Cryopreservation of silver barb Puntius gonionotus (Bleeker) spermatozoa: effect of extender composition, cryoprotective agents and freezing rate on their postthawing fertilization ability
2008
Routray, Padmanav | Dash, Surjya Narayan | Dash, Chidananda | Swain, Priyabrat | Sarkar, Sampad Kumar | Sarangi, Niranjan
The effects of extender composition, cryoprotectant concentration and freezing and thawing on the fertilization efficiency of cryopreserved spermatozoa of Puntius gonionotus were evaluated. Computer-aided motility analysis of semen was conducted to check the suitability of spermatozoa for cryopreservation after mixing with different extenders and cryoprotective agents (CPAs). Extender-4 with an osmolality 260 mOsmol kg⁻¹and pH 7.6 was used for the cryopreservation study. Among the CPAs, dimethyl sulphoxide (DMSO) was least toxic and more than 60% fertilization was achieved when used at 1.4 M at 0 °C for 10 and 30 min, whereas the toxicity of all CPAs to spermatozoa was evident when tested at 30 °C. Semen frozen at -16 °C min⁻¹ with 1.4 M DMSO showed 70% fertilization, which was significantly higher (P<0.05) than other freezing rates. Samples thawed at 35 °C water showed a fertilization rate comparable with that of fresh semen. Computer-assisted semen analysis of fresh and frozen semen after thawing showed variations in different types of motility in spermatozoa and in their class. There was no significant difference in motility before or after cryopreservation; however, significant differences could be observed in the average path velocity (VAP), straight line velocity (VSL) and curve linear velocity (VCL). Semen of silver barb could be cryopreserved with extender-4 by addition of 1.4 M DMSO to a final cryopreservation medium (MED 2) cooled at a rate of -16 °C min⁻¹, stored in liquid nitrogen (-196 °C) and utilized after thawing at 35±2 °C.
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