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

Michał Piotrowski,Dorota Wultańska,Piotr Obuch-Woszczatyński,Hanna Pituch

doi:10.1007/s10096-019-03635-7

Michał Piotrowski, Dorota Wultańska + Show 2 more

Open Access

https://doi.org/10.1007/s10096-019-03635-7

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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.

Highlights

Clostridium difficile (C. difficile) is a Gram-positive anaerobic spore-forming bacterium; it is the primary cause of nosocomial diarrhoea, associated with disturbance of the intestinal microbiota
The primary aim of this study was to investigate the effects that specific carbohydrates with prebiotic status and candidates for prebiotics chosen based on their varying degree of polymerization have on the ability of motile and non-motile C. difficile strains to adhere to various human epithelial cell lines and to compare the adhesive properties of specific C. difficile strains to these different cell lines
Our study assessed the adhesive capacity of motile and nonmotile C. difficile, the effects of 5 prebiotics on this adhesive effect, and the influence of chosen prebiotics (FOS and MAN) on biofilm formation

Summary

Introduction

Clostridium difficile (C. difficile) is a Gram-positive anaerobic spore-forming bacterium; it is the primary cause of nosocomial diarrhoea, associated with disturbance of the intestinal microbiota. This microorganism is an etiological agent for antibiotic-associated diarrhoea as well as several clinical complications, including pseudomembranous colitis, toxic megacolon, and intestinal perforation, which has a high mortality rate [1]. An additional binary toxin-CDT (CDTa, 48 kDa and CDTb, 74 kDa) is produced by some strains [1, 2]. The hypervirulent epidemic strains (A+B+CDT+) are classified as PCR-ribotype 027/North American Pulsotype 1 (NAP1)/restriction endonuclease analysis (REA) BI type (27/NAP1/BI) toxinotype III and are primarily associated with hospital acquired C. difficile infections (CDI) [2]. In a study that examined the causative agents of CDI in hospitals across Poland (2012–2013), RT027 strains were identified as the most prevalent PCR-ribotype [6]

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