After oral administration, probiotic lactobacilli meet a number of protection systems in the human body, such as exposure to gastric, pancreatic, and small intestinal juices. Overcoming these detrimental barriers allows living bacteria to adhere to the intestinal epithelium and permanently colonize the gastrointestinal tract (GIT), providing health benefits to the host. Based on this, the transit tolerance of 25 candidate probiotic lactobacilli from katak, yoghurt, and white-brined and yellow cheese to simulated bile and small intestinal juices of variable pH was investigated. To establish their resistance, in vitro model systems based on modified MRS media and a longer duration of action (up to 24 h of incubation) were designed. Six of the strains studied were found to show strain-specific survival capacity with low viability in conditions simulating stomach acidity and high resistance to bile and intestinal juices. In addition, the adherence capability (autoaggregation and hydrophobicity) of the strains was determined. Obtained results allowed to select Lactobacillus strains with high survival ratios while passing through the GIT and good adherence properties, which make them suitable for the development of new probiotics.