Brazil currently ranks as one of the world leaders in food production and exportation. This scenario encourages the development of animal and plant health programs to ensure the production of safe food, helping the country to become an international provider of food for excellence. However, some health problems in dairy production, such as mastitis, have garnered increasing concern. This study aimed to estimate the prevalence of bovine mastitis in select properties located in the western Santa Catarina region, to assess the susceptibility profile to antimicrobial agents used for treatment and to check for the presence of genes (icaA and icaD) associated with biofilm formation in Staphylococcus aureus. In 148 milk samples collected, 72.97% had bacterial growth (n = 108). Among the isolated microorganisms, 21.62% (n = 32) were classified as Staphylococcus aureus, 18.91% (n = 28) as Staphylococcus sp. coagulase negative, 7.43% (n = 11) as Corynebacterium sp., 6.76% (n = 10) as Staphylococcus sp. coagulase positive, 5.41% (n = 8) as Nocardia sp. and 12.83% (n = 19) classified in different bacterial genera. Among the isolates submitted for antimicrobial susceptibility testing, it was observed that 8.95% (n = 6/67) had resistance to amoxicillin, 8.04% (n = 7/87) to ampicillin, 5.88% (n = 5/85) to cephalothin, 3.40% (n = 3/88) to ceftiofur and enrofloxacin, 20.45% (n = 18/88) to streptomycin, 17.04% (n = 15/88) to gentamicin and lincomycin, 31.81% (n = 28/88) to neomycin, 14.94% (n = 13/87) to penicillin and 25% (n = 22/88) to tetracycline. Staphylococcus sp. coagulase negative isolates showed higher multidrug resistance when compared to those of S. aureus and Staphylococcus sp. coagulase positive. Thirty-one strains of S. aureus isolates were genotypically tested by polymerase chain reaction (PCR), yielding a positive result for the icaA gene in 83.87% of the samples, 80.64% positive for icaD and 74.19% of these showed both genes. The results reinforce the importance of controlling this disease by adopting strict hygiene protocols and disinfection of equipment used for milking and the prescription of appropriate therapies to clinical and subclinical cases of mastitis, as well as evaluating not only the susceptibility profile of isolates but also the biofilm formation capacity. ResumoO Brasil situa-se hoje como um dos líderes mundiais em produção e exportação de alimentos. Este cenário incita a elaboração de programas de sanidade animal e vegetal que garantam a produção de alimentos seguros, contribuindo para que o país se torne um fornecedor internacional de alimentos por excelência. Entretanto, alguns problemas sanitários na produção leiteira, como a mastite, têm acarretado crescentes preocupações. O presente estudo teve como objetivo estimar a prevalência de mastite bovina em algumas propriedades localizadas na região Oeste de Santa Catarina, avaliar o perfil de suscetibilidade aos antimicrobianos empregados para tratamento e verificar a presença de genes (icaA e icaD) associados à formação de biofilmes em ...