Increased sporulation underpins adaptation of Clostridium difficile strain 630 to a biologically–relevant faecal environment, with implications for pathogenicity

Nigel G Ternan, Nicola Moore, Deborah Smyth, Gordon McDougall, James William Allwood, Susan Verrall, Chris IR Gill, James Dooley, Geoff McMullan

Research output: Contribution to journalArticle

Abstract

Clostridium difficile virulence is driven primarily by the processes of toxinogenesis and sporulation, however many in vitro experimental systems for studying C. difficile physiology have arguably limited relevance to the human colonic environment. We therefore created a more physiologically–relevant model of the colonic milieu to study gut pathogen biology, incorporating human faecal water (FW) into growth media and assessing the physiological effects of this on C. difficile strain 630. We identified a novel set of C. difficile–derived metabolites in culture supernatants, including hexanoyl– and pentanoyl–amino acid derivatives by LC-MSn. Growth of C. difficile strain 630 in FW media resulted in increased cell length without altering growth rate and RNA sequencing identified 889 transcripts as differentially expressed (p<0.001). Significantly, up to 300–fold increases in the expression of sporulation–associated genes were observed in FW media–grown cells, along with reductions in motility and toxin genes’ expression. Moreover, the expression of classical stress–response genes did not change, showing that C. difficile is well–adapted to this faecal milieu. Using our novel approach we have shown that interaction with FW causes fundamental changes in C. difficile biology that will lead to increased disease transmissibility.
LanguageEnglish
Pages1-16
Number of pages16
JournalScientific Reports
Volume8
DOIs
Publication statusPublished - 12 Nov 2018

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Clostridium difficile
Virulence
Water
Growth
RNA Sequence Analysis
Genes
Gene Expression
Amino Acids

Keywords

  • C. difficile, sporulation, motility, virulence, gut health, faecal water, RNAseq, metabolomics

Cite this

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title = "Increased sporulation underpins adaptation of Clostridium difficile strain 630 to a biologically–relevant faecal environment, with implications for pathogenicity",
abstract = "Clostridium difficile virulence is driven primarily by the processes of toxinogenesis and sporulation, however many in vitro experimental systems for studying C. difficile physiology have arguably limited relevance to the human colonic environment. We therefore created a more physiologically–relevant model of the colonic milieu to study gut pathogen biology, incorporating human faecal water (FW) into growth media and assessing the physiological effects of this on C. difficile strain 630. We identified a novel set of C. difficile–derived metabolites in culture supernatants, including hexanoyl– and pentanoyl–amino acid derivatives by LC-MSn. Growth of C. difficile strain 630 in FW media resulted in increased cell length without altering growth rate and RNA sequencing identified 889 transcripts as differentially expressed (p<0.001). Significantly, up to 300–fold increases in the expression of sporulation–associated genes were observed in FW media–grown cells, along with reductions in motility and toxin genes’ expression. Moreover, the expression of classical stress–response genes did not change, showing that C. difficile is well–adapted to this faecal milieu. Using our novel approach we have shown that interaction with FW causes fundamental changes in C. difficile biology that will lead to increased disease transmissibility.",
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Increased sporulation underpins adaptation of Clostridium difficile strain 630 to a biologically–relevant faecal environment, with implications for pathogenicity. / Ternan, Nigel G; Moore, Nicola ; Smyth, Deborah; McDougall, Gordon; Allwood, James William; Verrall, Susan ; Gill, Chris IR; Dooley, James; McMullan, Geoff.

In: Scientific Reports, Vol. 8, 12.11.2018, p. 1-16.

Research output: Contribution to journalArticle

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AU - Ternan, Nigel G

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AU - Smyth, Deborah

AU - McDougall, Gordon

AU - Allwood, James William

AU - Verrall, Susan

AU - Gill, Chris IR

AU - Dooley, James

AU - McMullan, Geoff

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