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Curing vector for IncI1 plasmids and its use to provide evidence for a metabolic burden of IncI1 CTX-M-1 plasmid pIFM3791 on Klebsiella pneumoniae

Freire Martín, I, Thomas, CM, Laing, E, AbuOun, M, La Ragione, RM and Woodward, MJ (2016) Curing vector for IncI1 plasmids and its use to provide evidence for a metabolic burden of IncI1 CTX-M-1 plasmid pIFM3791 on Klebsiella pneumoniae Journal of Medical Microbiology, 65. pp. 611-618.

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Abstract

Using a sequence based approach we previously identified an IncI1 CTX-M-1 plasmid, pIFM3791, on a single pig farm in the UK that was harboured by K. pneumoniae, Escherichia coli and Salmonella enterica serotype 4,5,12,i:-. To test the hypothesis that the plasmid had spread rapidly into these differing host bacteria we wished to assess whether the plasmid conferred a fitness advantage. To do this an IncI1 curing vector was constructed and used to displace the IncI1 CTX-M-1 plasmids from K. pneumoniae strain B3791 and several other unrelated IncI1 harbouring strains indicating the potential wider application of the curing plasmid. The IncI1 CTX-M-1 plasmid was re-introduced by conjugation into the cured K. pneumoniae strain and also a naturally IncI1 plasmid free S. enterica serotype 4,5,12,i:-, S348/1. Original, cured and complemented strains were tested for metabolic competence using BiologTM technology and in competitive growth, association to mammalian cells and biofilm formation experiments. The plasmid-cured K. pneumoniae strain grew more rapidly than either the original plasmid-carrying strain or plasmid-complemented strains in competition experiments. Additionally, the plasmid-cured strain was significantly better at respiring with L-sorbose as a carbon source and putrescine, γ-amino-n-butyric acid, L-alanine, L-proline as a nitrogen sources. By contrast, no differences in phenotype were found when comparing plasmid harbouring and plasmid free S. enterica S348/11. In conclusion, the IncI1 curing vector successfully displaced multiple IncI plasmids. The IncI1 CTX-M1 plasmid conferred a growth disadvantage upon K. pneumoniae, possibly by imposing a metabolic burden the mechanism of which remains to be determined.

Item Type: Article
Subjects : Veterinary Medicine
Divisions : Faculty of Health and Medical Sciences > School of Veterinary Medicine
Authors :
AuthorsEmailORCID
Freire Martín, IUNSPECIFIEDUNSPECIFIED
Thomas, CMUNSPECIFIEDUNSPECIFIED
Laing, EUNSPECIFIEDUNSPECIFIED
AbuOun, MUNSPECIFIEDUNSPECIFIED
La Ragione, RMUNSPECIFIEDUNSPECIFIED
Woodward, MJUNSPECIFIEDUNSPECIFIED
Date : July 2016
Identification Number : 10.1099/jmm.0.000271
Copyright Disclaimer : This is the accepted version of the following article: Curing vector for IncI1 plasmids and its use to provide evidence for a metabolic burden of IncI1 CTX-M-1 plasmid pIFM3791 on Klebsiella pneumonia. The published version is available at https://dx.doi.org/10.1099/jmm.0.000271
Depositing User : Symplectic Elements
Date Deposited : 13 Jul 2016 10:00
Last Modified : 29 Nov 2016 15:26
URI: http://epubs.surrey.ac.uk/id/eprint/811198

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