Journal Article

Influence of microbiota on the growth and gene expression of Clostridioides difficile in an in vitro coculture model

Rodriguez, Cristina, Martinez, Elisa, Berg, Noémie, Daube, Georges, Taminiau, Bernard

2024

Overview

Clostridioides difficile is an anaerobic, spore‐forming, Gram‐positive pathogenic bacterium. This study aimed to analyze the effect of two samples of healthy fecal microbiota on C. difficile gene expression and growth using an in vitro coculture model. The inner compartment was cocultured with spores of the C. difficile polymerase chain reaction (PCR)‐ribotype 078, while the outer compartment contained fecal samples from donors to mimic the microbiota (FD1 and FD2). A fecal‐free plate served as a control (CT). RNA‐Seq and quantitative PCR confirmation were performed on the inner compartment sample. Similarities in gene expression were observed in the presence of the microbiota. After 12 h, the expression of genes associated with germination, sporulation, toxin production, and growth was downregulated in the presence of the microbiota. At 24 h, in an iron‐deficient environment, C. difficile activated several genes to counteract iron deficiency. The expression of genes associated with germination and sporulation was upregulated at 24 h compared with 12 h in the presence of microbiota from donor 1 (FD1). This study confirmed previous findings that C. difficile can use ethanolamine as a primary nutrient source. To further investigate this interaction, future studies will use a simplified coculture model with an artificial bacterial consortium instead of fecal samples. The presence of microbiota did not affect the growth of Clostridioides difficile in this study. However, it did influence the expression of C. difficile genes related to sporulation, germination, and virulence, which are crucial for the transmission of the pathogen. In the presence of microbiota, C. difficile activates defence mechanisms to survive competition, such as adapting to an iron‐limited environment and utilizing ethanolamine metabolism.

• Publisher: John Wiley & Sons, Inc,Wiley,John Wiley and Sons Inc
• Subject: Antibiotics / Bacteria / Biosynthesis / Clostridioides difficile / Clostridioides difficile - genetics / Clostridioides difficile - growth & development / Coculture Techniques / Ethanolamine / Experiments / Fecal microflora / Feces / Feces - microbiology / Fermentation / gastrointestinal microbiome / Gene expression / Gene Expression Regulation, Bacterial / Genes / Genomes / Germination / Glycerol / growth / Humans / in vitro technique / Iron deficiency / Life sciences / Metabolism / Microbiologie / Microbiology / Microbiota / Microbiota - genetics / Microorganisms / Nutrient deficiency / Original / Polymerase chain reaction / Sciences du vivant / Spores / Spores, Bacterial - genetics / Spores, Bacterial - growth & development / Sporulation / Toxins