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Applied and Environmental Microbiology, June 2001, p. 2746-2753, Vol. 67, No. 6
School of Pharmacy and Biomolecular Sciences,
University of Brighton, Moulsecoomb, Brighton BN2 4GJ, United
Kingdom
Received 21 November 2000/Accepted 22 March 2001
To cause an infection, bacteriophages must penetrate the alginate
exopolysaccharide of Pseudomonas aeruginosa to reach the bacterial surface. Despite a lack of intrinsic motility, phage were
shown to diffuse through alginate gels at alginate concentrations up to
8% (wt/vol) and to bring about a 2-log reduction in the cell numbers
in 20-day-old biofilms of P. aeruginosa. The inability of
alginate to act as a more effective diffusional barrier suggests that
phage may cause a reduction in the viscosity of the exopolysaccharide. Samples (n = 5) of commercial alginate and purified
cystic fibrosis (CF) alginate were incubated with 2 × 108 purified phage per ml for 24 h at 37°C. After
incubation the samples and controls were subjected to rheological
analysis with a Carrimed controlled stress rheometer. The viscosities
of phage-treated samples were reduced by up to 40% compared to those
of controls incubated in the absence of phage. The experiment was
repeated by using phage concentrations of 1010 and
1012 phage per ml and samples taken for analysis at
intervals up to 4 h. The results indicated that there was a time-
and concentration-dependent reduction in viscosity of up to 40%
compared to the viscosities of the controls. Commercial and purified CF
alginate samples, both phage treated and untreated, were subjected to
gel filtration chromatography by using Sephacryl High Resolution S-400
medium in order to obtain evidence of degradation. The results
demonstrated that alginate treated with phage had a lower molecular
weight than untreated alginate. The data suggest that bacteriophage
migration through P. aeruginosa biofilms may be facilitated
by a reduction in alginate viscosity brought about by enzymic
degradation and that the source of the enzyme may be the bacterial host itself.
0099-2240/01/$04.00+0 DOI: 10.1128/AEM.67.6.2746-2753.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Reduction in Exopolysaccharide Viscosity as an Aid to
Bacteriophage Penetration through Pseudomonas
aeruginosa Biofilms
*
Corresponding author. Mailing address: School of
Pharmacy and Biomolecular Sciences, University of Brighton,
Moulsecoomb, Brighton BN2 4GJ, United Kingdom. Phone: 44 (0) 1273 642082. Fax: 44 (0) 1273 679333. E-mail:
g.w.hanlon{at}brighton.ac.uk.
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