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July 1, 2005Journal of Clinical Microbiology888 citationsOpen Access

Spatial Organization and Composition of the Mucosal Flora in Patients with Inflammatory Bowel Disease

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ASAlexander SwidsinskiJWJeffrey S. WeberVLVera Loening‐Baucke

Key Points

  • To investigate the composition and spatial organization of the mucosal flora in patients with inflammatory bowel disease (IBD) compared to other conditions.
  • Analyzed biopsy specimens from IBD patients, irritable-bowel syndrome (IBS) patients, and healthy controls using rRNA-targeted probes.
  • Utilized nonaqueous Carnoy fixative to preserve mucosal bacteria for detection.
  • Examined concentrations of mucosal biofilm and identified key bacterial components in each group.
  • 90-95% of IBD patients had mucosal bacteria at concentrations >10^9/ml, significantly higher than 35% in healthy controls.
  • Bacteroides fragilis constituted >60% of the biofilm mass in IBD patients, compared to <15% in IBS patients.
  • Biofilm presence was observed in IBD patients treated with (5-ASA) or antibiotics, indicating that these treatments suppress but do not eliminate it.

Abstract

The composition and spatial organization of the mucosal flora in biopsy specimens from patients with inflammatory bowel disease (IBD; either Crohn's disease or ulcerative colitis), self-limiting colitis, irritable-bowel syndrome (IBS), and healthy controls were investigated by using a broad range of fluorescent bacterial group-specific rRNA-targeted oligonucleotide probes. Each group included 20 subjects. Ten patients who had IBD and who were being treated with antibiotics were also studied. Use of nonaqueous Carnoy fixative to preserve the mucus layer was crucial for detection of bacteria adherent to the mucosal surface (mucosal bacteria). No biofilm was detectable in formalin-fixed biopsy specimens. Mucosal bacteria were found at concentrations greater than 10(9)/ml in 90 to 95% of IBD patients, 95% of patients with self-limiting colitis, 65% of IBS patients, and 35% of healthy controls. The mean density of the mucosal biofilm was 2 powers higher in IBD patients than in patients with IBS or controls, and bacteria were mostly adherent. Bacteroides fragilis was responsible for >60% of the biofilm mass in patients with IBD but for only 30% of the biofilm mass in patients with self-limiting colitis and 40% of the biofilm in IBS patients but for <15% of the biofilm in IBD patients. In patients treated with (5-ASA) or antibiotics, the biofilm could be detected with 4,6-diamidino-2-phenylindole but did not hybridize with fluorescence in situ hybridization probes. A Bacteroides fragilis biofilm is the main feature of IBD. This was not previously recognized due to a lack of appropriate tissue fixation. Both 5-ASA and antibiotics suppress but do not eliminate the adherent biofilm.

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Cite This Study

Swidsinski et al. (2005) studied this question.

synapsesocial.com/papers/69d7ec4911d83f35e5ae3412https://doi.org/10.1128/jcm.43.7.3380-3389.2005
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