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Updated: Feb 19, 2026
Cefoperazone-treated Mouse Model of Clinically-relevant Clostridium difficile Strain R20291
Published on: December 10, 2016
Biofilm Structures in a Mono-Associated Mouse Model of Clostridium difficile Infection
Anna P Soavelomandroso1, Françoise Gaudin2, Sandra Hoys1
1EA4043, Unité Bactéries Pathogènes et Santé (UBaPS), Univ. Paris-Sud, Université Paris-Saclay, Châtenay-Malabry, France.
Abstract:
Clostridium difficile infection (CDI) is a major healthcare-associated disease with high recurrence rates. Host colonization is critical for the infectious process, both in first episodes and in recurrent disease, with biofilm formation playing a key role. The ability of C. difficile to form a biofilm on abiotic surfaces is established, but has not yet been confirmed in the intestinal tract. Here, four different isolates of C. difficile, which are in vitro biofilm producers, were studied for their ability to colonize germ-free mice. The level of colonization achieved was similar for all isolates in the different parts of the murine gastrointestinal tract, but pathogen burden was higher in the cecum and colon. Confocal laser scanning microscopy revealed that C. difficile bacteria were distributed heterogeneously over the intestinal tissue, without contact with epithelial cells. The R20291 strain, which belongs to the Ribotype 027 lineage, displayed a unique behavior compared to the other strains by forming numerous aggregates. By immunochemistry analyses, we showed that bacteria were localized inside and outside the mucus layer, irrespective of the strains tested. Most bacteria were entrapped in 3-D structures overlaying the mucus layer. For the R20291 strain, the cell-wall associated polysaccharide PS-II was detected in large amounts in the 3-D structure. As this component has been detected in the extrapolymeric matrix of in vitro C. difficile biofilms, our data suggest strongly that at least the R20291 strain is organized in the mono-associated mouse model in glycan-rich biofilm architecture, which sustainably maintains bacteria outside the mucus layer.
Insights
Clostridium difficile infection (CDI) biofilms form in the mouse gut, with the R20291 strain creating unique glycan-rich structures. This biofilm formation helps sustain C. difficile colonization outside the mucus layer.
Area of Science:
- Microbiology
- Gastroenterology
- Infectious Diseases
Background:
- Clostridium difficile infection (CDI) is a significant healthcare concern with frequent recurrences.
- Host colonization and biofilm formation are crucial for CDI pathogenesis.
- Previous studies confirmed C. difficile biofilm formation on abiotic surfaces, but intestinal tract biofilm formation remained unconfirmed.
Purpose of the Study:
- To investigate the ability of in vitro biofilm-producing C. difficile isolates to colonize germ-free mice.
- To characterize the in vivo distribution and biofilm architecture of C. difficile within the murine gastrointestinal tract.
Main Methods:
- Mono-association of germ-free mice with four different C. difficile isolates.
- Analysis of pathogen burden in various gastrointestinal tract sections.
- Confocal laser scanning microscopy to visualize bacterial distribution and host-pathogen interactions.
- Immunochemistry to identify bacterial components within the intestinal mucus layer and associated structures.
Main Results:
- All C. difficile isolates colonized the murine gastrointestinal tract, with higher pathogen burden in the cecum and colon.
- Bacteria were heterogeneously distributed, primarily within 3-D structures overlaying the mucus layer, not directly contacting epithelial cells.
- The R20291 strain (Ribotype 027) formed numerous aggregates and showed abundant cell-wall associated polysaccharide PS-II within these 3-D structures.
- PS-II is a known component of in vitro C. difficile biofilms, suggesting similar architecture in vivo.
Conclusions:
- C. difficile can form biofilm-like structures within the intestinal tract of mono-associated mice.
- The R20291 strain forms a glycan-rich biofilm architecture, distinct from other tested strains.
- This in vivo biofilm organization likely contributes to sustained C. difficile colonization by maintaining bacteria outside the mucus layer.
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