Supernatants of the Probiotic Bacterial Cultures at sub-MIC Levels Attenuate Virulence of Pathogenic Bacteria Towards the Model Host Caenorhabditis elegans.

Abstract

This study assessed the anti-pathogenic potential of culture supernatants from three probiotic strains viz. Lactobacillus plantarum, Bifidobacterium bifidum, and Enterococcus faecium against five different gram-positive and gram-negative pathogenic bacteria. The effect of probiotic culture supernatants on growth and production of quorum sensing (QS)-regulated pigments was assessed through broth dilution assay. In vivo anti-infective efficacy of these supernatants was evaluated by challenging the model nematode host Caenorhabditis elegans with pathogenic bacteria pre-treated with probiotic culture supernatants, and worm survival was monitored over a period of 8 days. All the probiotic culture supernatants (at 5-10 %v/v) exhibited bactericidal activity against four of the test pathogens (including three multi-drug resistant gram-negative bacteria) and they (at ≥ 1%v/v) could also modulate QS in pathogenic bacteria. Pre-treatment with culture supernatants from all the three probiotic strains were able to attenuate virulence of four of the test pathogens towards C. elegans. Worms challenged with supernatant-pre-treated pathogens recorded 75-100% better survival than those challenged with pathogens receiving no such pretreatment. Virulence-attenuating effect of the culture supernatants was also carried forward to daughter populations of the supernatant-treated parent pathogens. Lactic acid seemed to contribute notably towards anti-virulence activity of the culture supernatants, but acidic pH did not seem to be important for such activity. Culture supernatants of probiotic strains used in this study can be said to possess appreciable virulence-attenuating potential. Further investigation for characterization of these supernatants and identification of the bioactive constituents is warranted. Elucidation of the molecular basis associated with the anti-pathogenic potential of these supernatants may lead to the identification of novel targets in pathogenic bacterial populations.