Fructooligosaccharides and mannose affect Clostridium difficile adhesion and biofilm formation in a concentration-dependent manner

Abstract

The aim of this study was to investigate the effects that prebiotic and candidates for prebiotics on Clostridium difficile strains to adhere to various human epithelial cell lines and to compare the adhesive properties of specific C. difficile strains. We also sought to examine the effect of different concentrations of fructooligosaccharides and mannose on the formation of biofilms by C. difficile strains. The influence of cellobiose, fructooligosaccharides, inulin, mannose, and raffinose on the adherence properties of various C. difficile strains, including motile 630, non-motile M120, and 10 clinical motile ribotype 027 strains, to non-mucous secreting HT-29, mucous secreting HT-29 MXT, and CCD 841 CoN cells lines. The most effective prebiotics were used in biofilm formation assays. We demonstrated that all C. difficile strains adhered to all cell lines. However, the C. difficile M120 non-motile strain was statistically more likely to adhere to all three cell lines (CFU median, 40) compared to the motile strains (CFU median, 3; p < 0.001). Furthermore, among the carbohydrates examined, only fructooligosaccharides and mannose were found to significantly decrease adhesion (p < 0.001) of C. difficile strains. Alternatively, using a biofilm assay, we observed, via confocal laser scanning microscopy, that sub-inhibitory concentrations (1%) of fructooligosaccharides and mannose functioned to increase biofilm formation by C. difficile. We demonstrated that specific prebiotics and candidate prebiotics exhibit varying anti-adhesive properties towards C. difficile in vitro and that treatment with sub-inhibitory concentrations of prebiotics can cause an increase in biofilm formation by C. difficile.

Impact of diet and the bacterial microbiome on the mucous barrier and immune disorders

“…Pathogenic and commensal bacteria persist in these locations through adaptations that enable them to directly interact with mucous membranes, with varying impacts on host health 4 . GI pathogens, including Clostridium difficile , utilize adhesion to the mucus layer to access and invade the underlying epithelia and cause disease 73 . However, commensal bacteria such as Escherichia coli and Lactobacillus rhamnosus also adhere to mucus and competitively inhibit colonization by opportunistic pathogens 74,75 …”

“…4 GI pathogens, including Clostridium difficile, utilize adhesion to the mucus layer to access and invade the underlying epithelia and cause disease. 73 However, commensal bacteria such as Escherichia coli and Lactobacillus rhamnosus also adhere to mucus and competitively inhibit colonization by opportunistic pathogens. 74,75 Similarly, mucins in the lung and oral cavity, from where bacteria migrate to the lower respiratory tract, 2 inhibit biofilm formation and downregulate toxins and other virulence factors of pathogenic taxa, such as Pseudomonas aeruginosa, preventing their outgrowth and dominance and thereby promoting community diversity.…”

The effect of berberine chloride and/or its combination with vancomycin on the growth, biofilm formation, and motility of Clostridioides difficile

“…Biofilms were visualized via confocal laser scanning microscopy (CLSM) according to methods previously described by Piotrowski et al [22]. Only two strains, C. difficile 630 and clinical strain 4, were selected for the visualization of biofilm under CLSM, as they showed the highest vancomycin minimal inhibitory concentration (MIC = 4.0 mg/L) among the strains tested.…”

Wolf in Sheep’s Clothing: Clostridioides difficile Biofilm as a Reservoir for Recurrent Infections

“…Although glucose is often used to support biofilm formation, other carbohydrates can induce biofilm formation as well. Indeed, mannose and fructooligosaccharides, which are toxic at high concentrations, were able to induce biofilm formation in C. difficile at sub-inhibitory concentrations [ 248 ]. In the gut, C. difficile would use mucin-derived sugars such as N-acetylglucosamine and sialic acid.…”