Ann Lab Med.  2018 Nov;38(6):555-562. 10.3343/alm.2018.38.6.555.

Detection of mcr-1 Plasmids in Enterobacteriaceae Isolates From Human Specimens: Comparison With Those in Escherichia coli Isolates From Livestock in Korea

• Yoon EJ
• Hong JS
• Yang JW
• Lee KJ
• Lee H
• Jeong SH

• KMID: 2429121
• DOI: http://doi.org/10.3343/alm.2018.38.6.555

Abstract

BACKGROUND
The emerging mobile colistin resistance gene, mcr-1, is an ongoing worldwide concern and an evaluation of clinical isolates harboring this gene is required in Korea. We investigated mcr-1-possessing Enterobacteriaceae among Enterobacteriaceae strains isolated in Korea, and compared the genetic details of the plasmids with those in Escherichia coli isolates from livestock.
METHODS
Among 9,396 Enterobacteriaceae clinical isolates collected between 2010 and 2015, 1,347 (14.3%) strains were resistant to colistin and those were screened for mcr-1 by PCR. Colistin minimum inhibitory concentrations (MICs) were determined by microdilution, and conjugal transfer of the mcr-1-harboring plasmids was assessed by direct mating. Whole genomes of three mcr-1-positive Enterobacteriaceae clinical isolates and 11 livestock-origin mcr-1-positive E. coli isolates were sequenced.
RESULTS
Two E. coli and one Enterobacter aerogenes clinical isolates carried carried IncI2 plasmids harboring mcr-1, which conferred colistin resistance (E. coli MIC, 4 mg/L; E. aerogenes MIC, 32 mg/L). The strains possessed the complete conjugal machinery except for E. aerogenes harboring a truncated prepilin peptidase. The E. coli plasmid transferred more efficiently to E. coli than to Klebsiella pneumoniae or Enterobacter cloacae recipients. Among the three bacterial hosts, the colistin MIC was the highest for E. coli owing to the higher mcr-1-plasmid copy number and mcr-1 expression levels. Ten mcr-1-positive chicken-origin E. coli strains also possessed mcr-1-harboring IncI2 plasmids closely related to that in the clinical E. aerogenes isolate, and the remaining one porcine-origin E. coli possessed an mcr-1-harboring IncX4 plasmid.
CONCLUSIONS
mcr-1-harboring IncI2 plasmids were identified in clinical Enterobacteriaceae isolates. These plasmids were closely associated with those in chicken-origin E. coli strains in Korea, supporting the concept of mcr-1 dissemination between humans and livestock.

Keyword

mcr-1; Colistin resistance; Enterobacteriaceae; IncI2 plasmid

MeSH Terms

Colistin
Enterobacter aerogenes
Enterobacter cloacae
Enterobacteriaceae*
Escherichia coli*
Escherichia*
Genome
Humans*
Klebsiella pneumoniae
Korea*
Livestock*
Microbial Sensitivity Tests
Plasmids*
Polymerase Chain Reaction
Colistin

Figure

• 

  Fig. 1 Comparative analyses of the mcr-1-plasmids from clinical Enterobacteriaceae strains and livestock-origin Escherichia coli strains. (A) Left, molecular phylogeny was conducted by neighbor joining analysis of nucleotide sequences (324 bp) of the replication origin of the mcr-1-plasmids. Multiple sequence alignments were performed with MUSCLE v3.8, and the phylogenetic tree was reconstructed using the distance method implemented in the BioNJ program [34]. Plasmid names are indicated in each taxon along with the size in brackets. Strain name and GenBank accession number are indicated below the plasmid name, if available. Color codes: Black, clinical Enterobacteriaceae strains; blue, E. coli from healthy chickens; red, E. coli from chicken carcasses; and green, E. coli from diseased pig. Right, schematic representation of plasmid structures. The sequence of each plasmid was aligned using BlastN and compared using the Artemis Comparison Tool. Highly-conserved regions (>96% nucleic acid identity) are indicated in red, and moderately conserved regions (>92% nucleic acid identity) are indicated in blue. Open arrows, open reading frames; blue, the replication origin; red, antimicrobial resistance; yellow, plasmid backbone; orange, transposases; and green, plasmid transfer. (B) The yellow arrow indicates the site-specific recombinase rci gene, and the green arrow indicates the pilUs assembly pilV gene. Black arrowheads represent the six 19-bp repeats. Open reading frames in the direction of translation are indicated by arrows.

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