Objective. To assess the microbiological profile in seven Boar Studs (BS) in Southern Brazil, as well as evaluate antimicrobial susceptibility response to most commonly found microorganisms in BS. Materials and methods. Bacteriologic analysis was carried out in samples from the water purification system, semen extender, raw and stored semen, lab benches, and other working surfaces. Results. Growth of a mixed bacterial population was observed in water samples from all but one BS. Approximately 85% of the BS had significant contamination on their working surfaces with at least one bacterial contaminant. A total of 86% of raw semen samples were contaminated with one or more different bacteria, while 100% of the boar studs provided contaminated samples. Bacterial susceptibility to antimicrobial agents varied from over 80% for gentamycin, neomycin and ceftiofur to 40% or less for penicillin and lincomycin. Conclusions. The identification of the critical points provides necessary support to devise better strategies to minimize contamination in BS. Also, assessing the level of antimicrobial drug resistance offers accurate information to formulate more efficient antibacterial protocols that closely observe the rational use of antibiotics.