**Part 6**

**Cryopreservation of Aquatic Species** 

388 Current Frontiers in Cryobiology

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Swanson, W. F. (2002). The role of science and reproductive biotechnology in establishing

Swanson, W. F. & Brown, J. L. (2004). International training programs in reproductive

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Tebet, J. M.; Martins, M. I. M; Chirinea, V. H.; Souza, F. F.; Campagnol, D.; Lopes, M. D.

Verhage, H. G.; Beamer, N. B. & Brenner, R. M. (1976). Plasma levels of estradiol and

Wildt, D. E.; Platz, C. C.; Chakraborty, P. K. & Seager, S. W. J. (1979). Oestrus and ovarian

Wildt, D. E.; Platz, C. G.; Seager, S. W. J. & Bush, M. (1981). Induction of ovarian activity in the cheetah *(Acinonyx jubatus)*. *Biology of Reproduction*, Vol.24, pp. 217-222.

electroejaculated spermatozoa. *Theriogenology*, Vol. 66, pp. 1629-1632. Tsuitsui, T. & Stabenfeldt, G. H. (1996). Biology of ovarian cycles, pregnancy and

(*Felis pardalis*). *Journal of Reproduction and Fertility*, Vol.106, pp. 87-94 a. Swanson, W. F.; Roth, T. L.; Graham, K.; Horohov, D. W. & Godke, R. A. (1996). Kinetics of

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**13** 

*Italy* 

**Fish Spermatozoa** 

*Sciences and Technologies,* 

Loredana Zilli and Sebastiano Vilella

*University of Salento, Via Provinciale Lecce-Monteroni, Lecce* 

**Effect of Cryopreservation on Bio-Chemical** 

**Parameters, DNA Integrity, Protein Profile and** 

**Phosphorylation State of Proteins of Seawater** 

*Laboratory of Comparative Physiology, Department of Biological and Environmental* 

Fish sperm cryopreservation is considered as a valuable technique for artificial reproduction and genetic improvement (Chao & Liao, 2001; Kopeika et al., 2007; Rana, 1995; Suquet et al., 2000). Semen quality must be monitored when attempts are made to increase the efficiency of artificial fertilization, to cryopreserve only sperm of high quality, and to evaluate frozenthawed sperm. Cryopreserved sperm usually shows, with respect to fresh sperm, a lower quality, since the freezing–thawing procedure affects DNA and protein integrity (Labbe et al., 2001; Zilli et al., 2003, 2005), membrane lipids (Maldjian et al., 2005; Müller et al., 2008), sperm motility (Linhart et al., 2000; Ritar, 1999; Rodina et al., 2007; Zilli et al., 2005), fertilization ability (Gwo & Arnold, 1992; Rana, 1995), and also larval survival (Suquet et al., 1998). Spermatozoa genome alteration due to cryopreservation may affect only late embryonic development and larval survival (Kopeika et al., 2003a, 2003b, 2004; Suquet et al., 1998), but not the early events in embryonic development, because these are controlled by maternally inherited information (Braude et al., 1988). On the contrary, defects in sperm proteins (degradation and/or change of the phosphorylation state) may compromise sperm motility, fertilization ability, and the early events after fertilization (Cao et al., 2003; Huang

The most common parameters used to evaluate sperm quality are fertilization ability, motility (rate and duration) and cellular (chemical and/or biochemical) parameters. Fertilizing capacity is the most conclusive test of sperm quality but the use of this marker is laborious and requires the availability of eggs (McNiven et al., 1992). Motility is normally evaluated as percentage and duration, but some authors also use velocity, flagellum beat frequency, or other parameters measured by computer-assisted sperm analysis (Ciereszko et al., 1996; Cosson et al., 2000; Rurangwa et al., 2001). Cellular bio markers has been used to evaluate spermatozoa quality of different fish species such as Atlantic salmon (Aas et al., 1991; Hwang & Idler, 1969), rainbow trout (Ciereszko & Dabrowski, 1994; Lahnsteiner et al.,1996a, 1998) and sea bass (Zilli et al., 2004). All these

**1. Introduction** 

et al., 1999; Lessard et al., 2000).
