Abstract
Probiotic bacteria are proposed for prevention of Clostridium difficile associated diarrhea. The aim of this in vitro study was to evaluate the influence of five Lactobacillus plantarum strains to the survival of C. difficile reference strains (M13042 and VPI 10463) and clinical isolates (n=12) using co-culturing and micro-titre plate assay. Changes in bacterial growth were assessed over the time period of 48 h. Quantitative analysis of C. difficile population revealed that there was a significant decrease of C. difficile in co-culture compared to the control (p=0.01). Susceptibility against L. plantarum was C. difficile strain specific, while L. plantarum was not affected by the presence of C. difficile. Reference strains were more sensitive to inhibition than most of the clinical strains (M13042 strain vs eight clinical strains, p=0.03; VPI vs six clinical strains, p=0.04). Fluoroquinolone resistant C. difficile strains were less inhibited by L. plantarum than sensitive strains (p<0.05). In the micro-titre plate assay experiment the inhibition of C. difficile was not related to any particular C. difficile strains however, inhibitory activity was affected by treatment of supernatants. Supernatants of tested lactobacilli inhibited the C. difficile growth from 72% to 82% if nonneutralized (p=0.001); 43% to 68% if neutralized (p=0.003) and 92% to 99% (p=0.001) if supernatant was neutralized and heated as compared to controls.
Highlights
Clostridium difficile, an anaerobic toxigenic bacterium, causes a severe infectious colitis that leads to significant morbidity and mortality worldwide
We found that L. plantarum strains were able to inhibit the growth of C. difficile in vitro
L. plantarum is part of indigenous microbiota, but its prevalence in gut may vary in different geographical areas [22]
Summary
Clostridium difficile, an anaerobic toxigenic bacterium, causes a severe infectious colitis that leads to significant morbidity and mortality worldwide. Prerequisite for colonisation and infection by C. difficile is weakening of colonization resistance by suppression of indigenous intestinal microbiota, usually due to administration of antibiotics. Both enhanced bacterial toxins and diminished host immune response contribute to symptomatic disease. Along with conventional antibiotic therapy, administration of probiotics to manage C. difficile associated diarrhea is drawing increasingly more attention [2,3,6,7]. Since C. difficile infection develops after suppression of indigenous microbiota, restoration of colonization resistance with beneficial bacteria seems to be the most natural way for prevention and treatment of this infection
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