Salmonella continues to be a major threat to public health, especially with respect to strains from a poultry origin. In recent years, an increasing trend of antimicrobial resistance (AMR) in Salmonella spp. was observed due to the misuse of antibiotics. Among the approaches advised for overcoming AMR, probiotics from the Lactobacillus genus have increasingly been considered for use as effective prophylactic and therapeutic agents belonging to the indigenous microbiota. In this study, we isolated lactobacilli from the ilea and ceca of hens and broilers in order to evaluate their potential probiotic properties. Four species were identified as Limosilactobacillusreuteri (n = 22, 45.8%), Ligilactobacillussalivarius (n = 20, 41.6%), Limosilactobacillus fermentum (n = 2, 4.2%) and Lactobacillus crispatus (n = 1, 2%), while three other isolates (n = 3, 6.25%) were non-typable. Eight isolates, including Ligilactobacillussalivarius (n = 4), Limosilactobacillusreuteri (n = 2), L. crispatus (n = 1) and Lactobacillus spp. (n = 1) were chosen on the basis of their cell surface hydrophobicity and auto/co-aggregation ability for further adhesion assays using the adenocarcinoma cell line Caco-2. The adhesion rate of these strains varied from 0.53 to 10.78%. Ligilactobacillussalivarius A30/i26 and 16/c6 and Limosilactobacillus reuteri 1/c24 showed the highest adhesion capacity, and were assessed for their ability to compete in and exclude the adhesion of Salmonella to the Caco-2 cells. Interestingly, Ligilactobacillussalivarius 16/c6 was shown to significantly exclude the adhesion of the three Salmonella serotypes, S. Enteritidis, S. Infantis and S. Kentucky ST 198, to Caco-2 cells. The results of the liquid co-culture assays revealed a complete inhibition of the growth of Salmonella after 24 h. Consequently, the indigenous Ligilactobacillussalivarius 16/c6 strain shows promising potential for use as a preventive probiotic added directly to the diet for the control of the colonization of Salmonella spp. in poultry.