STANDARDIZATION OF TWO INTRACELLULAR CRYOPROTECTANTS FOR FREEZING OF PUNJAB-2 ROOSTERS SEMEN DILUTED WITH TRIS COCONUT WATER EXTENDER

Abstract

Considering the risk of epidemic diseases, in-vitro conservation is a strategic tool to secure genetic diversity. The present study is therefore aimed at evaluating the efficacy of different concentrations of glycerol and dimethlysulfoxide (DMSO) for successful cryopreservation of poultry semen extended with Tris coconut-water extender. Tris coconut water extender was prepared, DMSO and Glycerol were supplemented in it at different initial concentrations of; glycerol (4, 5, 6, 7 and 8 VIV) and DMSO (4, 5, 6, 7, 8 and 9 VIV). Ejaculate Pooled semen from 10 roosters was collected and divided into 5 parts for glycero and 6 parts for DMSO. Semen was diluted with each extenders containing cryoprotectants at dilution ratio of I :2 (semen:extender). Diluted semen was filled and sealed into 0.25ml straws and equilibrated for 4hrs period and later subjected to vapour freezing and finally immersed in liquid nitrogen container. After a period of 48hrs semen straws samples were brought out of the container for microscopic analysis. A Total of three trials were conducted. Percentage of motile, viable and membrane integrity (M.I) of post-thawed semen was significantly (P>0.05) higher in 7% (51.67±1.6, 62.62±3.8, 50.14±1.2) compared to other treatments although was statistical similar in motility (51.67±1.6) and HOST (45.15±1.1) to 8%. DMSO Post thaw motility, viability and M.I were also significantly (P>0.05) higher in 8% (46.67±9.2, 74.54±5.9, 51.14 ±7.6%)  and 9% (50.0± 0.0%, 67.81±6.7 %, 54.93±1.1 %)  than other treatments. Although, both where statistically similar to 7% concentration in motility (40.0±0.0) and M.I (47.38±1.1). It is therefore conclusive that 7-8% glycerol concentration and 8-9% DMSO concentrations is desirable for rooster semen cryopreservation.