Comprehensive In Vitro, Genomic, Microbiota, and Subacute Toxicological Safety Characterization of...

Lactiplantibacillus plantarum is a widely studied probiotic species; however, probiotic characteristics and safety profiles are strain-specific, requiring independent evaluation. This study characterized Lactiplantibacillus plantarum ATA-LPC98052 as a candidate probiotic raw material.

ATA-LPC98052 was evaluated for hemolysis, acid/bile tolerance, Caco-2 adhesion, cytotoxicity, and storage stability. Subacute oral safety was assessed in Wistar rats by gavage for 28 days at 1.2 × 10¹¹ CFU/kg/day; the study design incorporated selected principles of OECD Test Guideline 407. Clinical, hematological, biochemical, organ-weight, and macroscopic endpoints were evaluated. Fecal microbiota was analyzed by 16S rRNA sequencing; WGS was used for taxonomic confirmation and genomic safety screening.

ATA-LPC98052 was γ-hemolytic and maintained 60% viability at pH 1.5 and 96% at pH 5.0, while viability ranged from 74% to 82% across 0.1-0.5% bile salt concentrations. Caco-2 cell viability was 99%, adhesion exceeded 90%, and the lyophilized preparation remained stable for 15 months, maintaining 9.6 log₁₀ CFU/g. Repeated oral administration caused no mortality or consistent treatment-related toxicological pattern. Longitudinal microbiota analysis showed no significant treatment × time effects on alpha diversity or Bray-Curtis community structure, and no genus-level MaAsLin2 association was FDR-significant. WGS confirmed L. plantarum identity and no contamination; ResFinder detected no acquired antimicrobial resistance determinants meeting specified thresholds, whereas CARD/RGI identified low-identity qacJ and vanY homologs requiring cautious interpretation.

ATA-LPC98052 demonstrated favorable in vitro probiotic characteristics, technological stability, gut microbiota-modulating potential, and a favorable subacute safety profile under the tested conditions; however, microbiota findings were exploratory, and phenotypic MIC testing remains warranted.