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Colistin has become the last-line antimicrobial for the treatment of multidrug resistant (MDR)  Enterobacterales  in human medicine. To date, several colistin resistance genes have been described. Of them  mcr -1 is disseminated worldwide in  Escherichia coli  of human and animal origin. The aim of this study was to characterize  mcr -mediated resistance plasmids from  E. coli  of animal origin in Spain. From our strain collection, 70  E. coli  of pig origin collected between 2005 and 2014 (10 per year, except for years 2009–2010–2013) were randomly selected and screened for the presence of  mcr -genes. Additionally, 20  E. coli  isolated in 2011 from white storks ( Ciconia ciconia ) from the same urban household waste landfill associated colony were also included. Whole genome sequencing of  mcr -positive isolates was carried out on a MiSeq (Illumina). Hybrid whole genome sequencing strategy combining nanopore and Illumina technologies were performed in a selection of isolates to close the genomes and plasmids and identify the presence of antimicrobial resistance genes. Minimum inhibitory concentration (MIC) was used to assess the susceptibility to colistin. Mating experiments were carried out to evaluate transferability of the  mcr -genes. A total of 19  mcr -1 and one  mcr -4 positive isolates were detected, 15 from pigs distributed during the study period, and five from storks collected in 2011. No other  mcr -variants were found. The MICs for colistin ranged between 4 and >4 mg/L. High diversity of STs were detected among the  mcr-1  positive  E. coli  isolates, with only ST-10 shared between pigs and white storks. Except for one isolate, all were genotypic and phenotypically MDR, and five of them also harbored cephalosporin resistance genes ( bla  CTX–M–  14 ,  bla  SHV–  12 , and three  bla  CMY–  2 ).  mcr -1 genes were mobilizable by conjugation, associated with IncX4, IncHI2, and IncI2 plasmids. In our study,  mcr -1 genes have been circulating in pig farms since 2005 harbored by a variety of  E. coli  clones. Its persistence may be driven by co-selection since plasmids containing  mcr -1 also exhibit resistance to multiple drugs used in veterinary medicine. Furthermore, this is the first report of the presence of  mcr -1 gene in isolates from white storks in Spain. This finding highlights the potential importance of wildlife that forage at urban household waste landfills in the transmission and spread of colistin resistance genes.

mcr-Colistin Resistance Genes Mobilized by IncX4, IncHI2, and IncI2 Plasmids in Escherichia coli of Pigs and White Stork in Spain