Abstract
Cryopreservation of sperm cells is an essential process applied for long-term conservation of aquatic genetic resources. The goal of this research was to determine effect different of straws volumes and thawing rates on the post-thaw quality and fertilization ability of cryopreserved common carp ( Cyprinus carpio ) sperm. In this study, semen was cryopreserved according to conventional slow freezing protocol. For this aim, the cryosolution contained 75 mM NaCl, 70 mM KCl, 2 mM CaCl 2 , 1 mM MgSO 4 and 20 mM Tris (pH: 8) supplemented with 10% MeOH. Following equilibration at +4°C for 10 min, semen was packed into 0.25, 0.5 and 1.5 mL straws and frozen in liquid nitrogen vapour (for 10 min at -120ºC) and finally stored in liquid nitrogen (-196ºC) tank. Thawing of cryopreserved semen process was performed at 30ºC for 10, 20 and 30 seconds in a water bath. Fertilization was performed using ratio of 1x10 5 spermatozoa/egg. The highest fertility (68.4±2.5%) was determined with cryopreserved sperm packed in 1.5 mL straws that thawed at 30ºC for 30 s. According to the results of this research, sperm cryopreserved with ionic extender containing 10% methanol and packed in 1.5 mL straws are suitable to achieve high fertilization of common carp eggs.
Highlights
In aquaculture, cryopreservation offers many benefits such as preservation of aquatic genetic diversity, selective breeding, hybridization and supply of gametes for hatchery production or laboratory researches. (Suquet et al 2000)
An acceptable cryoprotectant density is essential for the proper cryopreservation procedure
Methanol was tested for the common carp sperm cryopreservation process
Summary
Cryopreservation offers many benefits such as preservation of aquatic genetic diversity, selective breeding, hybridization and supply of gametes for hatchery production or laboratory researches. (Suquet et al 2000). Some important factors may change physiological structure of the semen These factors are: cryoprotectants and their concentrations, extender compositions, dilution ratios, equilibration periods and freezing/thawing rates (Glogowski et al 1999; Tekin et al 2007). These factors require development of species-specific cryopreservation protocol (Lahnsteiner et al 1996). Large straws can decrease the time required for semen packaging and thawing. It can accelerate sperm manipulations during fertilization (Cabrita et al 2001)
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