The objective of this study was to detect and identify the autochthonous microbiota of raw milk with antagonistic activity on Listeria monocytogenes and Salmonella Enteritidis. Three hundred sixty colonies isolated from 15 raw milk samples were tested for antagonistic activity for L. monocytogenes and S. Enteritidis using the ""spot-on-the-lawn"" method. The colonies detected as antagonistic were identified using API 20 Strep. Two types of inhibition were observed: total, characterized by the formation of a well-defined halo around the colony, and partial, with the formation of a diffused halo. Ninety-one (25.3%) colonies presented antagonistic activity for L. monocytogenes and 33 (9.2%) for S. Enteritidis. Most of the antagonistic cultures were lactic acid bacteria, mainly Lactococcus lactis ssp. lactis and Enterococcus faecium. The results indicate that microorganisms in the natural microbiota of raw milk may play an important role in the inhibition of key pathogens in dairy products.