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Colistin is a last resort antibiotic used for the treatment of human infections associated with carbapenemase-producing Enterobacteriales. Here, we evaluated the occurrence of  mcr-1  and  -2  plasmid-mediated colistin resistance in colistin and/or carbapenem resistant human clinical Enterobacteriales and other gram-negative bacteria ( n  = 543) as well as third generation cephalosporin-resistant (3GCR)  Escherichia coli  isolates from poultry abattoir workers ( n  = 15) and poultry fecal samples ( n  = 92) collected from two geographically separate abattoirs in Romania. which revealed that  mcr-1  was present within four sequence types (STs): ST744 ( n  = 7), ST57 ( n  = 7), ST156 ( n  = 2), and ST10 ( n  = 1). Within STs, serotypes were conserved and, notably, all except one of the  mcr-1- positive isolates were found to exhibit fluoroquinolone-resistance (FQR) associated SNPs in both  gyrA  and  parC . While there were variations in genotypes, all isolates belonging to ST744, ST57, and ST156 were rich in resistance determinants, carrying aminoglycoside-modifying enzymes genes, sulfonamide resistance gene  bla  TEM–  1  as well as  bla  CMY–  2  AmpC β-lactamase resistance genes. They also exhibited high similarity in carriage of virulence genes; ST10, however, only carried the  mcr-1  gene. Whole genome sequencing (WGS) analysis also revealed that although the  mcr-1  gene was identified in a diverse population of  E. coli , two STs (ST57 and ST744) predominated and interestingly, were found in isolates across both abattoirs providing evidence for clonal transmission. Also, two main genomic contexts of  mcr-1  isolates were revealed with all ST57 isolates harboring the  mcr-1  gene between two copies of IS Apl1  (or the Tn 6330  transposon) whilst a common  mcr-1  containing scaffold, highly similar to IncX type  mcr-1 -bearing plasmids (pWI2-mcr, Accession number:  LT838201 ), was present among  mcr-1  isolates of varying phylogenetic backgrounds (ST10, ST744 and ST156). The high prevalence of the  mcr-1  gene in poultry  E. coli  isolates with co-resistance to cephalosporins and quinolones, in a country where antimicrobial use in food production species is poorly regulated, is concerning and the findings from this study should lead to better surveillance of antimicrobial resistance (AMR) in food-production animals in Romania.

Genetic Features of mcr-1 Mediated Colistin Resistance in CMY-2-Producing Escherichia coli From Romanian Poultry