Characterization of the Complete Nucleotide Sequences of mcr-1-Encoding Plasmids From Enterobacterales Isolates in Retailed Raw Meat Products From the Czech Republic

Abstract

The aim of our study was to determine complete nucleotide sequence of mcr-1-carrying plasmids from Enterobacterales isolates recovered from domestic and imported raw retailed meat and compare them with plasmids available at the GenBank sequence database. A set of 16 plasmids originating from Escherichia coli (n = 13), Klebsiella pneumoniae (n = 2), and Citrobacter braakii (n = 1) were analyzed. In our previous study, data from whole genome sequencing showed that mcr-1 gene was located on plasmids of different incompatibility groups (IncHI2, IncI2, and IncX4). The IncI2 (n = 3) and IncX4 (n = 8) plasmids harbored mcr-1.1 gene only, whereas IncHI2 sequence type 4 plasmids (n = 5) carried large multidrug resistance (MDR) regions. MDR regions of IncHI2 plasmids included additional antimicrobial resistance genes conferring resistance to β-lactams (blaTEM−1), aminoglycosides [aadA1, aadA2, and aph(6)-Id], macrolides [mef (B)], tetracycline (tetA, tetR), and sulphonamides (sul1, sul2, and sul3). Likewise, IncHI2 plasmids carried several insertion sequences including IS1, IS3, IS26, IS1326, and ISApl1. In conclusion, our findings confirmed the involvement of IncX4, IncI2, and IncHI2 plasmids in the dissemination of mcr-1.1 gene in several environmental niches, as in samples of retail meat originating from different geographical regions. In contrast to IncX4 and IncI2, IncHI2 plasmids were more diverse and carried additional genes for resistance to heavy metals and multiple antimicrobials.

Characterisation of Early Positive mcr-1 Resistance Gene and Plasmidome in Escherichia coli Pathogenic Strains Associated with Variable Phylogroups under Colistin Selection

“…This stimulated the research and characterisation of plasmids carrying these virulence factors in relation to public health and genomic epidemiology. In fact, the mcr-1 element has been described on different types of plasmids, including IncI2, IncHI2, and IncX4 that are involved in the dissemination of the resistance gene in several environmental niches and they carry additional genes for resistance to heavy metals and multiple antimicrobials [ 14 , 15 ]. As part of a monitoring programme on antimicrobial susceptibility, 147 E. coli strains were isolated from faecal samples of cattle with diarrhoea and from milk-aliquots of cattle with mastitis in both France and Germany [ 16 ].…”

Detection of a New Resistance-Mediating Plasmid Chimera in a blaOXA-48-Positive Klebsiella pneumoniae Strain at a German University Hospital

“…IS66 is abundant in the worldwide-distributed K. pneumoniae sequence types ST258/ST512 [61]. IS3, using a two-step transposition mechanism to specifically insert into short palindromic repeated sequences, has been reported from K. pneumoniae in Europe and the USA [62][63][64]. IntIPac was described from a IncFIB plasmid found in a clinical Klebsiella variicola isolate in Hong Kong [65].…”

Potential sources and characteristic occurrence of mobile colistin resistance (mcr) gene-harbouring bacteria recovered from the poultry sector: a literature synthesis specific to high-income countries

“…Hence meat trade is a route for global dissemination of PMCR. The mcr-1 was on 34-45 kb IncX4, 55-60 kb IncI2, and 250-260 kb IncHI2 plasmids (with IncX4 predominating) in some of the isolates (Gelbíčová et al, 2019;Zelendova et al, 2021), indicating that IncI and IncH plasmids are the commonest drivers of PMCR resistance in the poultry environment in the countries from where the meats originated. There were nine other resistance genes (including ESBL and PMQR) in the strains (Gelbíčová et al, 2019;Zelendova et al, 2021) (Table 3), further supporting that poultry meat remains a potential source for spreading multi-to extensively drug-resistant organisms that would pose severe risk to public health.…”

“…The mcr-1 was on 34-45 kb IncX4, 55-60 kb IncI2, and 250-260 kb IncHI2 plasmids (with IncX4 predominating) in some of the isolates (Gelbíčová et al, 2019;Zelendova et al, 2021), indicating that IncI and IncH plasmids are the commonest drivers of PMCR resistance in the poultry environment in the countries from where the meats originated. There were nine other resistance genes (including ESBL and PMQR) in the strains (Gelbíčová et al, 2019;Zelendova et al, 2021) (Table 3), further supporting that poultry meat remains a potential source for spreading multi-to extensively drug-resistant organisms that would pose severe risk to public health. Conjugation was positive, and the microorganisms were extensively diverse, with the E. coli isolates belonging to 30 STs, including HiR pandemic ExPEC clones ST10, ST58, ST354, ST69, and 410 (Gelbíčová et al, 2019;Manges et al, 2019;Kubelova et al, 2021;Zelendova et al, 2021).…”

“…In the Czech Republic, 113 strains (110 E. coli and 3 K. pneumoniae) carrying mcr-1 were isolated from poultry meats sourced locally and imported into the country from Poland, Germany, and Brazil (Karpíšková et al, 2017;Gelbíčová et al, 2019;Gelbicova et al, 2020;Kubelova et al, 2021;Zelendova et al, 2021), suggesting that PMCR in Czechia poultry meat supply chain originate locally. Hence meat trade is a route for global dissemination of PMCR.…”