A novel targeted/untargeted GC-Orbitrap metabolomics methodology applied to Candida albicans and Staphylococcus aureus biofilms

Weidt, S., Haggarty, J., Kean, R., Cojocariu, C. I., Silcock, P. J., Rajendran, R., Ramage, G. and Burgess, K. E. V. (2016) A novel targeted/untargeted GC-Orbitrap metabolomics methodology applied to Candida albicans and Staphylococcus aureus biofilms. Metabolomics, 12, 189. (doi:10.1007/s11306-016-1134-2) (PMID:28003796) (PMCID:PMC5097782)

Weidt, S., Haggarty, J., Kean, R., Cojocariu, C. I., Silcock, P. J., Rajendran, R., Ramage, G. and Burgess, K. E. V. (2016) A novel targeted/untargeted GC-Orbitrap metabolomics methodology applied to Candida albicans and Staphylococcus aureus biofilms. Metabolomics, 12, 189. (doi:10.1007/s11306-016-1134-2) (PMID:28003796) (PMCID:PMC5097782)

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Abstract

Introduction: Combined infections from Candida albicans and Staphylococcus aureus are a leading cause of death in the developed world. Evidence suggests that Candida enhances the virulence of Staphylococcus—hyphae penetrate through tissue barriers, while S. aureus tightly associates with the hyphae to obtain entry to the host organism. Indeed, in a biofilm state, C. albicans enhances the antimicrobial resistance characteristics of S. aureus. The association of these microorganisms is also associated with significantly increased morbidity and mortality. Due to this tight association we hypothesised that metabolic effects were also in evidence. Objectives: To explore the interaction, we used a novel GC-Orbitrap-based mass spectrometer, the Q Exactive GC, which combines the high peak capacity and chromatographic resolution of gas chromatography with the sub-ppm mass accuracy of an Orbitrap system. This allows the capability to leverage the widely available electron ionisation libraries for untargeted applications, along with expanding accurate mass libraries and targeted matches based around authentic standards. Methods: Optimised C. albicans and S. aureus mono- and co-cultured biofilms were analysed using the new instrument in addition to the fresh and spent bacterial growth media. Results: The targeted analysis experiment was based around 36 sugars and sugar phosphates, 22 amino acids and five organic acids. Untargeted analysis resulted in the detection of 465 features from fresh and spent medium and 405 from biofilm samples. Three significantly changing compounds that matched to high scoring library fragment patterns were chosen for validation. Conclusion: Evaluation of the results demonstrates that the Q Exactive GC is suitable for metabolomics analysis using a targeted/untargeted methodology. Many of the results were as expected: e.g. rapid consumption of glucose and fructose from the medium regardless of the cell type. Modulation of sugar-phosphate levels also suggest that the pentose phosphate pathway could be enhanced in the cells from co-cultured biofilms. Untargeted metabolomics results suggested significant production of cell-wall biosynthesis components and the consumption of non-proteinaceous amino-acids.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Burgess, Dr Karl and Kean, Mr Ryan and Weidt, Dr Stefan and Haggarty, Ms Jennifer and Ramage, Professor Gordon and Rajendran, Dr Ranjith
Authors: Weidt, S., Haggarty, J., Kean, R., Cojocariu, C. I., Silcock, P. J., Rajendran, R., Ramage, G., and Burgess, K. E. V.
College/School:College of Medical Veterinary and Life Sciences
College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
College of Medical Veterinary and Life Sciences > School of Medicine, Dentistry & Nursing > Dental School
College of Medical Veterinary and Life Sciences > School of Veterinary Medicine
Journal Name:Metabolomics
Publisher:Springer-Verlag
ISSN:1573-3882
ISSN (Online):1573-3890
Published Online:05 November 2016
Copyright Holders:Copyright © 2016 The Authors
First Published:First published in Metabolomics 12:189
Publisher Policy:Reproduced under a Creative Commons License

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