The in vitro screening of lactobacilli isolates from uttapam batter, fermented supplementing with Piper betle L. leaves, was performed in order to select potent isolates for probiotic use. Their resistance to simulated gastric and intestinal juices as well as their adhesion to epithelial intestinal HCT-15 and vaginal HeLa cell lines were assessed and also evaluated for their immunomodulatory ability in ex vivo condition. Isolates were able to adhere as well as reduce pathogen adhesion to monolayer cell lines’ surfaces and reduced production of pro-inflammatory cytokine TNF-α in LPS treated PBMCs, while enhanced that of anti-inflammatory cytokine IL-10. The isolates exhibited properties of auto-aggregation, co-aggregation, hydrophobicity, bile salt hydrolase activity and strong antimicrobial activity against pathogenic bacteria. Thus, the different lactobacilli isolates displayed potent probiotic and immunomodulatory properties among which AJ7 and AJ82 had a great potential and may have applications in fermented foods as immunomodulatory probiotic additives.
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