Necrotic Enteritis-Derived <named-content content-type="genus-species">Clostridium perfringens</named-content> Strain with Three Closely Related Independently Conjugative Toxin and Antibiotic Resistance Plasmids

ABSTRACT The pathogenesis of avian necrotic enteritis involves NetB, a pore-forming toxin produced by virulent avian isolates of Clostridium perfringens type A. To determine the location and mobility of the netB structural gene, we examined a derivative of the tetracycline-resistant necrotic enterit...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Trudi L. Bannam, Xu-Xia Yan, Paul F. Harrison, Torsten Seemann, Anthony L. Keyburn, Christopher Stubenrauch, Lakmini H. Weeramantri, Jackie K. Cheung, Bruce A. McClane, John D. Boyce, Robert J. Moore, Julian I. Rood
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2011
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/68ad830b441c41ef88e3be0903131543
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:68ad830b441c41ef88e3be0903131543
record_format	dspace
spelling	oai:doaj.org-article:68ad830b441c41ef88e3be09031315432021-11-15T15:38:57ZNecrotic Enteritis-Derived <named-content content-type="genus-species">Clostridium perfringens</named-content> Strain with Three Closely Related Independently Conjugative Toxin and Antibiotic Resistance Plasmids10.1128/mBio.00190-112150-7511https://doaj.org/article/68ad830b441c41ef88e3be09031315432011-11-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00190-11https://doaj.org/toc/2150-7511ABSTRACT The pathogenesis of avian necrotic enteritis involves NetB, a pore-forming toxin produced by virulent avian isolates of Clostridium perfringens type A. To determine the location and mobility of the netB structural gene, we examined a derivative of the tetracycline-resistant necrotic enteritis strain EHE-NE18, in which netB was insertionally inactivated by the chloramphenicol and thiamphenicol resistance gene catP. Both tetracycline and thiamphenicol resistance could be transferred either together or separately to a recipient strain in plate matings. The separate transconjugants could act as donors in subsequent matings, which demonstrated that the tetracycline resistance determinant and the netB gene were present on different conjugative elements. Large plasmids were isolated from the transconjugants and analyzed by high-throughput sequencing. Analysis of the resultant data indicated that there were actually three large conjugative plasmids present in the original strain, each with its own toxin or antibiotic resistance locus. Each plasmid contained a highly conserved 40-kb region that included plasmid replication and transfer regions that were closely related to the 47-kb conjugative tetracycline resistance plasmid pCW3 from C. perfringens. The plasmids were as follows: (i) a conjugative 49-kb tetracycline resistance plasmid that was very similar to pCW3, (ii) a conjugative 82-kb plasmid that contained the netB gene and other potential virulence genes, and (iii) a 70-kb plasmid that carried the cpb2 gene, which encodes a different pore-forming toxin, beta2 toxin. IMPORTANCE The anaerobic bacterium Clostridium perfringens can cause an avian gastrointestinal disease known as necrotic enteritis. Disease pathogenesis is not well understood, although the plasmid-encoded pore-forming toxin NetB, is an important virulence factor. In this work, we have shown that the plasmid that carries the netB gene is conjugative and has a 40-kb region that is very similar to replication and transfer regions found within each of the sequenced conjugative plasmids from C. perfringens. We also showed that this strain contained two additional large plasmids that were also conjugative and carried a similar 40-kb region. One of these plasmids encoded beta2 toxin, and the other encoded tetracycline resistance. To our knowledge, this is the first report of a bacterial strain that carries three closely related but different independently conjugative plasmids. These results have significant implications for our understanding of the transmission of virulence and antibiotic resistance genes in pathogenic bacteria.Trudi L. BannamXu-Xia YanPaul F. HarrisonTorsten SeemannAnthony L. KeyburnChristopher StubenrauchLakmini H. WeeramantriJackie K. CheungBruce A. McClaneJohn D. BoyceRobert J. MooreJulian I. RoodAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 2, Iss 5 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Trudi L. Bannam Xu-Xia Yan Paul F. Harrison Torsten Seemann Anthony L. Keyburn Christopher Stubenrauch Lakmini H. Weeramantri Jackie K. Cheung Bruce A. McClane John D. Boyce Robert J. Moore Julian I. Rood Necrotic Enteritis-Derived <named-content content-type="genus-species">Clostridium perfringens</named-content> Strain with Three Closely Related Independently Conjugative Toxin and Antibiotic Resistance Plasmids
description	ABSTRACT The pathogenesis of avian necrotic enteritis involves NetB, a pore-forming toxin produced by virulent avian isolates of Clostridium perfringens type A. To determine the location and mobility of the netB structural gene, we examined a derivative of the tetracycline-resistant necrotic enteritis strain EHE-NE18, in which netB was insertionally inactivated by the chloramphenicol and thiamphenicol resistance gene catP. Both tetracycline and thiamphenicol resistance could be transferred either together or separately to a recipient strain in plate matings. The separate transconjugants could act as donors in subsequent matings, which demonstrated that the tetracycline resistance determinant and the netB gene were present on different conjugative elements. Large plasmids were isolated from the transconjugants and analyzed by high-throughput sequencing. Analysis of the resultant data indicated that there were actually three large conjugative plasmids present in the original strain, each with its own toxin or antibiotic resistance locus. Each plasmid contained a highly conserved 40-kb region that included plasmid replication and transfer regions that were closely related to the 47-kb conjugative tetracycline resistance plasmid pCW3 from C. perfringens. The plasmids were as follows: (i) a conjugative 49-kb tetracycline resistance plasmid that was very similar to pCW3, (ii) a conjugative 82-kb plasmid that contained the netB gene and other potential virulence genes, and (iii) a 70-kb plasmid that carried the cpb2 gene, which encodes a different pore-forming toxin, beta2 toxin. IMPORTANCE The anaerobic bacterium Clostridium perfringens can cause an avian gastrointestinal disease known as necrotic enteritis. Disease pathogenesis is not well understood, although the plasmid-encoded pore-forming toxin NetB, is an important virulence factor. In this work, we have shown that the plasmid that carries the netB gene is conjugative and has a 40-kb region that is very similar to replication and transfer regions found within each of the sequenced conjugative plasmids from C. perfringens. We also showed that this strain contained two additional large plasmids that were also conjugative and carried a similar 40-kb region. One of these plasmids encoded beta2 toxin, and the other encoded tetracycline resistance. To our knowledge, this is the first report of a bacterial strain that carries three closely related but different independently conjugative plasmids. These results have significant implications for our understanding of the transmission of virulence and antibiotic resistance genes in pathogenic bacteria.
format	article
author	Trudi L. Bannam Xu-Xia Yan Paul F. Harrison Torsten Seemann Anthony L. Keyburn Christopher Stubenrauch Lakmini H. Weeramantri Jackie K. Cheung Bruce A. McClane John D. Boyce Robert J. Moore Julian I. Rood
author_facet	Trudi L. Bannam Xu-Xia Yan Paul F. Harrison Torsten Seemann Anthony L. Keyburn Christopher Stubenrauch Lakmini H. Weeramantri Jackie K. Cheung Bruce A. McClane John D. Boyce Robert J. Moore Julian I. Rood
author_sort	Trudi L. Bannam
title	Necrotic Enteritis-Derived <named-content content-type="genus-species">Clostridium perfringens</named-content> Strain with Three Closely Related Independently Conjugative Toxin and Antibiotic Resistance Plasmids
title_short	Necrotic Enteritis-Derived <named-content content-type="genus-species">Clostridium perfringens</named-content> Strain with Three Closely Related Independently Conjugative Toxin and Antibiotic Resistance Plasmids
title_full	Necrotic Enteritis-Derived <named-content content-type="genus-species">Clostridium perfringens</named-content> Strain with Three Closely Related Independently Conjugative Toxin and Antibiotic Resistance Plasmids
title_fullStr	Necrotic Enteritis-Derived <named-content content-type="genus-species">Clostridium perfringens</named-content> Strain with Three Closely Related Independently Conjugative Toxin and Antibiotic Resistance Plasmids
title_full_unstemmed	Necrotic Enteritis-Derived <named-content content-type="genus-species">Clostridium perfringens</named-content> Strain with Three Closely Related Independently Conjugative Toxin and Antibiotic Resistance Plasmids
title_sort	necrotic enteritis-derived <named-content content-type="genus-species">clostridium perfringens</named-content> strain with three closely related independently conjugative toxin and antibiotic resistance plasmids
publisher	American Society for Microbiology
publishDate	2011
url	https://doaj.org/article/68ad830b441c41ef88e3be0903131543
work_keys_str_mv	AT trudilbannam necroticenteritisderivednamedcontentcontenttypegenusspeciesclostridiumperfringensnamedcontentstrainwiththreecloselyrelatedindependentlyconjugativetoxinandantibioticresistanceplasmids AT xuxiayan necroticenteritisderivednamedcontentcontenttypegenusspeciesclostridiumperfringensnamedcontentstrainwiththreecloselyrelatedindependentlyconjugativetoxinandantibioticresistanceplasmids AT paulfharrison necroticenteritisderivednamedcontentcontenttypegenusspeciesclostridiumperfringensnamedcontentstrainwiththreecloselyrelatedindependentlyconjugativetoxinandantibioticresistanceplasmids AT torstenseemann necroticenteritisderivednamedcontentcontenttypegenusspeciesclostridiumperfringensnamedcontentstrainwiththreecloselyrelatedindependentlyconjugativetoxinandantibioticresistanceplasmids AT anthonylkeyburn necroticenteritisderivednamedcontentcontenttypegenusspeciesclostridiumperfringensnamedcontentstrainwiththreecloselyrelatedindependentlyconjugativetoxinandantibioticresistanceplasmids AT christopherstubenrauch necroticenteritisderivednamedcontentcontenttypegenusspeciesclostridiumperfringensnamedcontentstrainwiththreecloselyrelatedindependentlyconjugativetoxinandantibioticresistanceplasmids AT lakminihweeramantri necroticenteritisderivednamedcontentcontenttypegenusspeciesclostridiumperfringensnamedcontentstrainwiththreecloselyrelatedindependentlyconjugativetoxinandantibioticresistanceplasmids AT jackiekcheung necroticenteritisderivednamedcontentcontenttypegenusspeciesclostridiumperfringensnamedcontentstrainwiththreecloselyrelatedindependentlyconjugativetoxinandantibioticresistanceplasmids AT bruceamcclane necroticenteritisderivednamedcontentcontenttypegenusspeciesclostridiumperfringensnamedcontentstrainwiththreecloselyrelatedindependentlyconjugativetoxinandantibioticresistanceplasmids AT johndboyce necroticenteritisderivednamedcontentcontenttypegenusspeciesclostridiumperfringensnamedcontentstrainwiththreecloselyrelatedindependentlyconjugativetoxinandantibioticresistanceplasmids AT robertjmoore necroticenteritisderivednamedcontentcontenttypegenusspeciesclostridiumperfringensnamedcontentstrainwiththreecloselyrelatedindependentlyconjugativetoxinandantibioticresistanceplasmids AT julianirood necroticenteritisderivednamedcontentcontenttypegenusspeciesclostridiumperfringensnamedcontentstrainwiththreecloselyrelatedindependentlyconjugativetoxinandantibioticresistanceplasmids
_version_	1718427828885651456

Necrotic Enteritis-Derived <named-content content-type="genus-species">Clostridium perfringens</named-content> Strain with Three Closely Related Independently Conjugative Toxin and Antibiotic Resistance Plasmids

Ejemplares similares