In Vitro Effect of Elevated Ammonia and Urea Levels on Post-Thawed Bull Semen Sperm Characteristics

Elevated ammonia and urea, common byproducts of nitrogen metabolism, are increasingly found in dairy cows and may negatively impact reproductive function. However, their effects on bull sperm remain unclear. This study aimed to assess the impact of ammonia and urea at physiologically relevant concentrations on key sperm characteristics. Thawed bull semen was incubated under five treatment conditions: control (no added ammonia or urea), low urea (LU), high urea (HU), low ammonia (LA), and high ammonia (HA). Sperm motility and kinematics were assessed using Computer-Assisted Semen Analysis (CASA), and sperm viability, DNA integrity, mitochondrial membrane potential (MMP), and oxidative stress markers were evaluated using flow cytometry. HA significantly reduced MMP (<i>p</i> = 0.008) and several motility parameters, including progressive motility and velocity (VCL, VSL, ALH), compared to LA and control groups. A decrease in sperm viability was observed in the HA group compared to LA at the beginning of incubation. While HU reduced MMP (<i>p</i> = 0.002), sperm motility was not significantly affected compared to LU. No significant differences were found in DNA fragmentation or oxidative stress biomarkers between groups. These results highlight the impact of ammonia, specifically, on sperm mitochondrial function and motility, which are crucial for successful fertilization.