In vitro functional screening and in vivo toxicity assessment of potential probiotic Lactiplantibacillus plantarum BCCa32 and BCCa36

With growing consumer interest in health, probiotics are gaining prominence. This study explored the survival, adherence, safety, and therapeutic properties of two indigenous Lactiplantibacillus plantarum strains, BCCa32 (MN365072) and BCCa36 (MN368224), isolated from cattle in Bhatan village, Maharashtra, India. Lacticaseibacillus casei Shirota, which is a well-known probiotic strain with commercial applications, was employed as a control for comparison. Both isolates demonstrated higher hydrophobicity to cell surface, self-aggregation (cellular coherence), deconjugation of bile salts, and phenol tolerance than the control strain . BCCa36 showed the strongest adhesion to HT-29 cells (68.56 %). It also exhibited the best DPPH radical scavenging activity (23.9 %), while BCCa32 showed the highest glutathione peroxidase activity (1.159 mU/mg of protein), surpassing the control in both assays (2.2 % and 0.730 mU/mg of protein, respectively). BCCa36 also removed the most cholesterol (29.9 %), followed by BCCa32 (23.8 %) and the control strain (17.4 %). These findings highlight the antioxidant and cholesterol-lowering potential of both isolates. Safety was validated by the absence of hemolytic, DNAse, cytotoxic activity, and no toxicity in the in vivo brine shrimp toxicity assay. Thus, the overall results suggest that both the indigenous L.plantarum strains BCCa32 and BCCa36 have promising potential as probiotics for future applications in functional foods and therapeutic supplements. • L. plantarum BCCa32 & BCCa36 showed strong autoaggregation, hydrophobicity & HT-29 adhesion. • These indicate high gut colonization potential, which is important for facilitating the functional effects of probiotics. • Both isolates showed strong bile salt hydrolase (BSH) activity and significant in vitro cholesterol assimilation. • They also showed good enzymatic and non enzymatic antioxidant potential. • Both isolates showed no signs of toxicity in the in vivo brine shrimp ( Artemia salina) model.