Unique growth and morphology properties of Clade 5 Clostridioides difficile strains revealed by single-cell time-lapse microscopy.

Clostridioides difficile is a gastrointestinal pathogen of both humans and agricultural animals and thus a major One Health threat. The C. difficile species consists of five main clades, with Clade 5 currently undergoing speciation from Clades 1-4. Since Clade 5 strains are highly prevalent in agric...

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Main Authors: John W Ribis, César Nieto, Nicholas V DiBenedetto, Anchal Mehra, Pola Kuhn, Qiwen Dong, Irene Nagawa, Imane El Meouche, Bree B Aldridge, Mary J Dunlop, Rita Tamayo, Abhyudai Singh, Aimee Shen
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2025-05-01
Series:PLoS Pathogens
Online Access:https://doi.org/10.1371/journal.ppat.1013155
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author John W Ribis
César Nieto
Nicholas V DiBenedetto
Anchal Mehra
Pola Kuhn
Qiwen Dong
Irene Nagawa
Imane El Meouche
Bree B Aldridge
Mary J Dunlop
Rita Tamayo
Abhyudai Singh
Aimee Shen
author_facet John W Ribis
César Nieto
Nicholas V DiBenedetto
Anchal Mehra
Pola Kuhn
Qiwen Dong
Irene Nagawa
Imane El Meouche
Bree B Aldridge
Mary J Dunlop
Rita Tamayo
Abhyudai Singh
Aimee Shen
author_sort John W Ribis
collection DOAJ
description Clostridioides difficile is a gastrointestinal pathogen of both humans and agricultural animals and thus a major One Health threat. The C. difficile species consists of five main clades, with Clade 5 currently undergoing speciation from Clades 1-4. Since Clade 5 strains are highly prevalent in agricultural animals and a frequent cause of zoonotic infections, these strains may have evolved phenotypes that distinguish them from Clade 1-4 strains. Here, we compare the growth properties of Clade 5 strains to those of Clade 1-4 strains using anaerobic time-lapse microscopy coupled with automated image analysis. Our analyses indicate that Clade 5 strains grow faster and are more likely to form long chains of cells than Clade 1-4 strains. Using comparative genomic and CRISPRi analyses, we show that the chaining phenotype of Clade 5 strains is driven by the orientation of the invertible cmr switch sequence, with chaining strains exhibiting a bias to the cmr-ON state. Interestingly, Clade 5 strains with a bias towards the cmr-ON state shifted to a largely cmr-OFF state during murine infection, suggesting that the cmr-OFF state is under positive selection during infection. Collectively, our data reveal that Clade 5 strains have distinct growth properties, which may allow them to inhabit diverse ecological niches.
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institution Kabale University
issn 1553-7366
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language English
publishDate 2025-05-01
publisher Public Library of Science (PLoS)
record_format Article
series PLoS Pathogens
spelling doaj-art-69a3099835a3470e97f1e9c5867766672025-08-20T03:46:25ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742025-05-01215e101315510.1371/journal.ppat.1013155Unique growth and morphology properties of Clade 5 Clostridioides difficile strains revealed by single-cell time-lapse microscopy.John W RibisCésar NietoNicholas V DiBenedettoAnchal MehraPola KuhnQiwen DongIrene NagawaImane El MeoucheBree B AldridgeMary J DunlopRita TamayoAbhyudai SinghAimee ShenClostridioides difficile is a gastrointestinal pathogen of both humans and agricultural animals and thus a major One Health threat. The C. difficile species consists of five main clades, with Clade 5 currently undergoing speciation from Clades 1-4. Since Clade 5 strains are highly prevalent in agricultural animals and a frequent cause of zoonotic infections, these strains may have evolved phenotypes that distinguish them from Clade 1-4 strains. Here, we compare the growth properties of Clade 5 strains to those of Clade 1-4 strains using anaerobic time-lapse microscopy coupled with automated image analysis. Our analyses indicate that Clade 5 strains grow faster and are more likely to form long chains of cells than Clade 1-4 strains. Using comparative genomic and CRISPRi analyses, we show that the chaining phenotype of Clade 5 strains is driven by the orientation of the invertible cmr switch sequence, with chaining strains exhibiting a bias to the cmr-ON state. Interestingly, Clade 5 strains with a bias towards the cmr-ON state shifted to a largely cmr-OFF state during murine infection, suggesting that the cmr-OFF state is under positive selection during infection. Collectively, our data reveal that Clade 5 strains have distinct growth properties, which may allow them to inhabit diverse ecological niches.https://doi.org/10.1371/journal.ppat.1013155
spellingShingle John W Ribis
César Nieto
Nicholas V DiBenedetto
Anchal Mehra
Pola Kuhn
Qiwen Dong
Irene Nagawa
Imane El Meouche
Bree B Aldridge
Mary J Dunlop
Rita Tamayo
Abhyudai Singh
Aimee Shen
Unique growth and morphology properties of Clade 5 Clostridioides difficile strains revealed by single-cell time-lapse microscopy.
PLoS Pathogens
title Unique growth and morphology properties of Clade 5 Clostridioides difficile strains revealed by single-cell time-lapse microscopy.
title_full Unique growth and morphology properties of Clade 5 Clostridioides difficile strains revealed by single-cell time-lapse microscopy.
title_fullStr Unique growth and morphology properties of Clade 5 Clostridioides difficile strains revealed by single-cell time-lapse microscopy.
title_full_unstemmed Unique growth and morphology properties of Clade 5 Clostridioides difficile strains revealed by single-cell time-lapse microscopy.
title_short Unique growth and morphology properties of Clade 5 Clostridioides difficile strains revealed by single-cell time-lapse microscopy.
title_sort unique growth and morphology properties of clade 5 clostridioides difficile strains revealed by single cell time lapse microscopy
url https://doi.org/10.1371/journal.ppat.1013155
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