The ability of a newly developed competitive exclusion (CE) culture of lactic acid bacteria (LAB) consisting of Lactobacillus plantarum 42, L. acidophilus 38 and Lactococcus lactis 66 to control Salmonella colonization in broiler chicks was assessed. In the first in vivo trial, oral inoculation, spray and administration of the CE culture through the drinking water were tried. LAB and Salmonella counts in the cecal contents of birds at days 3 and 10 were determined. Salmonella colonization of the ceca of birds challenged with Salmonella serovar typhimurium was significantly lower (P<0.05) in chicks provided with the CE cultures as compared to the challenged control group. The three methods did not significantly differ in terms of efficacy in controlling the pathogen. Hence, any of these methods can be used to administer the CE culture; but spraying and administration through the drinking water are more practical in the commercial setting. The effect of single, double and triple applications of the CE culture on Salmonella and LAB counts in the ceca of Salmonella challenged chicks was evaluated in the second trial. Salmonella loads in the ceca of birds treated with the CE Culture were significantly lower (P<0.05) than the challenged control birds throughout the growing period. No significant effects were noted in Salmonella counts in broilers given additional doses of the protective culture, indicating that re-application did not provide additional benefits.