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The objective of this study was to identify and quantify bacteria from fresh ram semen, evaluate the use of sanitizer Kilol-L® prior to semen collection, and test the sensitivity of bacterial strains against antibiotics. We selected 24 Santa Inês rams clinically healthy, at 4 years of aged, and grouped them into two systems: on pasture (n=12) and confined (n=12). The microbiological results indicated that of the total of 120 ejaculates, 99 had bacterial growth, representing 82.5% of the samples. The most frequently isolated bacteria were Bacillus spp., Corynebacterium spp., Escherichia coli, Listeria spp., Staphylococcus spp., and Streptococcus spp. From the tested antibiotics, amicacin and gentamicin were 100% effective for Bacillus spp. and E. Coli. Ceftiofur was effective for all isolates except for Rodococcus equi. Streptococcus spp. were susceptible to ampicillin and erythromycin and Staphylococcus spp. were sensitive to gentamycin. The use of Kilol-L® reduced the number of Colony Forming Units (CFU/mL) from ejaculate without damaging semen quality. From the tested antibiotics, ceftiofur and gentamicin were more effective against the isolated bacterial strains; thus the use of these antibiotics in the ram semen extender is an alternative to control bacterial growt

IDENTIFICAÇÃO DE BACTÉRIAS NO SÊMEN DE OVINOS EM DIFERENTES SISTEMAS DE CRIAÇÃO E O EFEITO DO USO DE KILOL-L®