Preliminary Study to Develop Sperm Cryopreservation Technique of Endangered Rita Catfish Rita rita (Hamilton, 1822)
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Abstract
The study was carried out to characterize the fresh and cryopreserved sperm of the endangered Rita catfish, Rita rita (Hamilton, 1822) in order to develop sperm cryopreservation technique. Sperm was collected by dissecting out the testes and their quality was assessed as sperm motility 96.63±3.03%, volume 2.87±0.08 μL·g-1 of fish, density 3.89±0.55×109 cells·mL-1 and pH 7.73±0.13. Activation of sperm motility was evaluated in different concentrations of NaCl solution and highest motility (96.67±1.53%) and swimming duration (70.0±2.0 min) were achieved at 0.4% (128 mmol·kg-1). Toxicity of three cryoprotectants- DMSO, methanol and ethanol mixed with three extenders, Alsever’s solution, egg-yolk citrate and urea egg-yolk at their 5, 10, and 15% concentration was tested during 5–40 min incubation period. Cryoprotectants at 5 and 10% yielded highest motility during 5 and 10 min of incubation. Sperm processed with Alsever’s solution plus 10% DMSO produced significantly (p<0.05) highest equilibration motility (89.67±5.51%) and when cooled at 10 °C·min-1 yielded highest post-thaw motility (79.33±5.13%). Frozen sperm processed with Alsever’s solution, egg-yolk citrate and urea egg-yolk and 10% DMSO was stored in liquid nitrogen Dewar and assessed their quality by examining their post-thaw motility at weekly interval. Six-week investigation demonstrated gradual reduction of motility with the progress of storage period in all three diluents, but significantly (p<0.05) highest motility was observed from Alsever’s solution even at the end of investigated storage period.
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