Identification of dfrA14 in two distinct plasmids conferring trimethoprim resistance in Actinobacillus pleuropneumoniae

Janine T Bossé; Yanwen Li; Stephanie Walker; Tom Atherton; Roberto Fernandez Crespo; Susanna M Williamson; Jon Rogers; Roy R Chaudhuri; Lucy A Weinert; Olusegun Oshota; Matthew Holden; Duncan J Maskell; Alexander W Tucker; Brendan W Wren; Andrew N Rycroft; Paul R Langford; BRaDP1T Consortium

doi:10.1093/jac/dkv121

Identification of dfrA14 in two distinct plasmids conferring trimethoprim resistance in Actinobacillus pleuropneumoniae

Janine T Bossé, Yanwen Li, Stephanie Walker, Tom Atherton, Roberto Fernandez Crespo, Susanna M Williamson, Jon Rogers, Roy R Chaudhuri, Lucy A Weinert, Olusegun Oshota, Matthew Holden, Duncan J Maskell, Alexander W Tucker, Brendan W Wren, Andrew N Rycroft, Paul R Langford, BRaDP1T Consortium

Research output: Contribution to journal › Article › peer-review

Abstract

Objectives: The objective of this study was to determine the distribution and genetic basis of trimethoprim resistance in Actinobacillus pleuropneumoniae isolates from pigs in England.

Methods: Clinical isolates collected between 1998 and 2011 were tested for resistance to trimethoprim and sulphonamide. The genetic basis of trimethoprim resistance was determined by shotgun WGS analysis and the subsequent isolation and sequencing of plasmids.

Results: A total of 16 (out of 106) A. pleuropneumoniae isolates were resistant to both trimethoprim (MIC > 32 mg/L) and sulfisoxazole (MIC ≥ 256 mg/L), and a further 32 were resistant only to sulfisoxazole (MIC ≥256 mg/L). Genome
sequence data for the trimethoprim-resistant isolates revealed the presence of the dfrA14 dihydrofolate reductase gene. The distribution of plasmid sequences in multiple contigs suggested the presence of two distinct dfrA14-containing
plasmids in different isolates, which was confirmed by plasmid isolation and sequencing. Both plasmids encoded mobilization genes, the sulphonamide resistance gene sul2, as well as dfrA14 inserted into strA, a streptomycin-resistance-associated gene, although the gene order differed between the two plasmids. One of the plasmids further encoded the strB streptomycin-resistance-associated gene.

Conclusions: This is the first description of mobilizable plasmids conferring trimethoprim resistance in A. pleuropneumoniae and, to our knowledge, the first report of dfrA14 in any member of the Pasteurellaceae. The identification of dfrA14 conferring trimethoprim resistance in A. pleuropneumoniae isolates will facilitate PCR screens for resistance to this important antimicrobial.

Original language	English
Pages (from-to)	2217-2222
Number of pages	6
Journal	Journal of Antimicrobial Chemotherapy
Volume	70
Issue number	8
Early online date	8 May 2015
DOIs	https://doi.org/10.1093/jac/dkv121
Publication status	Published - Aug 2015

Keywords

Animal infections
Antibiotic resistance
Respiratory tract

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10.1093/jac/dkv121

J. Antimicrob. Chemother.-2015-Bossé-2217-22
# The Author 2015. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/ 4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
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Bossé, J. T., Li, Y., Walker, S., Atherton, T., Fernandez Crespo, R., Williamson, S. M., Rogers, J., Chaudhuri, R. R., Weinert, L. A., Oshota, O., Holden, M., Maskell, D. J., Tucker, A. W., Wren, B. W., Rycroft, A. N., Langford, P. R., & BRaDP1T Consortium (2015). Identification of dfrA14 in two distinct plasmids conferring trimethoprim resistance in Actinobacillus pleuropneumoniae. Journal of Antimicrobial Chemotherapy, 70(8), 2217-2222. https://doi.org/10.1093/jac/dkv121

@article{69895b2a5a404a5c93f7e8d0cf179ee2,

title = "Identification of dfrA14 in two distinct plasmids conferring trimethoprim resistance in Actinobacillus pleuropneumoniae",

abstract = "Objectives: The objective of this study was to determine the distribution and genetic basis of trimethoprim resistance in Actinobacillus pleuropneumoniae isolates from pigs in England. Methods: Clinical isolates collected between 1998 and 2011 were tested for resistance to trimethoprim and sulphonamide. The genetic basis of trimethoprim resistance was determined by shotgun WGS analysis and the subsequent isolation and sequencing of plasmids. Results: A total of 16 (out of 106) A. pleuropneumoniae isolates were resistant to both trimethoprim (MIC > 32 mg/L) and sulfisoxazole (MIC ≥ 256 mg/L), and a further 32 were resistant only to sulfisoxazole (MIC ≥256 mg/L). Genome sequence data for the trimethoprim-resistant isolates revealed the presence of the dfrA14 dihydrofolate reductase gene. The distribution of plasmid sequences in multiple contigs suggested the presence of two distinct dfrA14-containing plasmids in different isolates, which was confirmed by plasmid isolation and sequencing. Both plasmids encoded mobilization genes, the sulphonamide resistance gene sul2, as well as dfrA14 inserted into strA, a streptomycin-resistance-associated gene, although the gene order differed between the two plasmids. One of the plasmids further encoded the strB streptomycin-resistance-associated gene. Conclusions: This is the first description of mobilizable plasmids conferring trimethoprim resistance in A. pleuropneumoniae and, to our knowledge, the first report of dfrA14 in any member of the Pasteurellaceae. The identification of dfrA14 conferring trimethoprim resistance in A. pleuropneumoniae isolates will facilitate PCR screens for resistance to this important antimicrobial.",

keywords = "Animal infections, Antibiotic resistance, Respiratory tract",

author = "Boss{\'e}, {Janine T} and Yanwen Li and Stephanie Walker and Tom Atherton and {Fernandez Crespo}, Roberto and Williamson, {Susanna M} and Jon Rogers and Chaudhuri, {Roy R} and Weinert, {Lucy A} and Olusegun Oshota and Matthew Holden and Maskell, {Duncan J} and Tucker, {Alexander W} and Wren, {Brendan W} and Rycroft, {Andrew N} and Langford, {Paul R} and {BRaDP1T Consortium}",

note = "This work was supported by a Longer and Larger (LoLa) grant from the Biotechnology and Biological Sciences Research Council (grant numbers BB/G020744/1, BB/G019177/1, BB/G019274/1 and BB/G018553/1), the UK Department for Environment, Food and Rural Affairs, and Zoetis(formerly Pfizer Animal Health) awarded to the Bacterial Respiratory Diseases of Pigs-1 Technology (BRaDP1T) Consortium. M. T. G. H. was supported by the Wellcome Trust (grant number 098051). The MIC work was funded from the former AHVLA{\textquoteright}s Research and Development Internal Investment Fund (grant number RD0030c). ",

year = "2015",

month = aug,

doi = "10.1093/jac/dkv121",

language = "English",

volume = "70",

pages = "2217--2222",

journal = "Journal of Antimicrobial Chemotherapy",

issn = "0305-7453",

publisher = "Oxford University Press",

number = "8",

}

Bossé, JT, Li, Y, Walker, S, Atherton, T, Fernandez Crespo, R, Williamson, SM, Rogers, J, Chaudhuri, RR, Weinert, LA, Oshota, O, Holden, M, Maskell, DJ, Tucker, AW, Wren, BW, Rycroft, AN, Langford, PR & BRaDP1T Consortium 2015, 'Identification of dfrA14 in two distinct plasmids conferring trimethoprim resistance in Actinobacillus pleuropneumoniae', Journal of Antimicrobial Chemotherapy, vol. 70, no. 8, pp. 2217-2222. https://doi.org/10.1093/jac/dkv121

TY - JOUR

T1 - Identification of dfrA14 in two distinct plasmids conferring trimethoprim resistance in Actinobacillus pleuropneumoniae

AU - Bossé, Janine T

AU - Li, Yanwen

AU - Walker, Stephanie

AU - Atherton, Tom

AU - Fernandez Crespo, Roberto

AU - Williamson, Susanna M

AU - Rogers, Jon

AU - Chaudhuri, Roy R

AU - Weinert, Lucy A

AU - Oshota, Olusegun

AU - Holden, Matthew

AU - Maskell, Duncan J

AU - Tucker, Alexander W

AU - Wren, Brendan W

AU - Rycroft, Andrew N

AU - Langford, Paul R

AU - BRaDP1T Consortium

N1 - This work was supported by a Longer and Larger (LoLa) grant from the Biotechnology and Biological Sciences Research Council (grant numbers BB/G020744/1, BB/G019177/1, BB/G019274/1 and BB/G018553/1), the UK Department for Environment, Food and Rural Affairs, and Zoetis(formerly Pfizer Animal Health) awarded to the Bacterial Respiratory Diseases of Pigs-1 Technology (BRaDP1T) Consortium. M. T. G. H. was supported by the Wellcome Trust (grant number 098051). The MIC work was funded from the former AHVLA’s Research and Development Internal Investment Fund (grant number RD0030c).

PY - 2015/8

Y1 - 2015/8

N2 - Objectives: The objective of this study was to determine the distribution and genetic basis of trimethoprim resistance in Actinobacillus pleuropneumoniae isolates from pigs in England. Methods: Clinical isolates collected between 1998 and 2011 were tested for resistance to trimethoprim and sulphonamide. The genetic basis of trimethoprim resistance was determined by shotgun WGS analysis and the subsequent isolation and sequencing of plasmids. Results: A total of 16 (out of 106) A. pleuropneumoniae isolates were resistant to both trimethoprim (MIC > 32 mg/L) and sulfisoxazole (MIC ≥ 256 mg/L), and a further 32 were resistant only to sulfisoxazole (MIC ≥256 mg/L). Genome sequence data for the trimethoprim-resistant isolates revealed the presence of the dfrA14 dihydrofolate reductase gene. The distribution of plasmid sequences in multiple contigs suggested the presence of two distinct dfrA14-containing plasmids in different isolates, which was confirmed by plasmid isolation and sequencing. Both plasmids encoded mobilization genes, the sulphonamide resistance gene sul2, as well as dfrA14 inserted into strA, a streptomycin-resistance-associated gene, although the gene order differed between the two plasmids. One of the plasmids further encoded the strB streptomycin-resistance-associated gene. Conclusions: This is the first description of mobilizable plasmids conferring trimethoprim resistance in A. pleuropneumoniae and, to our knowledge, the first report of dfrA14 in any member of the Pasteurellaceae. The identification of dfrA14 conferring trimethoprim resistance in A. pleuropneumoniae isolates will facilitate PCR screens for resistance to this important antimicrobial.

AB - Objectives: The objective of this study was to determine the distribution and genetic basis of trimethoprim resistance in Actinobacillus pleuropneumoniae isolates from pigs in England. Methods: Clinical isolates collected between 1998 and 2011 were tested for resistance to trimethoprim and sulphonamide. The genetic basis of trimethoprim resistance was determined by shotgun WGS analysis and the subsequent isolation and sequencing of plasmids. Results: A total of 16 (out of 106) A. pleuropneumoniae isolates were resistant to both trimethoprim (MIC > 32 mg/L) and sulfisoxazole (MIC ≥ 256 mg/L), and a further 32 were resistant only to sulfisoxazole (MIC ≥256 mg/L). Genome sequence data for the trimethoprim-resistant isolates revealed the presence of the dfrA14 dihydrofolate reductase gene. The distribution of plasmid sequences in multiple contigs suggested the presence of two distinct dfrA14-containing plasmids in different isolates, which was confirmed by plasmid isolation and sequencing. Both plasmids encoded mobilization genes, the sulphonamide resistance gene sul2, as well as dfrA14 inserted into strA, a streptomycin-resistance-associated gene, although the gene order differed between the two plasmids. One of the plasmids further encoded the strB streptomycin-resistance-associated gene. Conclusions: This is the first description of mobilizable plasmids conferring trimethoprim resistance in A. pleuropneumoniae and, to our knowledge, the first report of dfrA14 in any member of the Pasteurellaceae. The identification of dfrA14 conferring trimethoprim resistance in A. pleuropneumoniae isolates will facilitate PCR screens for resistance to this important antimicrobial.

KW - Animal infections

KW - Antibiotic resistance

KW - Respiratory tract

U2 - 10.1093/jac/dkv121

DO - 10.1093/jac/dkv121

M3 - Article

C2 - 25957382

SN - 0305-7453

VL - 70

SP - 2217

EP - 2222

JO - Journal of Antimicrobial Chemotherapy

JF - Journal of Antimicrobial Chemotherapy

IS - 8

ER -

Identification of dfrA14 in two distinct plasmids conferring trimethoprim resistance in Actinobacillus pleuropneumoniae

Abstract

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