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Related Experiment Video

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Clostridioides difficile LuxS mediates inter-bacterial interactions within biofilms.

Ross T Slater1, Lucy R Frost1, Sian E Jossi1

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|July 11, 2019
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Clostridioides difficile LuxS/AI-2 influences single-species biofilm formation and inter-bacterial interactions. LuxS absence impacts C. difficile growth and Bacteroides fragilis interactions, revealing distinct community-forming mechanisms.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Biofilm Formation

Background:

  • Clostridioides difficile is an anaerobic gut pathogen forming biofilms, crucial in recurrent infections.
  • Mechanisms of C. difficile community formation and inter-bacterial interactions remain unclear.
  • AI-2, a quorum sensing molecule produced by C. difficile, affects biofilm formation in various bacteria.

Purpose of the Study:

  • Investigate the role of LuxS/AI-2 in C. difficile biofilm development.
  • Explore LuxS-mediated C. difficile interactions with other gut bacteria, specifically Bacteroides fragilis.
  • Elucidate the mechanisms underlying C. difficile community formation in single and mixed species contexts.

Main Methods:

  • In vitro biofilm assays using wild type and luxS mutant C. difficile strains.
  • Transcriptomic analysis (RNA-sequencing) of single and dual-species biofilms.
  • Co-culture experiments of C. difficile with Bacteroides fragilis.

Main Results:

  • A C. difficile luxS mutant showed impaired biofilm development in vitro.
  • Transcriptomic analysis revealed downregulated prophage loci in luxS mutant biofilms.
  • C. difficile growth was inhibited by B. fragilis in mixed biofilms, an effect alleviated by the absence of C. difficile LuxS.
  • Dual-species RNA-sequencing indicated AI-2 modulates metabolic pathways in B. fragilis during co-culture.

Conclusions:

  • C. difficile LuxS/AI-2 plays a role in single-species biofilm formation, potentially involving phage-mediated lysis.
  • AI-2 mediates inter-bacterial interactions, influencing C. difficile growth inhibition by B. fragilis.
  • LuxS/AI-2 utilizes distinct mechanisms for single and mixed-species community development in the gut microbiome.