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Applied and Environmental Microbiology, July 2005, p. 3692-3700, Vol. 71, No. 7
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.7.3692-3700.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

pH and Peptide Supply Can Radically Alter Bacterial Populations and Short-Chain Fatty Acid Ratios within Microbial Communities from the Human Colon

Alan W. Walker,1 Sylvia H. Duncan,1 E. Carol McWilliam Leitch,1 Matthew W. Child,2 and Harry J. Flint1*

Microbial Genetics Group, Rowett Research Institute, Greenburn Road, Bucksburn, Aberdeen, United Kingdom,1 Microbiology and Gut Biology Group, University of Dundee, Dundee, United Kingdom2

Received 23 November 2004/ Accepted 31 January 2005

The effects of changes in the gut environment upon the human colonic microbiota are poorly understood. The response of human fecal microbial communities from two donors to alterations in pH (5.5 or 6.5) and peptides (0.6 or 0.1%) was studied here in anaerobic continuous cultures supplied with a mixed carbohydrate source. Final butyrate concentrations were markedly higher at pH 5.5 (0.6% peptide mean, 24.9 mM; 0.1% peptide mean, 13.8 mM) than at pH 6.5 (0.6% peptide mean, 5.3 mM; 0.1% peptide mean, 7.6 mM). At pH 5.5 and 0.6% peptide input, a high butyrate production coincided with decreasing acetate concentrations. The highest propionate concentrations (mean, 20.6 mM) occurred at pH 6.5 and 0.6% peptide input. In parallel, major bacterial groups were monitored by using fluorescence in situ hybridization with a panel of specific 16S rRNA probes. Bacteroides levels increased from ca. 20 to 75% of total eubacteria after a shift from pH 5.5 to 6.5, at 0.6% peptide, coinciding with high propionate formation. Conversely, populations of the butyrate-producing Roseburia group were highest (11 to 19%) at pH 5.5 but fell at pH 6.5, a finding that correlates with butyrate formation. When tested in batch culture, three Bacteroides species grew well at pH 6.7 but poorly at pH 5.5, which is consistent with the behavior observed for the mixed community. Two Roseburia isolates grew equally well at pH 6.7 and 5.5. These findings suggest that a lowering of pH resulting from substrate fermentation in the colon may boost butyrate production and populations of butyrate-producing bacteria, while at the same time curtailing the growth of Bacteroides spp.

* Corresponding author. Mailing address: Microbial Genetics Group, Rowett Research Institute, Greenburn Rd., Bucksburn, Aberdeen AB21 9SB, United Kingdom. Phone: (44) 1224-716651. Fax: 1224-716687. E-mail: hjf{at}rri.sari.ac.uk.

Applied and Environmental Microbiology, July 2005, p. 3692-3700, Vol. 71, No. 7
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.7.3692-3700.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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