Analysis of the spore germination mechanisms of Clostridium difficile

Clostridium difficile is the leading cause of hospital acquired diarrhoea and a major burden to healthcare services worldwide. Endospore production plays a pivotal role in infection and disease transmission, but in order to cause disease these spores must germinate and return to vegetative cell grow...

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Main Author: Burns, David Alexander
Format: Thesis (University of Nottingham only)
Language:English
Published: 2011
Subjects:
Online Access:https://eprints.nottingham.ac.uk/11852/
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author Burns, David Alexander
author_facet Burns, David Alexander
author_sort Burns, David Alexander
building Nottingham Research Data Repository
collection Online Access
description Clostridium difficile is the leading cause of hospital acquired diarrhoea and a major burden to healthcare services worldwide. Endospore production plays a pivotal role in infection and disease transmission, but in order to cause disease these spores must germinate and return to vegetative cell growth. Therefore, knowledge of spore germination is important and may have direct applications in future disease prevention. Germination has been well studied in Bacillus and in some clostridia, but the mechanisms of C. difficile spore germination remain unclear. Apparent homologues of genes important for germination in other spore formers have been identified in the C. difficile genome and ClosTron technology was used to inactivate homologues of sleC, cspA, cspB and cspC (Clostridium perfringens) and cwlJ, sleB and cwlD (Bacillus subtilis) in both C. difficile 630Δerm and a BI/NAP1/027 isolate (a ‘hypervirulent’ type associated with outbreaks of increased disease severity). Using a combination of several different assays to study these mutants in detail, a number of the identified target genes appear to be essential for germination and outgrowth of C. difficile spores. This is the first report of using reverse genetics to study the germination of C. difficile spores and the first gene characterisation by mutagenesis in a BI/NAP1/027 isolate of C. difficile. Furthermore, this study uncovered evidence of significant variation in the sporulation and germination characteristics of different C. difficile strains, but this variation did not appear to be type-associated.
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spelling nottingham-118522025-02-28T11:16:00Z https://eprints.nottingham.ac.uk/11852/ Analysis of the spore germination mechanisms of Clostridium difficile Burns, David Alexander Clostridium difficile is the leading cause of hospital acquired diarrhoea and a major burden to healthcare services worldwide. Endospore production plays a pivotal role in infection and disease transmission, but in order to cause disease these spores must germinate and return to vegetative cell growth. Therefore, knowledge of spore germination is important and may have direct applications in future disease prevention. Germination has been well studied in Bacillus and in some clostridia, but the mechanisms of C. difficile spore germination remain unclear. Apparent homologues of genes important for germination in other spore formers have been identified in the C. difficile genome and ClosTron technology was used to inactivate homologues of sleC, cspA, cspB and cspC (Clostridium perfringens) and cwlJ, sleB and cwlD (Bacillus subtilis) in both C. difficile 630Δerm and a BI/NAP1/027 isolate (a ‘hypervirulent’ type associated with outbreaks of increased disease severity). Using a combination of several different assays to study these mutants in detail, a number of the identified target genes appear to be essential for germination and outgrowth of C. difficile spores. This is the first report of using reverse genetics to study the germination of C. difficile spores and the first gene characterisation by mutagenesis in a BI/NAP1/027 isolate of C. difficile. Furthermore, this study uncovered evidence of significant variation in the sporulation and germination characteristics of different C. difficile strains, but this variation did not appear to be type-associated. 2011-07-15 Thesis (University of Nottingham only) NonPeerReviewed application/pdf en arr https://eprints.nottingham.ac.uk/11852/1/David_Burns_PhD_Thesis.pdf Burns, David Alexander (2011) Analysis of the spore germination mechanisms of Clostridium difficile. PhD thesis, University of Nottingham. Clostridium difficile spore sporulation germination SleC
spellingShingle Clostridium difficile
spore
sporulation
germination
SleC
Burns, David Alexander
Analysis of the spore germination mechanisms of Clostridium difficile
title Analysis of the spore germination mechanisms of Clostridium difficile
title_full Analysis of the spore germination mechanisms of Clostridium difficile
title_fullStr Analysis of the spore germination mechanisms of Clostridium difficile
title_full_unstemmed Analysis of the spore germination mechanisms of Clostridium difficile
title_short Analysis of the spore germination mechanisms of Clostridium difficile
title_sort analysis of the spore germination mechanisms of clostridium difficile
topic Clostridium difficile
spore
sporulation
germination
SleC
url https://eprints.nottingham.ac.uk/11852/