A Novel Recombinant Eel Pout (Macrozoarces americans) Type III Antifreeze Protein Improves Cryosurvival of Buffalo Sperm.

Background: The quality of cryopreserved buffalo semen is low due to high susceptibility of sperm membranes to cold shock.

Objective: The present study was designed to investigate the effect of recombinant type-III antifreeze protein from the eel pout Macrozoarces americanus (rAFPIII) on freezability of buffalo semen.

Materials And Methods: Semen was collected from three buffalo bulls for three weeks (replicates). Qualified ejaculates (N=18) were split into four aliquots and diluted in Tris-citric acid extender containing 0.0, 0.1, 1 and 10 µg mL of rAFPIII. Semen was cooled to 4 C, evaluated for sperm motility and PMI, cryopreserved and assessed for post-thaw quality.

Results: Supplementation of the extender with rAFPIII didn't affect motility and PMI of chilled semen. Post-thaw sperm motility and PMI were higher in extender supplemented with rAFPIII (10µg mL) compared to control. Sperm viability and acrosome integrity remained the same.

Conclusion: Addition of rAFPIII in extender improved motility and PMI of cryopreserved buffalo semen.