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fbtwitterlinkedinvimeoflicker grey 14rssslideshare1
Gyamfi-Brobbey, George (2016)
Languages: English
Types: Doctoral thesis
Subjects: UOWSAT
Diabetic foot ulcer (DFU), a major complication of both types 1 and 2 diabetes, develops in about 15–25% of people living with the disease. In Ghana, DFUs contribute to most hospital admissions (53%) among diabetics with high rates of amputation (33.3%) and death (8.8%). Diabetic foot ulcers are predisposed to infections from bacteria in the environment which normally colonise these wounds as multicellular communities called biofilms. Biofilms have been found to have increased resistance to antimicrobial agents probably due to the presence of an extracellular matrix that retards or prevents the entry of antimicrobial agents into the bacterial community, antibiotic resistance genes and/or the presence of persister cells that are unresponsive to antimicrobial agents. The work presented here studied the role of 2 multidrug resistant DFU isolates, Klebsiella pneumoniae and Proteus mirabilis in maintaining the chronicity of diabetic foot ulcers. Using 3 in vitro biofilm models; the conventional microtitre plate and Minimum Biofilm Eradication Concentration (MBEC™) High-Throughput assays and the Quasi–Vivo® continuous flow system, K. pneumoniae and P. mirabilis were found to be positive for acyl–homoserine lactone production, biofilm and persister cell producers and could resist and/or tolerate antibiotics such as ceftazidime and levofloxacin up to 1280 times their minimum inhibitory concentration. K. pneumoniae and P. mirabilis were also found to express the interspecies AI–2 quorum sensing molecules which significantly increased biofilm formation and fold induction of bioluminescence in a luxS mutant V. harveyi reference strain. Quorum sensing (QS) inhibition assays using baicalin hydrate, cinnamaldehyde and 2(5H)–furanone showed considerable inhibition of K. pneumoniae and P. mirabilis biofilm formation but failed to completely inhibit their growth. The combinatorial effects of antibiotics and QS inhibitors/antimicrobial peptides such as polymyxin B and polymyxin B nonapeptide determined as fractional inhibitory concentration (FIC) index suggests that, additive and synergistic effects produced by the combination of two antimicrobial agents have the potential to eradicate biofilms. Data from the FIC indices determined from the combination assays can provide the basis for the formulation of topical treatment for DFUs.
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    • tumefaciens NCIMB 14543 biosensor reporter strain. ........................................................181 Figure 5.11 AI-2 cross-stimulation assay for light production in V. harveyi. .....................182 Figure 5.12 Fold induction of luminescence of V. harveyi NCIMB 1280 reporter strain. ...186 Figure 5.13 Biofilm formation by V. harveyi NCIMB 1280 reference strain before and after mutagenesis and addition of exogenous AI-2. ......................................................................187 Figure 5.14 Specificity of carbohydrate-binding lectin to polysaccharides on whole cells of K. pneumoniae (ODKp), P. mirabilis (ODPm) and S. aureus (ODSa).. ....................................188 Figure 5.15 Effect of wound dressings on biofilm formation by K. pneumoniae and P.
    • mirabilis.. ...............................................................................................................................197 Figure 5.16 Combined effects of wound dressings and antibiotics ......................................198 Figure 5.17 Biofilm phenotypes of P. mirabilis in the presence of ......................................202 Figure 5.18 Combined effects of wound dressings and antibiotics. .....................................203 Figure 5.19 Combined effects of wound dressings and antibiotics. .....................................205 Gyamfi-Brobbey, G., Greenwell, P., Kimmitt, P. (2012). Diabetic foot ulcers in Ghana.
    • Poster presentation at the 2012 Postgraduate Fair, University of Westminster, London, UK Gyamfi-Brobbey, G., Greenwell, P., Kimmitt, P. (2013). Diabetic foot ulcers: prevalence of diabetic foot infections among the diabetic population attending the Komfo Anokye Teaching Hospital, Kumasi, Ghana. Poster presentation at the 2013 Postgraduate Fair, University of Westminster, London, UK Gyamfi-Brobbey, G., Greenwell, P., Kimmitt, P. (2014). The microbiology of diabetic foot ulcers: A Ghanaian perspective. Poster presentation at the 2014 Postgraduate Fair, University of Westminster, London, UK Gyamfi-Brobbey, G., Greenwell, P., Kimmitt, P. (2015). The microbiology of diabetic foot ulcers: A Ghanaian perspective. Oral and poster presentation at the 2015 Postgraduate Fair and 2015 Annual Graduate School Reception - dubbed “Next Generation Research,” University of Westminster, London, UK Omotosho, O., Gyamfi-Brobbey, G., Greenwell, P., Kimmitt, P. (2015). In vitro study of the effect of wound dressings on planktonic cells and biofilms from diabetic foot infections.
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