Autochthonous Lactobacillus paracasei subsp. paracasei strains isolated from Caciocavallo cheese: identification and in-vitro investigation on potential probiotic and functional properties

Background: In recent years consumers, due to the growing interest for functional foods, have put particularly a ention on prod- ucts enriched with probiotic lactic acid bacteria for their ability to give human bene t by their assumption. Microorganisms to be considered as probiotic should be able to survive to gastrointestinal juices and colonize the intestinal tract to provide bene ts to human health. The aim of the study consisted on the in vitro investigation of novel LAB strains, isolated during cheesemaking and ripening of Caciocavallo cheese, and on their probiotic potential for food application. Methods: In order to select potential probiotic bacteria, LAB isolates were preliminarily screened for their survival to simulated gastro-intestinal transitand assayed for others activities of probiotic and functional interest. Thanks to promising a itude as poten- tial probiotics, four strains were tested to evaluate their adhesion ability on Caco-2 cell lines used as intestinal cell model. Microbiological data were analysed by one-way analysis of variance (ANOVA). Comparisons of means were carried out using post-hoc Tukey’s test. For vitro adhesion experiments, statistical signi cance was evaluated by Kruskal-Wallis test for equal medi- ans. Results: Twenty- ve LAB, presumptively identi ed as Lactobacillus spp., were isolated from cheese during 60 days of ripening and screened for their survival to simulated gastrointestinal transit. Sixteen isolates that showed a survival rate ≥85% were identi edas 10 di erent pro les of Lb. paracasei subsp. paracasei. The 10 strains tolerated high concentration of bile salts by Minimal Inhibitory Concentration and growth rate assays and showed susceptibility or moderate susceptibility against antibiotics of human and vet- erinary importance, except for the resistanceto the class of aminoglycoside antibiotics. Eight out 10 strains showed in vitro choles- terol-lowering ability, whereas all strains showed antioxidant activity of their cell-free supernatants. Moreover, the four strains with highest survival to simulated gastrointestinal transit showed the ability to adhere to Caco-2 cells. Conclusion: The results suggest that some strains may be e ective probiotics to be use as tool to design probiotic dairy products after con rmation probiotic activities in further in vivo studies. Findings of the present study suggest that four strains showed good or strong adherent ability on Caco-2 cell monolayer, that is one of prerequisite that probiotic bacteria must have to perform their functional properties.