Evaluation of probiotic potential, safety assessment and whole genome sequencing of

Introduction: Toddy, an alcoholic drink produced through natural fermentation, is a significant reservoir of probiotic microorganisms. Probiotics are increasingly studied for their beneficial effects in managing gastrointestinal disorders, enhancing immune responses, and promoting overall health. Methods: Twenty bacterial strains were isolated from toddy samples and subjected to preliminary biochemical screening. Of these, 17 isolates were selected for further screening of antibacterial activity against foodborne pathogens. These isolates were also evaluated for probiotic traits such as auto-aggregation, hydrophobicity, tolerance to bile, acid, and NaCl. Antibiotic susceptibility was assessed against nine antibiotics. The most promising isolate was subjected to whole-genome sequencing, followed by Results: Among the 17 selected isolates, VITVTD-3, VITVTO-5, and VITVH-5 exhibited inhibitory effects against all tested food-borne pathogens. These isolates also showed moderate levels of auto-aggregation and hydrophobicity, along with the ability to withstand bile salts, acidic pH, and high salt concentrations. Antibiotic profiling revealed resistance to vancomycin in all three isolates. Antioxidant and haemolytic activity assays further supported their probiotic potential. Of these, VITVTD-3 demonstrated the most favourable combination of probiotic attributes. Whole-genome sequencing revealed that this isolate corresponds to Discussion: The findings confirm that