Isolation and characterisation of four novel bacteriophages infecting clinically relevant PCR ribotypes of Clostridium difficile

Clostridium difficile infection is the leading cause of antibiotic-associated diarrhoea in North America and Europe. The disease is most commonly treated with only a handful of antibiotics, amongst those vancomycin, a so-called last resort therapy. Growing concerns over antibiotic resistance have le...

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Main Author: Whittle, Michaella
Format: Thesis (University of Nottingham only)
Language:English
Published: 2018
Subjects:
Online Access:https://eprints.nottingham.ac.uk/52208/
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author Whittle, Michaella
author_facet Whittle, Michaella
author_sort Whittle, Michaella
building Nottingham Research Data Repository
collection Online Access
description Clostridium difficile infection is the leading cause of antibiotic-associated diarrhoea in North America and Europe. The disease is most commonly treated with only a handful of antibiotics, amongst those vancomycin, a so-called last resort therapy. Growing concerns over antibiotic resistance have led to the desire for more targeted therapies. In this study, we report the isolation and characterisation of four novel bacteriophages, ΦCD08011, ΦCD2301, ΦCD418 and ΦCD1801, able to infect a range of clinically relevant C. difficile PCR ribotypes. The four phages belong to the Myoviridae family of tailed phages, established through the use of transmission electron microscopy and the identification of key genes within their genomes. Host range analysis of the four phages showed that phage ΦCD1801 had the broadest host range, able to infect and lyse all but one of the PCR ribotype 078 isolates tested. Further growth rate characterisation, rate of attachment and determination of burst size were also completed. Finally, SlpA, the main component of C. difficile’s S-layer, was confirmed as the bacterial receptor for this phage. The presence of slpA from the S-layer cassette of PCR ribotype 078 C. difficile strains conferred ΦCD1801 binding to an otherwise resistant strain. These findings will be instrumental in the ability to expand C. difficile phage host range to allow more targeted phages for the treatment of C. difficile infections.
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spelling nottingham-522082025-02-28T14:09:24Z https://eprints.nottingham.ac.uk/52208/ Isolation and characterisation of four novel bacteriophages infecting clinically relevant PCR ribotypes of Clostridium difficile Whittle, Michaella Clostridium difficile infection is the leading cause of antibiotic-associated diarrhoea in North America and Europe. The disease is most commonly treated with only a handful of antibiotics, amongst those vancomycin, a so-called last resort therapy. Growing concerns over antibiotic resistance have led to the desire for more targeted therapies. In this study, we report the isolation and characterisation of four novel bacteriophages, ΦCD08011, ΦCD2301, ΦCD418 and ΦCD1801, able to infect a range of clinically relevant C. difficile PCR ribotypes. The four phages belong to the Myoviridae family of tailed phages, established through the use of transmission electron microscopy and the identification of key genes within their genomes. Host range analysis of the four phages showed that phage ΦCD1801 had the broadest host range, able to infect and lyse all but one of the PCR ribotype 078 isolates tested. Further growth rate characterisation, rate of attachment and determination of burst size were also completed. Finally, SlpA, the main component of C. difficile’s S-layer, was confirmed as the bacterial receptor for this phage. The presence of slpA from the S-layer cassette of PCR ribotype 078 C. difficile strains conferred ΦCD1801 binding to an otherwise resistant strain. These findings will be instrumental in the ability to expand C. difficile phage host range to allow more targeted phages for the treatment of C. difficile infections. 2018-07-12 Thesis (University of Nottingham only) NonPeerReviewed application/pdf en arr https://eprints.nottingham.ac.uk/52208/1/MW_corrected_thesis_Mar18.pdf Whittle, Michaella (2018) Isolation and characterisation of four novel bacteriophages infecting clinically relevant PCR ribotypes of Clostridium difficile. PhD thesis, University of Nottingham. C. difficile bacteriophages genome sequencing alternative therapies
spellingShingle C. difficile
bacteriophages
genome sequencing
alternative therapies
Whittle, Michaella
Isolation and characterisation of four novel bacteriophages infecting clinically relevant PCR ribotypes of Clostridium difficile
title Isolation and characterisation of four novel bacteriophages infecting clinically relevant PCR ribotypes of Clostridium difficile
title_full Isolation and characterisation of four novel bacteriophages infecting clinically relevant PCR ribotypes of Clostridium difficile
title_fullStr Isolation and characterisation of four novel bacteriophages infecting clinically relevant PCR ribotypes of Clostridium difficile
title_full_unstemmed Isolation and characterisation of four novel bacteriophages infecting clinically relevant PCR ribotypes of Clostridium difficile
title_short Isolation and characterisation of four novel bacteriophages infecting clinically relevant PCR ribotypes of Clostridium difficile
title_sort isolation and characterisation of four novel bacteriophages infecting clinically relevant pcr ribotypes of clostridium difficile
topic C. difficile
bacteriophages
genome sequencing
alternative therapies
url https://eprints.nottingham.ac.uk/52208/