| Summary: | Staphylococci is an opportunistic bacterial population that is permanent in the normal flora of milk and poses a serious threat to animal and human health with some virulence factors and antibiotic-resistance genes. This study was aimed at identifying staphylococcal species isolated from raw milk and to determine hemolysis, biofilm, coagulase activities, and beta-lactam resistance. The raw milk samples were collected from the Düzce (Türkiye) region, and the study data represent a first for this region. The characterization of the bacteria was performed with MALDI-TOF MS and 16S rRNA sequence analysis. The presence of <i>coa, icaB, blaZ</i>, and <i>mecA</i> was investigated with PCR. A nitrocefin chromogenic assay was used for beta-lactamase screening. In this context, 84 staphylococci were isolated from 10 different species, and the dominant species was determined as <i>S. aureus</i> (32.14%). Although 32.14% of all staphylococci were positive for beta hemolysis, the <i>icaB</i> gene was found in 57.14%, <i>coa</i> in 46.42%, <i>mecA</i> in 15.47%, and <i>blaZ</i> in 8.33%. As a result, <i>Staphylococcus</i> spp. strains that were isolated from raw milk in this study contained some virulence factors at a high level, but also contained a relatively low level of beta-lactam resistance genes. However, considering the animal–environment–human interaction, it is considered that the current situation must be monitored constantly in terms of resistance concerns. It must not be forgotten that the development of resistance is in constant change among bacteria.
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