Competitive exclusion cultures (CEC), which use whole bacterial communities derived directly from the gastrointestinal tract, have been effective to control of pathogens in animals, but its application in humans has not been widely studied. In this work, the inhibitory effect of supernatants obtained from a CEC developed from fecal samples of healthy individuals, was quantified over the growth of intestinal pathogens of public health significance. Baranyi Model was adjusted (R2>0.947) to growth curves of Shigella sonnei-(SF1), E. coli EPEC-(EC6) y Salmonella Enteritidis-(SE3) treated with supernatants, in order to estimate kinetic parameters. Both, growth rate and pathogen concentrations were significantly reduced by the action of supernatants in 89.89% and 86.80%, respectively, for the most sensitive bacteria (SF1) and 23.21% and 36.86%, for the most resistant (SE3). A growth delay of more than 14 hours was presented by SF1 and EC6 with the treatments. Thermal and enzymatic (proteinase K and trypsin) sensitivity of supernatants indicate that antagonism was mediated by bacteriocin-like substances. The CEC developed, have biotechnological potential for the production of antimicrobial substances of interest in the food and pharmaceutical industries.