In an intestinal system with a balanced microbial diversity, lactic acid bacteria (LAB) are the key element which prevents the colonization and invasion of gut pathogens. Adhesion ability is important for the colonization and competition abilities of LAB. The aim of this study was to determine the adhesion and competition abilities of LAB by using a whole-tissue model. Indigenous strains were isolated from spontaneously fermented foods like cheese and pickles. The aggregation and competition abilities of the isolates were determined, as well as their resistance to gastrointestinal conditions. Four Lactobacillus strains and one Weissella strain were found to be highly competitive against three major gut pathogens, namely Clostridium difficile, Listeria monocytogenes and Salmonella Enteritidis. Tested strains decreased the number of pathogens to below their disease-causing levels. According to the results, the numbers of C. difficile and L. monocytogenes bacteria decreased by an average of 3 log, and their adhesion rates decreased by approximately 50%. However, the number of S. Enteritidis bacteria was decreased by only 1 log compared with its initial number. We thought that the weak effect on Salmonella was due to its possession of many virulence factors. The results showed that natural isolates from sources other than human specimens like the Weissella strain in this study were quite competent when compared with the human isolates in terms of their adhesion to intestines and resistance to gastrointestinal tract conditions. It was also revealed that a whole-tissue model with all-natural layers can be successfully used in adhesion and competition tests.