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The objective was to analyse the antibacterial ability of members of the intestinal microbiota of <em>Oncorhynchus mykiss</em> specimens on several bacterial target strains that contaminate food and cause disease in humans. Bacterial colonies from an intestinal portion of the 20 specimens of <em>O. mykiss</em> were obtained in different culture media. Several of the colonies showed antibacterial action on different target strains. The bacterial species with the highest antagonistic capacity were <em>Hafnia alvei</em> and <em>Lactococcus lactis</em> and the more susceptible target strains were <em>Aeromonas hydrophila</em>, <em>Listeria monocytogenes</em> and <em>Escherichia coli</em>. Moreover, it was shown that all the antibacterial substances were susceptible to the action of various proteolytic enzymes. The detection of substances of a proteinaceous nature, possibly bacteriocins produced by bacteria of the intestinal microbiota of <em>O. mykiss</em>, allows further study of these products to establish biotechnological developments in the area of health protection and food biopreservation.