Article
Age and disease related changes in intestinal bacterial
populations assessed by cell culture, 16S rRNA abundance, and community
cellular fatty acid profiles
M J Hopkinsa, R Sharpb, G T Macfarlanea
a Department of
Molecular and Cellular Pathology, University of Dundee, Dundee, UK, b School of Applied Science, South Bank
University, London, UK
Correspondence to: Dr M J Hopkins, MRC Microbiology and Gut Biology Group, Department of Molecular and Cellular Pathology, University of Dundee, Level 6, Ninewells Hospital and Medical School, Dundee DD1 9SY, UK m.j.hopkins{at}dundee.ac.uk
Accepted for publication 18 July 2000
BACKGROUND
The normal
intestinal microflora plays an important role in host metabolism
and provides a natural defence mechanism against invading pathogens.
Although the microbiota in adults has been extensively studied, little
is known of the changes that occur in the microflora with aging. These
may have important consequences in elderly people, many of whom are
receiving antibiotic therapy and who are most susceptible to intestinal dysbiosis.
AIMS
To characterise
the major groups of faecal bacteria in subjects of different ages using
a combination of cultural, molecular, and chemotaxonomic approaches.
METHODS
Comparative
microbiological studies were made on four different subject groups:
children (16 months to seven years, n=10), adults (21-34 years, n=7),
healthy elderly subjects (67-88 years, n=5), and geriatric patients
(68-73 years, n=4) diagnosed with Clostridium
difficile diarrhoea. Selected faecal bacteria were investigated
using viable counting procedures, 16S ribosomal RNA (rRNA) abundance
measurements, and the occurrence of specific signature fatty acids in
whole community fatty acid methyl ester profiles.
RESULTS
The principal
microbiological difference between adults and children was the
occurrence of higher numbers of enterobacteria in the latter group, as
determined by viable counts (p<0.05) and 16S rRNA (p<0.01)
measurements. Moreover, a greater proportion of children's faecal rRNA
was hybridised by the three probes (bifidobacteria, enterobacteria,
bacteroides-porphyromonas-prevotella) used in the study, indicating a
less developed gut microbiota. Species diversity was also markedly
lower in the Clostridium difficile associated diarrhoea group, which was characterised by high numbers of
facultative anaerobes and low levels of bifidobacteria and bacteroides.
Although it was a considerably less sensitive diagnostic tool, cellular
fatty acid analysis correlated with viable bacterial counts and 16S
rRNA measurements in a number of bacteria, including bacteroides.
CONCLUSIONS
Polyphasic
analysis of faecal bacteria showed that significant structural changes
occur in the microbiota with aging, and this was especially evident
with respect to putatively protective bifidobacteria. Reductions in
these organisms in the large bowel may be related to increased disease
risk in elderly people.
Keywords: intestinal bacteria; aging; cellular fatty acids; ribosomal RNA; Clostridium difficile infection
© 2001 by Gut
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