Characterization of pFOX-7a, a conjugative IncL/M plasmid encoding the FOX-7 AmpC-type β-lactamase, involved in a large outbreak in a neonatal intensive care unit

Vincenzo Di Pilato, Fabio Arena, Tommaso Giani, Viola Conte, Stefania Cresti, Gian M aria Rossolini

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

OBJECTIVES: FOX-type enzymes are a lineage of AmpC-type β-lactamases from Aeromonas spp. whose genes have been mobilized to plasmids spreading among Enterobacteriaceae, where they can be responsible for resistance to extended-spectrum cephalosporins and β-lactamase inhibitor combinations. Little is known about the genetic context and plasmid vehicles of bla(FOX) determinants. Here, we have characterized a plasmid encoding the FOX-7 β-lactamase, which was involved in a large outbreak caused by two Klebsiella pneumoniae clones in a neonatal intensive care unit.

METHODS: Plasmid transferability was tested in conjugation experiments using Escherichia coli recipients. Plasmids from different strains were compared by restriction profiling and PCR mapping. The complete sequence of pFOX-7a plasmid was determined by a next-generation sequencing approach followed by gap filling using PCR and sequencing.

RESULTS: An apparently identical conjugative plasmid encoding FOX-7 was detected in representatives of the K. pneumoniae clones that caused the outbreak and in sporadic FOX-7-producing strains of other species from the same ward. The plasmid, named pFOX-7a, has an IncL/M-type backbone and two separate resistance modules including a Tn3-like transposon and a novel Tn1696 derivative, named Tn6234, which carries an integron platform, a hybrid (but still functional) mercury resistance module and a novel putative transposon of original structure, named Tn6240, associated with the bla(FOX-7) gene.

CONCLUSIONS: pFOX-7a is the first completely characterized plasmid encoding a FOX-type β-lactamase. The bla(FOX-7) gene was associated with a putative transposable element of original structure, which was likely involved in its mobilization from the Aeromonas metagenome.

Original languageEnglish
Pages (from-to)2620-2624
Number of pages5
JournalJournal of Antimicrobial Chemotherapy
Volume69
Issue number10
DOIs
Publication statusPublished - Oct 1 2014

Fingerprint

Neonatal Intensive Care Units
Disease Outbreaks
Plasmids
Aeromonas
Klebsiella pneumoniae
Clone Cells
Metagenome
Integrons
Genes
Polymerase Chain Reaction
DNA Transposable Elements
Enterobacteriaceae
Cephalosporins
Mercury
Escherichia coli
Enzymes

Keywords

  • class C β-lactamases
  • FOX-type β-lactamase
  • Gram-negative
  • transposons

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Characterization of pFOX-7a, a conjugative IncL/M plasmid encoding the FOX-7 AmpC-type β-lactamase, involved in a large outbreak in a neonatal intensive care unit. / Di Pilato, Vincenzo; Arena, Fabio; Giani, Tommaso; Conte, Viola; Cresti, Stefania; Rossolini, Gian M aria.

In: Journal of Antimicrobial Chemotherapy, Vol. 69, No. 10, 01.10.2014, p. 2620-2624.

Research output: Contribution to journalArticle

Di Pilato, Vincenzo ; Arena, Fabio ; Giani, Tommaso ; Conte, Viola ; Cresti, Stefania ; Rossolini, Gian M aria. / Characterization of pFOX-7a, a conjugative IncL/M plasmid encoding the FOX-7 AmpC-type β-lactamase, involved in a large outbreak in a neonatal intensive care unit. In: Journal of Antimicrobial Chemotherapy. 2014 ; Vol. 69, No. 10. pp. 2620-2624.
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abstract = "OBJECTIVES: FOX-type enzymes are a lineage of AmpC-type β-lactamases from Aeromonas spp. whose genes have been mobilized to plasmids spreading among Enterobacteriaceae, where they can be responsible for resistance to extended-spectrum cephalosporins and β-lactamase inhibitor combinations. Little is known about the genetic context and plasmid vehicles of bla(FOX) determinants. Here, we have characterized a plasmid encoding the FOX-7 β-lactamase, which was involved in a large outbreak caused by two Klebsiella pneumoniae clones in a neonatal intensive care unit.METHODS: Plasmid transferability was tested in conjugation experiments using Escherichia coli recipients. Plasmids from different strains were compared by restriction profiling and PCR mapping. The complete sequence of pFOX-7a plasmid was determined by a next-generation sequencing approach followed by gap filling using PCR and sequencing.RESULTS: An apparently identical conjugative plasmid encoding FOX-7 was detected in representatives of the K. pneumoniae clones that caused the outbreak and in sporadic FOX-7-producing strains of other species from the same ward. The plasmid, named pFOX-7a, has an IncL/M-type backbone and two separate resistance modules including a Tn3-like transposon and a novel Tn1696 derivative, named Tn6234, which carries an integron platform, a hybrid (but still functional) mercury resistance module and a novel putative transposon of original structure, named Tn6240, associated with the bla(FOX-7) gene.CONCLUSIONS: pFOX-7a is the first completely characterized plasmid encoding a FOX-type β-lactamase. The bla(FOX-7) gene was associated with a putative transposable element of original structure, which was likely involved in its mobilization from the Aeromonas metagenome.",
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AU - Di Pilato, Vincenzo

AU - Arena, Fabio

AU - Giani, Tommaso

AU - Conte, Viola

AU - Cresti, Stefania

AU - Rossolini, Gian M aria

PY - 2014/10/1

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N2 - OBJECTIVES: FOX-type enzymes are a lineage of AmpC-type β-lactamases from Aeromonas spp. whose genes have been mobilized to plasmids spreading among Enterobacteriaceae, where they can be responsible for resistance to extended-spectrum cephalosporins and β-lactamase inhibitor combinations. Little is known about the genetic context and plasmid vehicles of bla(FOX) determinants. Here, we have characterized a plasmid encoding the FOX-7 β-lactamase, which was involved in a large outbreak caused by two Klebsiella pneumoniae clones in a neonatal intensive care unit.METHODS: Plasmid transferability was tested in conjugation experiments using Escherichia coli recipients. Plasmids from different strains were compared by restriction profiling and PCR mapping. The complete sequence of pFOX-7a plasmid was determined by a next-generation sequencing approach followed by gap filling using PCR and sequencing.RESULTS: An apparently identical conjugative plasmid encoding FOX-7 was detected in representatives of the K. pneumoniae clones that caused the outbreak and in sporadic FOX-7-producing strains of other species from the same ward. The plasmid, named pFOX-7a, has an IncL/M-type backbone and two separate resistance modules including a Tn3-like transposon and a novel Tn1696 derivative, named Tn6234, which carries an integron platform, a hybrid (but still functional) mercury resistance module and a novel putative transposon of original structure, named Tn6240, associated with the bla(FOX-7) gene.CONCLUSIONS: pFOX-7a is the first completely characterized plasmid encoding a FOX-type β-lactamase. The bla(FOX-7) gene was associated with a putative transposable element of original structure, which was likely involved in its mobilization from the Aeromonas metagenome.

AB - OBJECTIVES: FOX-type enzymes are a lineage of AmpC-type β-lactamases from Aeromonas spp. whose genes have been mobilized to plasmids spreading among Enterobacteriaceae, where they can be responsible for resistance to extended-spectrum cephalosporins and β-lactamase inhibitor combinations. Little is known about the genetic context and plasmid vehicles of bla(FOX) determinants. Here, we have characterized a plasmid encoding the FOX-7 β-lactamase, which was involved in a large outbreak caused by two Klebsiella pneumoniae clones in a neonatal intensive care unit.METHODS: Plasmid transferability was tested in conjugation experiments using Escherichia coli recipients. Plasmids from different strains were compared by restriction profiling and PCR mapping. The complete sequence of pFOX-7a plasmid was determined by a next-generation sequencing approach followed by gap filling using PCR and sequencing.RESULTS: An apparently identical conjugative plasmid encoding FOX-7 was detected in representatives of the K. pneumoniae clones that caused the outbreak and in sporadic FOX-7-producing strains of other species from the same ward. The plasmid, named pFOX-7a, has an IncL/M-type backbone and two separate resistance modules including a Tn3-like transposon and a novel Tn1696 derivative, named Tn6234, which carries an integron platform, a hybrid (but still functional) mercury resistance module and a novel putative transposon of original structure, named Tn6240, associated with the bla(FOX-7) gene.CONCLUSIONS: pFOX-7a is the first completely characterized plasmid encoding a FOX-type β-lactamase. The bla(FOX-7) gene was associated with a putative transposable element of original structure, which was likely involved in its mobilization from the Aeromonas metagenome.

KW - class C β-lactamases

KW - FOX-type β-lactamase

KW - Gram-negative

KW - transposons

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