The effect of centrifugation, osmotic pressure variations and pipetting on drone semen quality

Drone semen undergoes various processes, including cryopreservation, genetic research and spermatological parameter analysis. Physical stress factors during these processes may negatively affect semen quality. This study evaluated the effects of osmotic pressure variations, repeated pipetting and centrifugation on drone semen, focusing on motility, plasma membrane integrity (PMI) and mitochondrial membrane potential (MMP). Exposure to different osmotic variations caused PMI loss at 200 mOsm/kg and motility losses at 200, 400 and 500 mOsm/kg. Repeated pipetting adversely impacted both motility and PMI. Centrifugation reduced motility at 800 × g and above, with PMI decreasing at 1000 × g and higher. MMP showed no statistically significant differences across treatments. Comparative analysis with other species revealed that semen from humans, canines and rats exhibited motility and PMI reductions at lower centrifugation forces. In another study conducted on drone semen, this effect was observed at forces of 1000 × g and above. Repeated pipetting decreased motility and PMI by up to 40% in rat and mouse semen and by 10% in ram, bull and boar semen. Osmotic pressure variations also adversely affected rat and ram semen, though drone semen displayed greater sensitivity. The most reliable parameters for drone semen were identified as centrifugation at 600 × g for 10 min, up to five pipetting repetitions and dilution within 250–300 mOsm/kg. These findings highlight the significant impact of stress factors on drone semen, emphasizing the need for optimized handling protocols to maintain semen integrity.