after 1:100 dilutions with 0.85% physiological saline as extender and Dimethyl Sulfoxide (DMSO) as cryoprotectant (85:15). The diluents contained glucose at four different concentrations, viz., T1 (0.25%), T2 (0.5%), T3 (0.75%) and T4 (1%). The diluted milt was equilibrated for 10 min at 5Ëš C and loaded into 0.25 ml straws. The loaded straws were then frozen with LN2 vapour for 5 min and immersed in liquid nitrogen. Observations were made once in 7 days for 42 days on motility parameters based on which the duration, score, pattern and percentage were determined. The spermatozoa cryopreserved with glucose at 0.5% concentration showed the highest motility duration of 204Â±3.6 s whereas Control group showed motility duration of only 83.33Â± 4.5 s on 42nd day. The difference in motility duration was statistically significant (P>0.025).The present study revealed the benefits of adding glucose a t0.5% during cryopreservation as it could help in maintaining the motility duration and survival of spermatozoa.
Cirrhinus mrigala, Cryopreservation, Energy supplements, Glucose, Spermatological parameters
Babiak, I., Glogowski, J., Luezynski, M.J., Kucharezyk, D. and Luezynski, M. (1995). Cryopreservation of the milt of the northern pike. J. Fish Biol., 46 (5): 819-828.
Betsy, C.J. (2013). Role of supplemented energy sources on spermatological parameters of selected cultivable carps. M.F. Sc. Thesis, pp 101.
Billard, R., Cosson, J., Perchec, G. and Linhart, O. (1995). Biology of sperm and artificial reproduction in carp. Aquaculture, 124: 95â€“112.
Blaxter, P.C. (1953). Sperm storage and cross fertilization of spring and autumn spawning herring. Nature, 172: 1189-1190.
Bozkurt, Y., Secer, S., Tekin, N., Akcay, E. (2005). Cryopreservation of Rainbow Trout (Oncorhynchus mykiss) and Mirror Carp (Cyprinus carpio) Sperm with Glucose Based Extender. Cilt I Sayn I, 21-25.
Carolsfeld, J., Godinho, H.P., Filho, E.Z. and Harvey, B.J. (2003). Cryopreservation of sperm in Brazilian migratory fish conservation. J. Fish Biol., 63: 472â€“489.
Chao, N.H. and Liao, I.C. (2001).Cryopreservation on finfish and shellfish gametes and embryos. Aquaculture, 197: 161-189.
Chao, N.H., Chan, W.C., Liu, K.C. and Liao, I.C. (1987). The properties of Tilapia sperm and its cryopreservation. J. Fish Biol., 30: 107-119.
Christen, F., Gatti, J.L. and Billard, R. (1987). Trout sperm motility- the transient movement of trout sperm is related to changes in the concentration of ATP following the activation of flagellar movement. Eur. J. Biochem., 166: 667â€“ 671.
Chutia, I.P., Krishna, G., and Chaudhary, A. (1998). Biochemical and biometrical analysis of carp milt. Fish genetics and Biodiversity conservation, Nature conservators, Muzaffarnager, pp 205-213.
Cruz-Casallas, P.E., Pardo-Carrasco, S.C., Arias-Castellanos, J.A., Lombo-Castellanos, P.E., Lombo-RodrÃguez, D.A. and Pardo-MariÃ±o, J.E. (2004). Cryopreservation of yamÃº Brycon siebenthalae milt. J World Aquacult Soc., 35:529-35.
Doi, M., Hoshino, T., Taki, Y. and Ogasawara, Y. (1982). Activity of the sperm of the Bluefin tuna Thunnus thynnus under fresh and preserved conditions. Bull. Japan Soc. Sci. Fish., 48(4): 495-498.
Dong, Q., Huang, C. and Tiersch, T.R. (2006). Post-thaw amendment of cryopreserved sperm for use in artificial insemination of a viviparous fish, the green swordtail Xiphophorus helleri. Aquaculture, 259: 403â€“414.
FAO (2014). Fishery and Aquaculture Statistics. Food and Agriculture Organization of the United Nations, Rome, 2014.
Fribourgh, B.H. (1966). The application of differential staining method to low temperature studies on goldfish spermatozoa. Prog. Fish Cult., 28: 227-231.
Gardiner, D.M. (1978). Utilization of extracellular glucose by spermatozoa of two viviparous fishes. Comparative Biochemistry and Physiology, 59: 165-168.
Glogowski, J., Ciereszko, R. and Dabrowski, K. (1999). Cryopreservation of Muskellunge and Yellow Perch semen. North American J. Aquaculture, 61: 258-262.
Grace, L.B. (2013). Deep freezing trials of Arius arius spermatozoa. J. Appl. Environ. Biol. Sci., 3(6): 42-47.
Guest, W.C., Avault, Jr.J.W. and Roussed, J.D. (1976). Preservation of channel catfish sperm. Trans. Am. Fish. Soc., 105: 469-474.
Hammerstedt, R.H., Graham, J.K. and Nolan, J.P. (1990). Cryopreservation of mammalian sperm: What we ask them to survive. Andrology, 11(3): 73-88.
Hamner, E and Charles, O.A. (1969). Physiological considerations in capacitatum of spermatozoa. In endocrinological de la reproduction. Mexico, D.E. La Prensa Medica Mexican, pp. 24-46.
Harvey, B. and Kelley, R.N. (1984). Chilled storage of Sarotheroden mossambicus milt. Aquaculture, 36: 85-95.
Henderson, N.E. and Dewar, J.E. (1959). Short term storage of brook trout milt. Prog. Fish. Cult., 21: 169-171.
HorvÃ¡th, A., Miskolczi, E., UrbÃ¡nyi, B. (2003). Cryopreservation of common carp sperm. Aquat Living Resour., 16:457-460.
Kruger, J.C.D.W., Smit, G.L., Vuren, J.H.J.V. and Ferreiera, J.T. (1984). Some chemical and physical properties of the semen of Cyprinus carpio L. and Oreochromis mossambicus (Peters). J. Fish Biol., 24: 263-272.
Kurokura, H., Hirano, R., Tomita, M. and Iwahashi, M. (1984). Cryopreservation of carp sperm. Aquaculture, 37: 267-273.
Lahnsteiner, F. and Patzner, R.A. (1998). Sperm motility of the marine teleosts Boops boops, Diploduss argus, Mullus barbatus and Trachurus mediterraneus. J. Fish Biol., 52(4):726-742.
Le Roux, P. and Steyn, L.(1968). In Visse van Transvaal. Kaapstad: Kaapen Transval Drukkers Beperk, pp. 29-30
Leung, L.K.P. and Jamieson, B.G.M. (1991). Live preservation of fish gametes. Fish evaluation and systematic evidence from spermatozoa. University Press, Cambridge, pp 245.
Lubzens, E., Daube, N., Pekarsy, I., Magnus, Y., Cohen, A.,Yusefovich, F. and Feigin, P. (1997). Carp (Cyprinus carpio L) spermatozoa cryobanks- strategies in research and application. Aquaculture, 155: 13-30.
MartÃnez, J.G., Tarazona-Morales, A.M. and Pardo-Carrasco, S.C. (2012). Sperm cryopreservation of freshwater fish bocachico (Prochilodus magdalenae) in DMSO and glucose and its effects on fertilization and hatching efficiency. Anim. Reprod., 9(1): 19-26.
Melo, F.C.S.A. and Godinho, H.P. (2006). A protocol for cryopreservation of spermatozoa of the fish Brycon orthotaenia. Animal Reproduction, 3(3): 380-385.
Mohanraj, P. (2009). Effect of cryopreservation on the irradiated spermatozoa of Cyprinus carpio (Linn.). M.F. Sc. Thesis, pp 71.
Mounib, M.S. (1967). Metabolism of pyruvate, acetate and glyoxylate by fish sperm. Comparative Biochemistry and Physiology, 20: 987-992.
Nascimento, A.F., Maria, A.N., Pessoa, N.O., Carvalho M.A. and Viveiros, A.T. (2010). Out-of-season sperm cryopreserved in different media of the Amazonian freshwater fish pirapitinga (Piaractus brachypomus). Anim. Rep. Sci., 118 (2-4): 324-9.
Nomura, M. (1964). Studies on reproduction of rainbow trout Salmo gairdneri with special reference to egg takens VI- the activities of spermatozoa in different diluents and preservation of senmen. Bull. Japan Soc. Sci. Fish., 30: 723-733.
Ohta, H., Shimma, H. and Hirose, K. (1995). Relationship between fertility and motility of cryopreserved spermatozoa of the amago salmon Oncorhynchus masou ishikawae. Fisheries Science, 61(5): 886-887.
Perchec, G., Jeulin, C., Cosson, J., Andre, F. and Billard, R. (1995). Relationship between sperm ATP content and motility of carp spermatozoa. J. Cell. Sci., 108: 747â€“ 753.
Ponniah, A.G., Lakra, W.S. and Ogale, S.N. (1999). Effects of cryoprotectant and fertilization protocol on viability of Mahseer Tor khudree cryopreserved spermatozoa. J. Aqua Trop., 14(2): 153-158.
Purdy, P.H. (2006). A review on goat sperm cryopreservation. Small Rumin Res, 63:215-225.
Sarder, M.R.I., Rafiquzzaman, S.M., Sultana, R., and Faridul Islam, M. (2009). Cryopreservation of spermatozoa of mrigal, Cirrhinus cirrhosus with a view to minimize inbreeding and hybridization. J. Bangladesh Agri. Univ., 7(1): 211-215.
Stein, H. (1975). Spezielleuntersuchungen am Fischspermaun-terbesonderer Berucksichtigung der spermakonservierung. Dissertation, Tech UniverststMunchen.
Stoss, J. and Refstie, T. (1983). Short-term storage and cryopreservation of milt from Atlantic salmon and Sea Trout. Aquaculture, 30: 229-236.
Takai, H. and Morisawa, M. (1995). Changes in intracellular K+ concentration caused by external osmolality change regulate sperm motility of marine and freshwater teleosts. J. Cell. Sci., 108: 1175â€“1181.
Viveiros, A.T.M., Godinho, H.P. (2009). Sperm quality and cryopreservation of Brazilian freshwater fish species: a review. Fish Physiol Biochem., 35:137-150.
Viveiros, A.T., Nascimento, A.F., OrfÃ£o, L.H. and IsaÃº, Z.A. (2010). Motility and fertility of the subtropical freshwater fish streaked prochilod (Prochilodus lineatus) sperm cryopreserved in powdered coconut water. Theriogenology, 74(4): 551-556.
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