Elsevier

Surgery

Volume 145, Issue 4, April 2009, Pages 435-447
Surgery

Original Communication
Bacterial population moves toward a colon-like community in the pouch after total proctocolectomy

https://doi.org/10.1016/j.surg.2008.12.003Get rights and content

Background

Colonic transformation is defined by phenotypic alterations in the ileum after total proctocolectomy. Changes in microbiota of the ileal pouch and the roles of these microbes in colonic transformation, however, have not been addressed.

Methods

A total of 151 stool samples were collected from patients with ulcerative colitis patients and an ileostomy, those with an ileal pouch, and healthy control volunteers. Bacterial DNA was extracted from stool, and the diversity of complex bacteria was assessed by terminal restriction fragment length polymorphism (T-RFLP) analysis, a novel DNA-based approach that enables us to investigate the presence of nonculturable bacteria. To determine whether ileal pouch bacterial communities shift to a more colon-like distribution, the relative abundance of terminal restriction fragments that could be classified as “colonic,” “ileal,” or “common” was investigated.

Results

Cluster analysis demonstrated that most of the ileostomy samples were categorized into Cluster I or II and that less than 10% of ileostomy samples were classified into Cluster IV. In contrast, more than 90% of control samples were grouped in Cluster IV. In further analyses, the median lifetimes of pouches in Clusters I, II, III, and IV were significantly different at 11, 56, 265, and 310 days, respectively. T-RFLP patterns of the ileal pouch were characterized by a time-dependent decrease in “ileal” and increase in a part of “colonic” fragments, which represented mainly nonculturable bacteria such as the Clostridium coccoides group.

Conclusion

T-RFLP analysis demonstrated that a time-dependent shift to a “colon-like” bacterial community, including nonculturable bacteria, in the ileal pouch after total proctocolectomy.

Section snippets

Samples

Samples were collected from 49 patients who had undergone TPC with IPAA (all for UC) and 31 healthy volunteers (Table I). Diagnosis of UC was based on a combination of clinical symptoms, endoscopic findings, and histologic examination. None of the healthy volunteers was treated with any medications. At Tohoku University Hospital, patients with UC underwent routinely a 2- or 3-step operative approach, after which the ileal pouch becomes “functional” after complete closure of the covering loop

Recovery of the fecal DNA

Adequate quantities and quality of DNA samples were obtained from watery stool samples. DNA concentrations ranged between 10.5 (±2.1) ng/mg stool in the ileostomy group, 14.8 (±2.3) ng/mg in patients with a new ileal pouch (within 2 years of ileostomy closure), and 30.1 (±7.7) ng/mg in patients with an established ileal pouch (>2 years since ileostomy closure). In contrast, DNA concentrations in the control group averaged 920.9 (±320.9) ng/mg.

T-RFLP analysis and clustering of fecal bacteria

Diversity of bacterial composition in fecal samples

Discussion

First of all, this study was intended to display the difference of fecal bacterial communities between samples from the ileostomy, ileal pouch, and colon. In a dendrogram analysis, stool samples from patients with ileostomy, with an ileal pouch within or more than 2 years after ileostomy closure, and from controls were categorized into 4 clusters. In further analyses of clustering of samples from the ileal pouch, we found that the median time of formation of a “functional” pouch after ileostomy

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