Bacterial contamination of ram semen, antibiotic sensitivities, and effects on sperm quality during storage at 15°C

This study was designed to determine the degree and type of bacterial contamination of ejaculated semen samples in fertile rams and its consequences on sperm quality during storage. In experiment 1, 68 ejaculates from 36 rams were divided into two aliquots, one of which was used for bacterial culture, while the other one was diluted, stored at 15°C and assessed for plasma membrane integrity and motility at 0, 24 and 48h after dilution. From the 68 ejaculates, 66 were positive for aerobic bacteria, including 20 species of bacteria from 14 genera. The most frequently isolated bacteria were Escherichia coli, Proteus mirabilis, Enterobacter cloacae, Staphylococcus epidermis, and Staphylococcus aureus species. These 5 bacteria were present in 97% of all contaminated samples. All contaminant bacteria were found to be sensitive to gentamicin and to ceftiofur, with variable percentages of resistance to the other antibiotics evaluated. In samples with total enterobacteria count lower than 100 colony-forming units (CFU)/ml, higher proportions of motile and progressive sperm and higher velocities of spermatozoa were observed at different times during storage. In experiment 2, pure cultures of the most frequently isolated bacteria were individually added to fresh semen samples of low contamination and tested for their effects on sperm quality during storage at 15°C. Semen with E. coli showed a drastic reduction in motility, velocity and viability during storage. This reduction was also significant, but less drastic, in semen with E. cloacae and P. mirabilis, whereas it was partial and less pronounced in the other groups (S. epidermidis and S. aureus). In conclusion, the contamination of ram semen with enterobacterial species reduced sperm quality during storage at 15°C, and the antibiotics gentamicin and ceftiofur showed the higher antimicrobial activities.