Cryopreservation of Indian Red Jungle Fowl ( Gallus Gallus Murghi ) Spermatozoa: A First Approach

Abstract

The experiment was designed to show the capacity of Indian Red Jungle Fowl (Gallus gallus murghi) sperm to be cryopreserved. The changes in characteristics of Indian Red Jungle Fowl spermatozoa during different phases of cryopreservation with the glycerol cryoprotectant and straw packaging were recorded. Semen from eight mature cocks were pooled, diluted at 37 °C in a Tselutin poultry extender, cooled to 4 °C in 2 h (0.275 min⁻¹) and equilibrated for 10 min after the addition of 11% glycerol. Cooled semen was filled in 0.5 mL French straws, kept over liquid nitrogen vapour for 10 min and plunged into liquid nitrogen. Frozen semen was thawed at 37 °C for 30 s. Sperm motility, plasma membrane plasticity, livability, and acrosomal integrity were assessed at the stage of pre-dilution (37 °C), post-dilution, cooling, equilibration and freeze-thawing. The experiment was repeated five times. Sperm motility did not change at post-dilution and cooling and averaged 83.3±3.1%. However, motility declined (P<0.05) to 66.0±2.4% and 45.0±6.5% at post-equilibration and thawing stages, respectively. The sperm plasma membrane plasticity at the pre-dilution stage was 74.8±1.4% and decreased (P<0.05) to 62.3±1.7%, 51.7±1.7%, and 42.0±1.2% at the stages of post-dilution, cooling and freeze-thawing, respectively. A decline (P < 0.05) in acrosomal integrity was observed at post-cooling (83.8±2.4%), equilibration (80.8±0.8%) and thawing (73.8±2.4%). It is concluded that Indian Red Jungle sperm seem to adapt quite well to the freeze-thaw process that mainly alters the motility capacities and much less the acrosomal integrity. To our knowledge, this is the first ever report on cryopreservation of Indian Red Jungle Fowl spermatozoa.