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      Phenotypic diversity within a Pseudomonas aeruginosa population infecting an adult with cystic fibrosis

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          Abstract

          Chronic airway infections caused by Pseudomonas aeruginosa contribute to the progression of pulmonary disease in individuals with cystic fibrosis (CF). In the setting of CF, within-patient adaptation of a P. aeruginosa strain generates phenotypic diversity that can complicate microbiological analysis of patient samples. We investigated within- and between- sample diversity of 34 phenotypes among 235 P. aeruginosa isolates cultured from sputum samples collected from a single CF patient over the span of one year, and assessed colony morphology as a screening tool for predicting phenotypes, including antimicrobial susceptibilities. We identified 15 distinct colony morphotypes that varied significantly in abundance both within and between sputum samples. Substantial within sample phenotypic heterogeneity was also noted in other phenotypes, with morphotypes being unreliable predictors of antimicrobial susceptibility and other phenotypes. Emergence of isolates with reduced susceptibility to β-lactams was observed during periods of clinical therapy with aztreonam. Our findings confirm that the P. aeruginosa population in chronic CF lung infections is highly dynamic, and that intra-sample phenotypic diversity is underestimated if only one or few colonies are analyzed per sample.

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          Most cited references34

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          Pseudomonas aeruginosa: all roads lead to resistance.

          Pseudomonas aeruginosa is often resistant to multiple antibiotics and consequently has joined the ranks of 'superbugs' due to its enormous capacity to engender resistance. It demonstrates decreased susceptibility to most antibiotics due to low outer membrane permeability coupled to adaptive mechanisms and can readily achieve clinical resistance. Newer research, using mutant library screens, microarray technologies and mutation frequency analysis, has identified very large collections of genes (the resistome) that when mutated lead to resistance as well as new forms of adaptive resistance that can be triggered by antibiotics themselves, in in vivo growth conditions or complex adaptations such as biofilm growth or swarming motility. Copyright © 2011 Elsevier Ltd. All rights reserved.
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            Nutritional cues control Pseudomonas aeruginosa multicellular behavior in cystic fibrosis sputum.

            The sputum (mucus) layer of the cystic fibrosis (CF) lung is a complex substrate that provides Pseudomonas aeruginosa with carbon and energy to support high-density growth during chronic colonization. Unfortunately, the CF lung sputum layer has been difficult to mimic in animal models of CF disease, and mechanistic studies of P. aeruginosa physiology during growth in CF sputum are hampered by its complexity. In this study, we performed chromatographic and enzymatic analyses of CF sputum to develop a defined, synthetic CF sputum medium (SCFM) that mimics the nutritional composition of CF sputum. Importantly, P. aeruginosa displays similar phenotypes during growth in CF sputum and in SCFM, including similar growth rates, gene expression profiles, carbon substrate preferences, and cell-cell signaling profiles. Using SCFM, we provide evidence that aromatic amino acids serve as nutritional cues that influence cell-cell signaling and antimicrobial activity of P. aeruginosa during growth in CF sputum.
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              Clinical significance of microbial infection and adaptation in cystic fibrosis.

              A select group of microorganisms inhabit the airways of individuals with cystic fibrosis. Once established within the pulmonary environment in these patients, many of these microbes adapt by altering aspects of their structure and physiology. Some of these microbes and adaptations are associated with more rapid deterioration in lung function and overall clinical status, whereas others appear to have little effect. Here we review current evidence supporting or refuting a role for the different microbes and their adaptations in contributing to poor clinical outcomes in cystic fibrosis.
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                Author and article information

                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group
                2045-2322
                05 June 2015
                2015
                : 5
                : 10932
                Affiliations
                [1 ]Department of Laboratory Medicine and Pathobiology, University of Toronto , Toronto, Canada
                [2 ]Latner Thoracic Surgery Research Laboratories, University Health Network , Toronto, Canada
                [3 ]Department of Cell & Systems Biology, University of Toronto , Toronto, Canada
                [4 ]Centre for the Analysis of Genome Evolution & Function, University of Toronto , Toronto, Canada
                [5 ]Department of Pediatric Laboratory Medicine, Division of Microbiology, The Hospital for Sick Children , Toronto, Canada
                [6 ]Department of Pediatrics, Division of Infectious Diseases, The Hospital for Sick Children , Toronto, Canada
                [7 ]Department of Medicine, Division of Respirology, St. Michael’s Hospital , Toronto, Canada
                [8 ]Laboratory Medicine Program, University Health Network , Toronto, Canada
                Author notes
                [*]

                These authors contributed equally to this work.

                Article
                srep10932
                10.1038/srep10932
                4456944
                26047320
                59806d87-3913-490f-88d9-4cf5ecf56d16
                Copyright © 2015, Macmillan Publishers Limited

                This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

                History
                : 05 February 2015
                : 05 May 2015
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