First Report of Plasmid-Mediated Colistin Resistance mcr-1 in Escherichia coli from Birds of Prey in Russia

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F62157124%3A16270%2F19%3A43878179" target="_blank" >RIV/62157124:16270/19:43878179 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

First Report of Plasmid-Mediated Colistin Resistance mcr-1 in Escherichia coli from Birds of Prey in Russia

Popis výsledku v původním jazyce

Colistin is listed by World Health Organization as a critically important antibiotic for human medicine. Since the first report of a plasmid-mediated colistin resistance gene in 2015, mcr-1 has spread globally and has been reported in more than 40 countries. The aim of this study was to isolate and characterize Enterobacteriaceae with reduced susceptibility to colistin from Red and Black Kites (Milvus milvus and Milvus migrans, respectively) and to assess their potential risk as reservoirs and vectors of colistin resistance genes. One hundred sixty eight cloacal samples were collected from Kites in Euroasia (Austria n=6, Belgium n=12, Czech Republic n=93, Germany n=41 and Russia n=16) and cultivated on MacConkey agar with colistin (3.5 mgL?1). Enterobacteriaceae isolates with reduced susceptibility to colistin were tested for mcr-1 to mcr-8 genes by polymerase chain reaction and Sanger sequencing. Matrix-Assisted Laser Desorption/Ionization-Time of Flight mass spectrometry was used to identify selected isolates to species level. Susceptibility to a set of 18 antimicrobials was determined using disk diffusion method while broth dilution method was used to determine minimal inhibitory concentration (MIC) to colistin. Conjugation assay based on filter mating was carried to determine the conjugative property of mcr-1-carrying plasmid. Wholegenome sequencing (WGS) was performed using MiSeq (Illumina) platform to analyse the genetic content of selected isolates and investigate plasmids with mcr-1 gene. Overall, 13 (8%, n=168) Enterobacteriaceae isolates showed reduced susceptibility to colistin (Escherichia coli n=9, Klebsiella pneumoniae n=2 and Enterobacter cancerogenus n=2). The gene mcr-1 was positive in one E. coli isolate originating from a Black Kite sampled at Semenovod, Russia. Apart from resistance to colisitn (&gt; 4mg/L), the isolates showed susceptibility to all tested antibiotics. WGS analysis revealed mcr-1 positive isolate belonged to sequence type 2280. Sequencing and plasmid transfer experiments showed the location of mcr-1 on a conjugative IncI2 plasmid pDR164 (59891 bp, GenBank accession number MK542639). Plasmid pDR164 showed similarity to IncI2 plasmids found in E. coli isolates from swine, poultry, human and sewage in USA, Vietnam, Argentina and China, respectively. Comparison of pDR164 with other plasmids revealed high similarity in the backbone as well as the region carrying mcr. Unlike pDR164, plasmids from Vietnam, China and USA carried insertion sequence elements flanking the corresponding mcr gene. We report the first mcr-1 gene in E. coli from wildlife in Russia on a typical IncI2 plasmid. Considering the global spread of plasmid-encoded colistin resistance, migratory birds may represent a vehicle for dissemination of clinically-important antimicrobial resistance bacteria and genes.

Název v anglickém jazyce

First Report of Plasmid-Mediated Colistin Resistance mcr-1 in Escherichia coli from Birds of Prey in Russia

Popis výsledku anglicky

Colistin is listed by World Health Organization as a critically important antibiotic for human medicine. Since the first report of a plasmid-mediated colistin resistance gene in 2015, mcr-1 has spread globally and has been reported in more than 40 countries. The aim of this study was to isolate and characterize Enterobacteriaceae with reduced susceptibility to colistin from Red and Black Kites (Milvus milvus and Milvus migrans, respectively) and to assess their potential risk as reservoirs and vectors of colistin resistance genes. One hundred sixty eight cloacal samples were collected from Kites in Euroasia (Austria n=6, Belgium n=12, Czech Republic n=93, Germany n=41 and Russia n=16) and cultivated on MacConkey agar with colistin (3.5 mgL?1). Enterobacteriaceae isolates with reduced susceptibility to colistin were tested for mcr-1 to mcr-8 genes by polymerase chain reaction and Sanger sequencing. Matrix-Assisted Laser Desorption/Ionization-Time of Flight mass spectrometry was used to identify selected isolates to species level. Susceptibility to a set of 18 antimicrobials was determined using disk diffusion method while broth dilution method was used to determine minimal inhibitory concentration (MIC) to colistin. Conjugation assay based on filter mating was carried to determine the conjugative property of mcr-1-carrying plasmid. Wholegenome sequencing (WGS) was performed using MiSeq (Illumina) platform to analyse the genetic content of selected isolates and investigate plasmids with mcr-1 gene. Overall, 13 (8%, n=168) Enterobacteriaceae isolates showed reduced susceptibility to colistin (Escherichia coli n=9, Klebsiella pneumoniae n=2 and Enterobacter cancerogenus n=2). The gene mcr-1 was positive in one E. coli isolate originating from a Black Kite sampled at Semenovod, Russia. Apart from resistance to colisitn (&gt; 4mg/L), the isolates showed susceptibility to all tested antibiotics. WGS analysis revealed mcr-1 positive isolate belonged to sequence type 2280. Sequencing and plasmid transfer experiments showed the location of mcr-1 on a conjugative IncI2 plasmid pDR164 (59891 bp, GenBank accession number MK542639). Plasmid pDR164 showed similarity to IncI2 plasmids found in E. coli isolates from swine, poultry, human and sewage in USA, Vietnam, Argentina and China, respectively. Comparison of pDR164 with other plasmids revealed high similarity in the backbone as well as the region carrying mcr. Unlike pDR164, plasmids from Vietnam, China and USA carried insertion sequence elements flanking the corresponding mcr gene. We report the first mcr-1 gene in E. coli from wildlife in Russia on a typical IncI2 plasmid. Considering the global spread of plasmid-encoded colistin resistance, migratory birds may represent a vehicle for dissemination of clinically-important antimicrobial resistance bacteria and genes.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

10606 - Microbiology

Projekt

<a href="/cs/project/NV18-09-00605" target="_blank" >NV18-09-00605: Enterobacteriaceae rezistentní ke karbapenemům a kolistinu: koncept jednoho zdraví v hodnocení humánních a jiných zdrojů rizikových mechanizmů antimikrobiální rezistence</a><br>

Návaznosti

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

Rok uplatnění

2019

Kód důvěrnosti údajů

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