Author(s): Najmus Sakib Khan, Md. Rafiqul Islam Sarder, Mohammed Abdullah Al-Faroque, Md. Fazlul Awal Mollah

Abstract: This research dealt with standardization of sperm cryopreservation techniques of Labeo rohita. The concentration of sperm ranged from 8.55×109 to 8.57×109cells ml-1. Activation of sperm motility was decreased with increasing concentration of the extending media and motility was severely inhibited at 1% NaCl. The toxicity of cryoprotectant (DMSO and methanol) to sperm was tested at different concentrations (5%, 10%, 15%) and incubation time (5-35 min). Cryoprotectants with 5 and 10% concentrations produced better motility during 5 and 10 min incubation. Two extenders, Alsever’s solution and egg-yolk citrate, and two cryoprotectants, DMSO and methanol were used for cryopreservation of sperm. Alsever’s solution with 10% DMSO showed best performance producing 82.5±1.44% and 75.0±2.04% equilibration and post-thaw motility respectively. The fertilization and hatching rates were 62.0±0.80% and 45.5±1.78% for Alsever’s solution plus DMSO while those of 50.9±0.73% and 16±5.10% for egg-yolk citrate plus methanol respectively. Fresh sperm yielded 87.3±0.98% and 74.7±2.27% fertilization and hatching respectively. The breeding efficiency of cryopreserved sperm those were preserved with two diluents was compared and a significant variation was found for fertilization (P=0.001) as well as for hatching (P=0.006). The standardized cryopreservation techniques can be used to conserve the existing gene pool of rohu for cryogenic gene banking.

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