Comparison of Two Methods for Sperm Plasma Membrane Integrity Assessment in Frozen Murrah Buffalo Semen

The integrity of the sperm plasma membrane in frozen semen is crucial in assessing semen quality and is directly linked to fertility. The plasma membrane integrity of sperm can be tested using several methods, including the Hypo-Osmotic Swelling Test (HOS-Test) and Carboxyfluorescein Diacetate-Propidium Iodide Staining (CFDA-PI). This study compares the integrity of the plasma membrane of frozen Murrah buffalo semen using the HOS-Test and CFDA-PI. The semen was obtained from four buffalo bulls, each representing a different production batch. The straw was thawed individually, and quality was assessed. The plasma membrane integrity of each sample was evaluated using the HOS-Test and CFDA-PI staining methods. A two-sample t-test was used to analyze the data and demonstrate that there is a statistically significant difference in sperm plasma membrane integrity between the HOS-Test (64.83%) and CFDA-PI staining (58.97%) methods (P0.05). The HOS-Test assessment is optimal for cost-effectiveness, speed, and convenience. It can be implemented in laboratory and field settings with equal ease. In contrast, CFDA-PI staining is a good standard for accuracy and precision. It provides a comprehensive foundation for determining the quality of frozen semen.