Impact of Phage CDHS-1 on the Transcription, Physiology and Pathogenicity of a <i>Clostridioides difficile</i> Ribotype 027 Strain, R20291

All known <i>Clostridioides difficile</i> phages encode integrases rendering them potentially able to lyse or lysogenise bacterial strains. Here, we observed the infection of the siphovirus, CDHS-1 on a ribotype 027 strain, R20291 and determined the phage and bacterial gene expression pr...

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Autores principales:	Janet Y. Nale, Thekra Sideeq Al-Tayawi, Shaun Heaphy, Martha R. J. Clokie
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	<i>Clostridioides difficile</i> pathogenicity locus PaLoc <i>Galleria mellonella</i> bacteriophage Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/46311e25dd6b43dba4f6d924a10e025d
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spelling	oai:doaj.org-article:46311e25dd6b43dba4f6d924a10e025d2021-11-25T19:13:53ZImpact of Phage CDHS-1 on the Transcription, Physiology and Pathogenicity of a <i>Clostridioides difficile</i> Ribotype 027 Strain, R2029110.3390/v131122621999-4915https://doaj.org/article/46311e25dd6b43dba4f6d924a10e025d2021-11-01T00:00:00Zhttps://www.mdpi.com/1999-4915/13/11/2262https://doaj.org/toc/1999-4915All known <i>Clostridioides difficile</i> phages encode integrases rendering them potentially able to lyse or lysogenise bacterial strains. Here, we observed the infection of the siphovirus, CDHS-1 on a ribotype 027 strain, R20291 and determined the phage and bacterial gene expression profiles, and impacts of phage infection on bacterial physiology and pathogenicity. Using RNA-seq and RT-qPCR we analysed transcriptomic changes during early, mid-log and late phases of phage replication at an MOI of 10. The phage has a 20 min latent period, takes 80 min to lyse cells and a burst size of ~37. All phage genes are highly expressed during at least one time point. The <i>Cro</i>/C1-transcriptional regulator, ssDNA binding protein and helicase are expressed early, the holin is expressed during the mid-log phase and structural proteins are expressed from mid-log to late phase. Most bacterial genes, particularly the metabolism and toxin production/regulatory genes, were downregulated from early phage replication. Phage-resistant strains and lysogens showed reduced virulence during <i>Galleria mellonella</i> colonization as ascertained by the larval survival and expression of growth (10), reproduction (2) and infection (2) marker genes. These data suggest that phage infection both reduces colonization and negatively impacts bacterial pathogenicity, providing encouraging data to support the development of this phage for therapy to treat <i>C. difficile</i> infection.Janet Y. NaleThekra Sideeq Al-TayawiShaun HeaphyMartha R. J. ClokieMDPI AGarticle<i>Clostridioides difficile</i>pathogenicity locusPaLoc<i>Galleria mellonella</i>bacteriophageMicrobiologyQR1-502ENViruses, Vol 13, Iss 2262, p 2262 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	<i>Clostridioides difficile</i> pathogenicity locus PaLoc <i>Galleria mellonella</i> bacteriophage Microbiology QR1-502
spellingShingle	<i>Clostridioides difficile</i> pathogenicity locus PaLoc <i>Galleria mellonella</i> bacteriophage Microbiology QR1-502 Janet Y. Nale Thekra Sideeq Al-Tayawi Shaun Heaphy Martha R. J. Clokie Impact of Phage CDHS-1 on the Transcription, Physiology and Pathogenicity of a <i>Clostridioides difficile</i> Ribotype 027 Strain, R20291
description	All known <i>Clostridioides difficile</i> phages encode integrases rendering them potentially able to lyse or lysogenise bacterial strains. Here, we observed the infection of the siphovirus, CDHS-1 on a ribotype 027 strain, R20291 and determined the phage and bacterial gene expression profiles, and impacts of phage infection on bacterial physiology and pathogenicity. Using RNA-seq and RT-qPCR we analysed transcriptomic changes during early, mid-log and late phases of phage replication at an MOI of 10. The phage has a 20 min latent period, takes 80 min to lyse cells and a burst size of ~37. All phage genes are highly expressed during at least one time point. The <i>Cro</i>/C1-transcriptional regulator, ssDNA binding protein and helicase are expressed early, the holin is expressed during the mid-log phase and structural proteins are expressed from mid-log to late phase. Most bacterial genes, particularly the metabolism and toxin production/regulatory genes, were downregulated from early phage replication. Phage-resistant strains and lysogens showed reduced virulence during <i>Galleria mellonella</i> colonization as ascertained by the larval survival and expression of growth (10), reproduction (2) and infection (2) marker genes. These data suggest that phage infection both reduces colonization and negatively impacts bacterial pathogenicity, providing encouraging data to support the development of this phage for therapy to treat <i>C. difficile</i> infection.
format	article
author	Janet Y. Nale Thekra Sideeq Al-Tayawi Shaun Heaphy Martha R. J. Clokie
author_facet	Janet Y. Nale Thekra Sideeq Al-Tayawi Shaun Heaphy Martha R. J. Clokie
author_sort	Janet Y. Nale
title	Impact of Phage CDHS-1 on the Transcription, Physiology and Pathogenicity of a <i>Clostridioides difficile</i> Ribotype 027 Strain, R20291
title_short	Impact of Phage CDHS-1 on the Transcription, Physiology and Pathogenicity of a <i>Clostridioides difficile</i> Ribotype 027 Strain, R20291
title_full	Impact of Phage CDHS-1 on the Transcription, Physiology and Pathogenicity of a <i>Clostridioides difficile</i> Ribotype 027 Strain, R20291
title_fullStr	Impact of Phage CDHS-1 on the Transcription, Physiology and Pathogenicity of a <i>Clostridioides difficile</i> Ribotype 027 Strain, R20291
title_full_unstemmed	Impact of Phage CDHS-1 on the Transcription, Physiology and Pathogenicity of a <i>Clostridioides difficile</i> Ribotype 027 Strain, R20291
title_sort	impact of phage cdhs-1 on the transcription, physiology and pathogenicity of a <i>clostridioides difficile</i> ribotype 027 strain, r20291
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/46311e25dd6b43dba4f6d924a10e025d
work_keys_str_mv	AT janetynale impactofphagecdhs1onthetranscriptionphysiologyandpathogenicityofaiclostridioidesdifficileiribotype027strainr20291 AT thekrasideeqaltayawi impactofphagecdhs1onthetranscriptionphysiologyandpathogenicityofaiclostridioidesdifficileiribotype027strainr20291 AT shaunheaphy impactofphagecdhs1onthetranscriptionphysiologyandpathogenicityofaiclostridioidesdifficileiribotype027strainr20291 AT martharjclokie impactofphagecdhs1onthetranscriptionphysiologyandpathogenicityofaiclostridioidesdifficileiribotype027strainr20291
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Impact of Phage CDHS-1 on the Transcription, Physiology and Pathogenicity of a <i>Clostridioides difficile</i> Ribotype 027 Strain, R20291

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