Introduction: Lecithin nanoliposome (nano-LPO), with its cryoprotective properties, is considered to enhance the performance of a traditional semen cryoprotectant. Objective: To determine the optimal dose of lecithin nano-LPO added to the rooster semen extender. Materials and Methods: Semen samples collected weekly from eight broiler breeder roosters were mixed and aliquoted into five equal subsamples, during the five successive weeks. The subsamples were then diluted with a semen extender containing 0%, 0.5%, 1%, 1.5%, or 2% of lecithin nano-LPO. Post-thawed semen quality attributes, including sperm motility and velocity parameters, plasma membrane functionality, mitochondrial membrane potential (MMP), apoptosis-like changes, and fertility potential, were evaluated. Results: Total motility and velocity parameters, including curvilinear velocity (VCL), straight-line velocity (VSL), average path velocity μm/s (VAP), straightness (STR), linearity (LIN), lateral head displacement (ALH), and wobble (WOB) were quadratically (p < 0.01) influenced by graded levels of lecithin nano-LPO, such that the highest values were obtained when 1% of lecithin nano-LPO was used. Treatments had no significant effect on plasma membrane functionality; however, MMP (p < 0.08) and percentages of live and dead spermatozoa (p < 0.05) quadratically responded to increasing levels of lecithin nano-LPO, where the best outcome was found when about 1% of lecithin nano-LPO was used in the semen extender. The percentage of apoptotic spermatozoa cubically responded to increasing levels of lecithin nano-LPO (p ≤ 0.07). No significant trend of fertility rate was found in response to addition of lecithin nano-LPO levels. Conclusions: Supplementing an extender with 1.10% of lecithin nano-LPO is shown to be the optimal dose associated with the most improvement in post-thawed rooster sperm velocity measurements.
Keywords: cryopreservation; fertility; flow cytometry; rooster; semen quality; spermatozoa.