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Applied and Environmental Microbiology, April 2000, p. 1654-1661, Vol. 66, No. 4
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Phylogenetic Relationships of
Butyrate-Producing Bacteria from the Human Gut
Adela
Barcenilla,1
Susan E.
Pryde,1,*
Jennifer
C.
Martin,1
Sylvia H.
Duncan,1
Colin S.
Stewart,1
Colin
Henderson,2 and
Harry
J.
Flint1
Rowett Research Institute, Bucksburn,
Aberdeen AB21 9SB,1 and Robert Gordon
University, Kepplestone Campus, Aberdeen AB9
2PG,2 United Kingdom
Received 18 November 1999/Accepted 28 January 2000
Butyrate is a preferred energy source for colonic epithelial cells
and is thought to play an important role in maintaining colonic health
in humans. In order to investigate the diversity and stability of
butyrate-producing organisms of the colonic flora, anaerobic
butyrate-producing bacteria were isolated from freshly voided human
fecal samples from three healthy individuals: an infant, an adult
omnivore, and an adult vegetarian. A second isolation was performed on
the same three individuals 1 year later. Of a total of 313 bacterial
isolates, 74 produced more than 2 mM butyrate in vitro.
Butyrate-producing isolates were grouped by 16S ribosomal DNA (rDNA)
PCR-restriction fragment length polymorphism analysis. The results
indicate very little overlap between the predominant ribotypes of the
three subjects; furthermore, the flora of each individual changed
significantly between the two isolations. Complete sequences of 16S
rDNAs were determined for 24 representative strains and subjected to
phylogenetic analysis. Eighty percent of the butyrate-producing
isolates fell within the XIVa cluster of gram-positive bacteria as
defined by M. D. Collins et al. (Int. J. Syst. Bacteriol. 44:812-826, 1994) and A. Willems et al. (Int. J. Syst. Bacteriol. 46:195-199, 1996), with the most abundant group (10 of 24 or 42%) clustering with Eubacterium rectale, Eubacterium
ramulus, and Roseburia cecicola. Fifty percent of the
butyrate-producing isolates were net acetate consumers during growth,
suggesting that they employ the butyryl coenzyme A-acetyl coenzyme A
transferase pathway for butyrate production. In contrast, only 1% of
the 239 non-butyrate-producing isolates consumed acetate.
*
Corresponding author. Mailing address: Rowett Research
Institute, Greenburn Rd., Bucksburn, Aberdeen AB21 9SB, United Kingdom. Phone: 44 (0) 1224 712751. Fax: 44 (0) 1224 716687. E-mail:
sep{at}rri.sari.ac.uk.
Applied and Environmental Microbiology, April 2000, p. 1654-1661, Vol. 66, No. 4
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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