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World J Gastroenterol. Aug 7, 2007; 13(29): 3990-3995
Published online Aug 7, 2007. doi: 10.3748/wjg.v13.i29.3990
A novel sampling method for the investigation of gut microbiota
Alina Guimarães Quintanilha, Bruno Zilberstein, Manoel A A Santos, Denis Pajecki, Eduardo Guimarães Hourneaux Moura, Paulo Roberto Arruda Alves, Fauze Maluf-Filho, Ivan Cecconello
Alina Guimarães Quintanilha, Bruno Zilberstein, Manoel A A Santos, Denis Pajecki, Eduardo Guimarães Hourneaux Moura, Paulo Roberto Arruda Alves, Fauze Maluf-Filho, Ivan Cecconello, Digestive Surgery Division, Department of Gastroenterology, and Institute of Biomedical Sciences, Department of Microbiology, São Paulo University Medical School, São Paulo, SP, Brazil
Author contributions: All authors contributed equally to the work.
Correspondence to: Alina Guimarães Quintanilha, Digestive Surgery Division, Department of Gastroenterology, São Paulo University Medical School, São Paulo, SP,Brazil. alina.quinta@hotmail.com
Telephone: +55-11-30828000 Fax: +55-11-30818809
Received: October 2, 2006
Revised: October 15, 2006
Accepted: November 4, 2006
Published online: August 7, 2007
Abstract

AIM: In order to characterize the qualitative and quantitative microorganisms in different sites of the lower digestive tract (LDT) in healthy volunteers, a specific technique was developed for collecting mucous of the distal ileum, colon and rectum.

METHODS: A polyethylene tube was designed to go through the colonoscope channel with a No. 8 French tube. In order to avoid internal contamination, the distal extremity was protected with a membrane of microfilm after being sterilized in ethilene oxid. To facilitate the aspiration of a precise volume, its interior was coated with silicone. One hundred microlliter (0.1 mL) sample of mucous was collected and transferred into an Eppenddorff tube containing nine hundred microlliter (0.9 mL) of VMGA-3 (viable medium of Goteborg). This procedure was repeated at each site of the LDT with a new sterilized catheter.

RESULTS: All sites revealed the “non pathogenic” anaerobic bacteria Veillonella sp (average 105 colony forming units/mL-CFU/mL), allowing to conclude an environment of low oxidation-reduction potential (redox) in the LDT. It was also characterized the presence of Klebisiella sp with significant statistical predominance (SSP) in the ileum. Enterobacter sp was found with SSP in the sigmoid colon, Bacteroides sp non-pigmented (npg) and E.coli with SSP in the sigmoid colon and rectum, Enterococcus sp and Lactobacillus sp with SSP in the rectum, all in a mean concentration of 105 CFU/mL.

CONCLUSION: This procedure is feasible and efficient and can point out a similar distribution of the aerobic and anaerobic bacteria with the presence of biological markers of normal microbiota in the LDT.

Keywords: Colony count, Microbial, Fungi, Aerobic bacteria, Anaerobic bacteria, Gastrointestinal endoscopy, Large Intestine, Bacterial translocation, Microbiology, Catheterization