Legionella colonization and 3D spatial location within a Pseudomonas biofilm
Legionella colonization and 3D spatial location within a Pseudomonas biofilm
Biofilms are known to be critical for Legionella settlement in engineered water systems and are often associated with Legionnaire's Disease events. One of the key features of biofilms is their heterogeneous three-dimensional structure which supports the establishment of microbial interactions and confers protection to microorganisms. This work addresses the impact of Legionella pneumophila colonization of a Pseudomonas fluorescens biofilm, as information about the interactions between Legionella and biofilm structures is scarce. It combines a set of meso- and microscale biofilm analyses (Optical Coherence Tomography, Episcopic Differential Interference Contrast coupled with Epifluorescence Microscopy and Confocal Laser Scanning Microscopy) with PNA-FISH labelled L. pneumophila to tackle the following questions: (a) does the biofilm structure change upon L. pneumophila biofilm colonization?; (b) what happens to L. pneumophila within the biofilm over time and (c) where is L. pneumophila preferentially located within the biofilm? Results showed that P. fluorescens structure did not significantly change upon L. pneumophila colonization, indicating the competitive advantage of the first colonizer. Imaging of PNA-labelled L. pneumophila showed that compared to standard culture recovery it colonized to a greater extent the 3-day-old P. fluorescens biofilms, presumably entering in VBNC state by the end of the experiment. L. pneumophila was mostly located in the bottom regions of the biofilm, which is consistent with the physiological requirements of both bacteria and confers enhanced Legionella protection against external aggressions. The present study provides an expedited methodological approach to address specific systematic laboratory studies concerning the interactions between L. pneumophila and biofilm structure that can provide, in the future, insights for public health Legionella management of water systems.
Biofilms/growth & development, Legionella pneumophila/physiology, Legionella/physiology, Microscopy, Confocal, Pseudomonas fluorescens/physiology, Tomography, Optical Coherence, Biofilm structure, Stagnation, Biofilm, Legionella pneumophila, Water systems, Legionella spatial location
Silva, Ana Rosa
325c3df6-056e-4663-a0b2-a4d16589b880
Melo, Luis F.
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Keevil, C. William
cb7de0a7-ce33-4cfa-af52-07f99e5650eb
Pereira, Ana
3ed70937-fab3-455d-9f66-d36b6da82252
22 July 2024
Silva, Ana Rosa
325c3df6-056e-4663-a0b2-a4d16589b880
Melo, Luis F.
0afab3f8-ee86-4b15-b52a-3bf8189d3968
Keevil, C. William
cb7de0a7-ce33-4cfa-af52-07f99e5650eb
Pereira, Ana
3ed70937-fab3-455d-9f66-d36b6da82252
Silva, Ana Rosa, Melo, Luis F., Keevil, C. William and Pereira, Ana
(2024)
Legionella colonization and 3D spatial location within a Pseudomonas biofilm.
Scientific Reports, 14 (1), [16781].
(doi:10.1038/s41598-024-67712-4).
Abstract
Biofilms are known to be critical for Legionella settlement in engineered water systems and are often associated with Legionnaire's Disease events. One of the key features of biofilms is their heterogeneous three-dimensional structure which supports the establishment of microbial interactions and confers protection to microorganisms. This work addresses the impact of Legionella pneumophila colonization of a Pseudomonas fluorescens biofilm, as information about the interactions between Legionella and biofilm structures is scarce. It combines a set of meso- and microscale biofilm analyses (Optical Coherence Tomography, Episcopic Differential Interference Contrast coupled with Epifluorescence Microscopy and Confocal Laser Scanning Microscopy) with PNA-FISH labelled L. pneumophila to tackle the following questions: (a) does the biofilm structure change upon L. pneumophila biofilm colonization?; (b) what happens to L. pneumophila within the biofilm over time and (c) where is L. pneumophila preferentially located within the biofilm? Results showed that P. fluorescens structure did not significantly change upon L. pneumophila colonization, indicating the competitive advantage of the first colonizer. Imaging of PNA-labelled L. pneumophila showed that compared to standard culture recovery it colonized to a greater extent the 3-day-old P. fluorescens biofilms, presumably entering in VBNC state by the end of the experiment. L. pneumophila was mostly located in the bottom regions of the biofilm, which is consistent with the physiological requirements of both bacteria and confers enhanced Legionella protection against external aggressions. The present study provides an expedited methodological approach to address specific systematic laboratory studies concerning the interactions between L. pneumophila and biofilm structure that can provide, in the future, insights for public health Legionella management of water systems.
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s41598-024-67712-4
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Accepted/In Press date: 15 July 2024
Published date: 22 July 2024
Keywords:
Biofilms/growth & development, Legionella pneumophila/physiology, Legionella/physiology, Microscopy, Confocal, Pseudomonas fluorescens/physiology, Tomography, Optical Coherence, Biofilm structure, Stagnation, Biofilm, Legionella pneumophila, Water systems, Legionella spatial location
Identifiers
Local EPrints ID: 492919
URI: http://eprints.soton.ac.uk/id/eprint/492919
ISSN: 2045-2322
PURE UUID: c686a386-1b15-41a6-9969-cbf39ea73950
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Date deposited: 20 Aug 2024 16:48
Last modified: 21 Aug 2024 01:38
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Author:
Ana Rosa Silva
Author:
Luis F. Melo
Author:
Ana Pereira
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