The Visualization of Biofilms in Chronic Diabetic Foot Wounds Using Routine Diagnostic Microscopy Methods

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 i...

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Main Authors: Angela Oates, Frank L. Bowling, Andrew J. M. Boulton, Philip G. Bowler, Daniel G. Metcalf, Andrew J. McBain
Format: Article
Language:English
Published: Wiley 2014-01-01
Series:Journal of Diabetes Research
Online Access:http://dx.doi.org/10.1155/2014/153586
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author Angela Oates
Frank L. Bowling
Andrew J. M. Boulton
Philip G. Bowler
Daniel G. Metcalf
Andrew J. McBain
author_facet Angela Oates
Frank L. Bowling
Andrew J. M. Boulton
Philip G. Bowler
Daniel G. Metcalf
Andrew J. McBain
author_sort Angela Oates
collection DOAJ
description 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.
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institution Kabale University
issn 2314-6745
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publishDate 2014-01-01
publisher Wiley
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series Journal of Diabetes Research
spelling doaj-art-12ee008b68b645faa12c9ce43527724c2025-02-03T05:44:52ZengWileyJournal of Diabetes Research2314-67452314-67532014-01-01201410.1155/2014/153586153586The Visualization of Biofilms in Chronic Diabetic Foot Wounds Using Routine Diagnostic Microscopy MethodsAngela Oates0Frank L. Bowling1Andrew J. M. Boulton2Philip G. Bowler3Daniel G. Metcalf4Andrew J. McBain5Manchester Pharmacy School, The University of Manchester, Oxford Road, Manchester M13 9PT, UKDepartment of Medicine Manchester Royal Infirmary, The University of Manchester, Oxford Road, Manchester M13 9PT, UKDepartment of Medicine Manchester Royal Infirmary, The University of Manchester, Oxford Road, Manchester M13 9PT, UKConvaTec Ltd., Deeside CH5 2NU, UKConvaTec Ltd., Deeside CH5 2NU, UKManchester Pharmacy School, The University of Manchester, Oxford Road, Manchester M13 9PT, UKDiabetic 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.http://dx.doi.org/10.1155/2014/153586
spellingShingle Angela Oates
Frank L. Bowling
Andrew J. M. Boulton
Philip G. Bowler
Daniel G. Metcalf
Andrew J. McBain
The Visualization of Biofilms in Chronic Diabetic Foot Wounds Using Routine Diagnostic Microscopy Methods
Journal of Diabetes Research
title The Visualization of Biofilms in Chronic Diabetic Foot Wounds Using Routine Diagnostic Microscopy Methods
title_full The Visualization of Biofilms in Chronic Diabetic Foot Wounds Using Routine Diagnostic Microscopy Methods
title_fullStr The Visualization of Biofilms in Chronic Diabetic Foot Wounds Using Routine Diagnostic Microscopy Methods
title_full_unstemmed The Visualization of Biofilms in Chronic Diabetic Foot Wounds Using Routine Diagnostic Microscopy Methods
title_short The Visualization of Biofilms in Chronic Diabetic Foot Wounds Using Routine Diagnostic Microscopy Methods
title_sort visualization of biofilms in chronic diabetic foot wounds using routine diagnostic microscopy methods
url http://dx.doi.org/10.1155/2014/153586
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