Cryopreservation of Nile Tilapia (Oreochromis niloticus) Sperm

Abstract

The main aim of this study is to determine the effect of the straw volume (0.25 vs. 0.5 mL) on Nile tilapia sperm quality after cryopreservation. Sperm was frozen according to conventional slow freezing procedure and diluted at ratio of 1: 3 with ionic extender containing 350 mM glucose and 30 mM Tris containing 10% dimethylacetamide. Diluted semen was equilibrated at 4 degrees C for 10 min and drawn into 0.25-mL or 0.5-mL plastic straws and sealed with polyvinyl alcohol. Samples were frozen 3 cm above of the liquid nitrogen surface and exposed to the liquid nitrogen vapor (approximate to-140 degrees C) for 10 min. After this, frozen sperm cells were kept into the liquid nitrogen container (-196 degrees C). The frozen sperm in different volume of straws were thawed in a water bath at 30 degrees C for 20 s (0.25-mL straws) or at 30 degrees C for 30 s (0.5-mL straws), respectively. Fertilization was conducted using 1 x 10(5) spermatozoa/egg ratio with each straw type. The findings of the present study indicated that cryopreservation of sperm in glucose-Tris-based extender using 0.5-mL straws improved post-thaw progressive motility, duration of progressive motility, and fertilization results (P<0.01). On the other hand, differences in term of post-thaw cell viability was not significant among the treatments (P>0.01). In conclusion, our results suggest that Nile tilapia sperm can be successfully cryopreserved in Tris-based extenders supplemented with glucose containing 10% dimethylacetamide in 0.5-mL straws.