Influence of omega‐3 incorporation in sperm preservation medium on physical and kinematic properties of chilled and cryopreserved ram spermatozoa

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Two experiments were carried out to investigate the efficiency of supplementing sperm preservation medium with omega-3 polyunsaturated fatty acids on improving liquid-chilled storage and cryopreservation capacity of ram spermatozoa. Ejaculates (n=100) were collected from five adult rams, Ovis aries, by an artificial vagina twice weekly throughout the period February-April, 2017. After initial evaluation, ejaculates of each collection session from the same males were pooled, diluted (1:10) with Tris-citric acid egg yolk extender, and were further split into five aliquots using a split-sample technique. The first aliquot served as control (omega-free), whereas the other four portions were supplemented with 0.1, 0.2, 0.3 or 0.4mM omega-3, respectively (T0 ). Thereafter, the diluted specimens were stored at 4°C for 48hr, during which sperm physical and morphometric properties were evaluated along with oxidative stress indices (T24 , T48 ). Omega-3 levels that efficiently mitigated the detrimental effects of chilled preservation, and maintained preservation aptitude of spermatozoa were further investigated for sperm cryosurvival against control (untreated). Post-thaw physical and kinematic properties of spermatozoa, in all groups, were objectively evaluated by a computer-assisted sperm analysis (CASA) system. The results showed that, at 48hr of chilled storage, supplementing preservation medium with 0.4mM omega-3 was positively correlated (p<0.01) with each of progressive motility, live sperm, intact acrosome and intact cell membrane (r=0.83, 0.85, 0.85, 0.89, respectively). Furthermore, a positive correlation (p<0.01) was observed between inclusion of omega-3 in cryopreservation medium and each of post-thaw total sperm motility, progressive motility, live sperm, normal sperm, intact acrosome, intact cell membrane, VCL, VSL, VAP, ALH and STR (r=0.76, 0.84, 0.79, 0.90, 0.89, 0.91, 0.61, 0.73, 0.65, 0.78 and 0.60, respectively). These results accentuate efficiency of supplementing the diluent with omega-3 fatty acids on improving chilled and cryopreservation aptitude of ram spermatozoa.