Genomic and Phenotypic Insights into the Probiotic Potential and Exopolysaccharide Production of Weissella cibaria MSCH1

Probiotics are increasingly being explored as alternatives for preventing intestinal infections in livestock, and Weissella species have attracted attention for their health-promoting properties.This study evaluated the probiotic functional traits, safety attributes, and exopolysaccharide (EPS) production potential of Weissella cibaria MSCH1, isolated from a Chiku sample.Phenotypic assays demonstrated that MSCH1 tolerated simulated gastric juice (pepsin, pH 3) with 64% survival after 2 h and maintained significant viability under simulated intestinal-like conditions (trypsin, pH 8) after 6 h.The strain exhibited strong auto-aggregation and notable antimicrobial activity against representative pathogens, supporting its potential for colonisation and competitive exclusion in the gastrointestinal tract.Whole-genome sequencing revealed a 2.489 Mb genome with 44.75% GC content, harbouring genes associated with carbohydrate metabolism, EPS biosynthesis, and stress responses, indicating adaptation to the gastrointestinal environment.Functional analysis highlighted a high abundance of glycosyltransferases and glycoside hydrolases, reflecting metabolic flexibility and strong EPS-producing capacity.Safety assessment confirmed the absence of transferable antibiotic resistance and virulence-associated genes, consistent with its non-hemolytic and antibiotic-susceptible phenotype.Comparative genomic analysis identified unique genes that may enhance ecological adaptability and functional diversity.Collectively, the integrated phenotypic and genomic data suggest that W. cibaria MSCH1 is a safe, metabolically versatile, and promising probiotic candidate with potential applications in animal health and functional food development.