Complete sequences, transferability and fitness cost of epidemic IncX plasmids carrying quinolone resistence genes from E. coli isolates of various origin

Kód výsledku v IS VaVaI

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Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Complete sequences, transferability and fitness cost of epidemic IncX plasmids carrying quinolone resistence genes from E. coli isolates of various origin

Popis výsledku v původním jazyce

ncX plasmids are important vehicles of antibiotic resistance genes. Recently, we identified two epidemic lineages of Incx1 and IncX2 plasmids disseminating qnrS1 gene within the population of Escherichia coli. The goal of this study was to determine the complete nucleotide sequence, transferability and persistence of IncX plasmids carrying qnr genes, their impact on the fitness of host bacteria and the role of temperature, antimicrobials and host background on their transfer. Nine plasmids belonging to IncX1 (n=5), IncX2 (n=3) and IncX3 (n=1), carrying qnr genes and originating from wildlife (6 plasmids), wastewater (2) and a dog (1) were analyzed by Roche 454 and MiSeq Illumina. Frequency of plasmid transfer (FOT) was determined as a ratio of transconjugants per recipient. Growth curves of cells carrying the plasmid versus plasmid-free cells were used to determine the plasmid impact on the host (commensal/pathogenic E. coli, Salmonella spp., Pseudomonas aeruginosa). For the representatives of IncX1 (pHP2) and IncX2 (p194) plasmids, the role of temperatures (25-37°C) and concentrations of ciprofloxacin (0.001-2 ?g/ml) as well as the host background on FOT was tested. Persistence of the plasmids was measured by determining the viable counts on the plate with and without the antibiotic. The sequence analysis identified hotspots for acquiring qnr genes in the plasmid backbone. The FOT between the 9 plasmids varied up to 6-folds, depending on the plasmid and the growth phase. Stress factors (temperature and antibiotics) had uneven effects among the representative plasmids. In the presence of ciprofloxacin the FOT of pHP2 significantly increased unlike p194. Lowered temperature enhanced the FOT in p194, while the FOT of pHP2 raised with the increase of temperature. Both plasmids had approximately one fold lower FOT when transferred from laboratory to pathogenic strain than when transferred from pathogenic to pathogenic strain. Impact on the host fitness was observed among several plasmids. Persistence experiments revealed that the host cells were capable of sustaining the plasmid for the whole tested period of time. The results suggest different coping of the host carrying particular IncX plasmid with each stress condition. The study supports the idea that once the plasmid is acquired by a pathogenic host, its transfer frequency within this population is increased. The plasmid persistence experiment suggest low burden on the host cell, likely as a result of evolutionary adaptation between the host bacteria and the plasmid.

Název v anglickém jazyce

Complete sequences, transferability and fitness cost of epidemic IncX plasmids carrying quinolone resistence genes from E. coli isolates of various origin

Popis výsledku anglicky

ncX plasmids are important vehicles of antibiotic resistance genes. Recently, we identified two epidemic lineages of Incx1 and IncX2 plasmids disseminating qnrS1 gene within the population of Escherichia coli. The goal of this study was to determine the complete nucleotide sequence, transferability and persistence of IncX plasmids carrying qnr genes, their impact on the fitness of host bacteria and the role of temperature, antimicrobials and host background on their transfer. Nine plasmids belonging to IncX1 (n=5), IncX2 (n=3) and IncX3 (n=1), carrying qnr genes and originating from wildlife (6 plasmids), wastewater (2) and a dog (1) were analyzed by Roche 454 and MiSeq Illumina. Frequency of plasmid transfer (FOT) was determined as a ratio of transconjugants per recipient. Growth curves of cells carrying the plasmid versus plasmid-free cells were used to determine the plasmid impact on the host (commensal/pathogenic E. coli, Salmonella spp., Pseudomonas aeruginosa). For the representatives of IncX1 (pHP2) and IncX2 (p194) plasmids, the role of temperatures (25-37°C) and concentrations of ciprofloxacin (0.001-2 ?g/ml) as well as the host background on FOT was tested. Persistence of the plasmids was measured by determining the viable counts on the plate with and without the antibiotic. The sequence analysis identified hotspots for acquiring qnr genes in the plasmid backbone. The FOT between the 9 plasmids varied up to 6-folds, depending on the plasmid and the growth phase. Stress factors (temperature and antibiotics) had uneven effects among the representative plasmids. In the presence of ciprofloxacin the FOT of pHP2 significantly increased unlike p194. Lowered temperature enhanced the FOT in p194, while the FOT of pHP2 raised with the increase of temperature. Both plasmids had approximately one fold lower FOT when transferred from laboratory to pathogenic strain than when transferred from pathogenic to pathogenic strain. Impact on the host fitness was observed among several plasmids. Persistence experiments revealed that the host cells were capable of sustaining the plasmid for the whole tested period of time. The results suggest different coping of the host carrying particular IncX plasmid with each stress condition. The study supports the idea that once the plasmid is acquired by a pathogenic host, its transfer frequency within this population is increased. The plasmid persistence experiment suggest low burden on the host cell, likely as a result of evolutionary adaptation between the host bacteria and the plasmid.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

10606 - Microbiology

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2018

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů