Rabeay Hassan and Ursula Bilitewski

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Presentation transcript:

Rabeay Hassan and Ursula Bilitewski Simple Electrode System for the Detection of Viable Candida albicans and Investigations of Respiratory Pathways Rabeay Hassan and Ursula Bilitewski Helmholtz Centre for Infection Research Rabeay.hassan@helmholtz-hzi.de „Direct communication between living yeasts and electrodes“ as original title DGHM Jahrestagung 2009 - Göttingen

Objectives Utilisation of electrochemical methods for the detection of viable yeasts, in particular Candida albicans Optimisation of the experimental set-up to be able to exploit the bioelectrochemical properties of C. albicans for diagnostic purposes Proposal of electrochemical methods as a tool for studies of the structure and function of mitochondria Electrode System for the Detection of Yeasts

Experimental approach: Voltammetry Three-electrode setup: (1) Carbon Paste working electrode; (2) Platinum auxiliary electrode; (3) Ag/AgCl reference electrode Variation of current with the potential due to electron transfer Electrode System for the Detection of Yeasts

Electrochemical oxidation of yeast species C. albicans S. cerevisiae Dead cells Electrochemical conditions Supporting electrolyte is phthalate buffer, pH 7, scan rate is 50 mV/s. C. albicans, S. cerevisiae and dead cells of C. albicans at cell concentrations of 3.8x10^6, 9.15x10^6 and 2.2x10^6 /ml, respectively. Voltammetric behavior of living cells of C. albicans and S.cerevisiae Electrode System for the Detection of Yeasts

Correlation to cell numbers of C. albicans Electrode System for the Detection of Yeasts

Bioelectrochemical Mechanism: Influences of Carbon Sources The increased signal from S. cerevisiae which were cultivated with the non-fermentable carbon source galactose points to the respiratory chain as electron source. Electrode System for the Detection of Yeasts

Scheme of the C. albicans respiratory chain SHA Antimycin KCN Rotenone J Bioenerg Biomembr (2006) 38:129–135 Electrode System for the Detection of Yeasts

Influences of Respiratory Chain Inhibitors Rotenone: complex I; Antimycin: complex III; KCN: complex IV (COX) Electrode System for the Detection of Yeasts

Inhibition of the alternative oxidase (AOX) SHA; Salicylhydroxamic acid Electrode System for the Detection of Yeasts

Oxidation Current SHA Antimycin KCN Rotenone e-

Electron transfer steps in cytochrome c oxidase matrix Electrode System for the Detection of Yeasts

Deletion of genes for subunits of cytochrome c oxidase Single gene deletion mutants of S. cerevisiae deliver lower electrochemical signals, which correlate with lower oxygen consumption rates. Electrode System for the Detection of Yeasts

Conclusion C. albicans can directly transfer electrons to electrodes Electrochemical signals increase with the number of viable cells and can be used for diagnosis purposes The signal originates from the activity of cytochrome c oxidase (COX) Electrochemical systems can be used for screening of new drugs affecting the electron transport chain (ETC) Electrode System for the Detection of Yeasts

Helmholtz Centre for Infection Research ACKNOWLEDGEMENT Egyptian ministry of higher education and scientific research, and National Research Centre NRC, Cairo- Egypt Helmholtz Centre for Infection Research Electrode System for the Detection of Yeasts

Group members 2009

Thanks for your attention Rabeay Hassan

Focus on Issues How to obtain electricity from microorganisms? Where does electricity come from? Which microorganisms and what is the essential aim of this work?