Calcium affects stallion spermatozoa parameters in different incubation temperatures

This study was aimed to determine the effect of CaCl₂ on the motility and viability of stallion spermatozoa during different incubation temperatures. Experimental samples were prepared by diluting the ejaculates (n = 10) from three uniformly housed and fed breeding stallions with six different concentrations of CaCl₂ (A: 0.1125, B: 0.225, C: 0.45, D: 0.938, E: 1.25, and F: 1.875 mg/mL). The control samples (CON) were prepared by diluting ejaculate only with physiological solution. Samples were divided into two aliquots for analyses at different storage temperatures (5 °C and 37 °C). The motility parameters were analysed by Computer Assisted Semen Analysis system at several time intervals (0, 1, 2 and 3 h) and the viability was assessed using a mitochondrial toxicity test (MTT) realized at the end of incubation at both temperatures. Addition of CaCl₂ to stallion semen showed significant effect on motility parameters, especially in the highest concentrations at 5 °C. Significant objectionable effect of CaCl₂ on both total and progressive motility was observed at temperature 37 °C compared to control sample. However, results of velocity curved line in samples C, D and F at time 1 h and also at time 2 h in sample F showed significant positive effect of CaCl₂. Sperm viability in experimental samples did not show a significant difference compared to the control at either 5 °C or 37 °C. The results of this study did not confirm essential effect of calcium on reproductive parameters of stallion. To conclude, our study demonstrated that the effect of CaCl₂ on stallion sperm motility differs in a dose-dependent manner; however, the overall impact on motility parameters does not seem to be beneficial.