Data Sheet 1_Evaluation of probiotic properties and genome analysis of the new Pediococcus acidilactici strain 46A isolated from Chinese young adults: in vitro and in vivo studies.docx

BackgroundThe human gut microbiota plays a key role in intestinal homeostasis and inflammatory diseases, prompting the search for novel probiotic strains with strain-specific functional properties. Differences in dietary habits and cultural backgrounds represent a valuable source for isolating previously uncharacterized gut bacteria. Given the increasing use of probiotics, rigorous safety assessment, including antimicrobial resistance (AMR) evaluation, is required, and the integration of whole-genome sequencing with phenotypic characterization is recommended.
ObjectivesThis study evaluated the probiotic potential of Pediococcus acidilactici strain 46A, isolated from fecal samples of healthy Chinese student volunteers, through whole-genome analysis and in vitro and in vivo functional assessments.
MethodsStrain identification was performed by MALDI-TOF mass spectrometry and 16S rRNA gene sequencing. In vitro assays assessed tolerance to simulated gastrointestinal conditions, bile salt resistance, antimicrobial activity, and adhesion to HT-29 intestinal epithelial cells. Whole-genome sequencing was used to evaluate safety- and functionality-related genetic traits. The protective effect of P. acidilactici 46A was investigated in a dextran sulphate sodium (DSS)-induced murine model of colitis, including histological evaluation, inflammatory marker analysis, and gut microbiota profiling.
ResultsP. acidilactici 46A showed high survival under simulated gastrointestinal conditions and bile exposure, antimicrobial activity against Gram-positive and Gram-negative bacteria and yeasts, and strong adhesion to HT-29 cells compared with other tested strains. Genomic analysis confirmed the absence of virulence factors and clinically relevant antibiotic resistance genes. In DSS-treated mice, oral administration of P. acidilactici 46A significantly reduced colitis severity, preserved colonic mucosal architecture, and decreased inflammatory markers, including IL-1β and IL-6.
ConclusionPediococcus acidilactici 46A is a safe and functionally active probiotic candidate that attenuates intestinal inflammation and preserves mucosal integrity in a murine model of colitis, supporting its potential application in the prevention or adjunctive treatment of inflammatory gut disorders.