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Applied and Environmental Microbiology, November 2005, p. 7483-7492, Vol. 71, No. 11
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.11.7483-7492.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Colonization of Mucin by Human Intestinal Bacteria and Establishment of Biofilm Communities in a Two-Stage Continuous Culture System

Sandra Macfarlane,* Emma J. Woodmansey, and George T. Macfarlane

Microbiology and Gut Biology Group, University of Dundee, Dundee, United Kingdom

Received 5 April 2005/ Accepted 23 July 2005

The human large intestine is covered with a protective mucus coating, which is heavily colonized by complex bacterial populations that are distinct from those in the gut lumen. Little is known of the composition and metabolic activities of these biofilms, although they are likely to play an important role in mucus breakdown. The aims of this study were to determine how intestinal bacteria colonize mucus and to study physiologic and enzymatic factors involved in the destruction of this glycoprotein. Colonization of mucin gels by fecal bacteria was studied in vitro, using a two-stage continuous culture system, simulating conditions of nutrient availability and limitation characteristic of the proximal (vessel 1) and distal (vessel 2) colon. The establishment of bacterial communities in mucin gels was investigated by selective culture methods, scanning electron microscopy, and confocal laser scanning microscopy, in association with fluorescently labeled 16S rRNA oligonucleotide probes. Gel samples were also taken for analysis of mucin-degrading enzymes and measurements of residual mucin sugars. Mucin gels were rapidly colonized by heterogeneous bacterial populations, especially members of the Bacteroides fragilis group, enterobacteria, and clostridia. Intestinal bacterial populations growing on mucin surfaces were shown to be phylogenetically and metabolically distinct from their planktonic counterparts.

* Corresponding author. Mailing address: Microbiology and Gut Biology Group, University of Dundee Medical School, Level 6, Ninewells Hospital and Medical School, Dundee DD1 9SY, Scotland, United Kingdom. Phone: 44 1382 632535. Fax: 44 1382 633952. E-mail: s.macfarlane{at}dundee.ac.uk.

Applied and Environmental Microbiology, November 2005, p. 7483-7492, Vol. 71, No. 11
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.11.7483-7492.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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