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The visualization of biofilms in chronic diabetic foot wounds using routine diagnostic microscopy methods

Oates, Angela; Bowling, Frank L.; Boulton, Andrew J. M.; Bowler, Philip G.; Metcalf, Daniel G.; McBain, Andrew J.


Angela Oates

Frank L. Bowling

Andrew J. M. Boulton

Philip G. Bowler

Daniel G. Metcalf

Andrew J. McBain


Diabetic foot wounds are commonly colonised by taxonomically diverse microbial communities and may additionally be infected with specific pathogens. Since biofilms are demonstrably less susceptible to antimicrobial agents than are planktonic bacteria, and may be present in chronic wounds, there is increasing interest in their aetiological role. In the current investigation, the presence of structured microbial assemblages in chronic diabetic foot wounds is demonstrated using several visualization methods. Debridement samples, collected from the foot wounds of diabetic patients, were histologically sectioned and examined using bright-field, fluorescence, and environmental scanning electron microscopy and assessed by quantitative differential viable counting. All samples (n = 26) harboured bioburdens in excess of 5 log10 CFU/g. Microcolonies were identified in 4/4 samples by all three microscopy methods, although bright-field and fluorescence microscopy were more effective at highlighting putative biofilm morphology than ESEM. Results in this pilot study indicate that bacterial microcolonies and putative biofilm matrix can be visualized in chronic wounds using florescence microscopy and ESEM, but also using the simple Gram stain.

Journal Article Type Article
Publication Date 2014
Journal Journal of Diabetes Research
Print ISSN 2314-6745
Electronic ISSN 2314-6753
Publisher Hindawi
Peer Reviewed Peer Reviewed
Volume 2014
Article Number 153586
APA6 Citation Oates, A., Bowling, F. L., Boulton, A. J. M., Bowler, P. G., Metcalf, D. G., & McBain, A. J. (2014). The visualization of biofilms in chronic diabetic foot wounds using routine diagnostic microscopy methods. Journal of Diabetes Research, 2014,
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Copyright Statement
Copyright © 2014 Angela Oates et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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