Conjugative IncHI2/HI2A plasmids harbouring mcr-9 in colistin-susceptible Escherichia coli isolated from diseased pigs in Japan

Colistin is a last resort antimicrobial used for the treatment of gram-negative bacterial infections. Plasmid-mediated colistin resistance (mcr) genes are a cause of global concern, and, thus far, mcr-1–10 have been identified. In a previous study, we screened mcr-1–5 in Escherichia coli derived from diseased pigs in Japan and reported a high prevalence of mcr-1, -3 and -5. However, the previous report on the prevalence of mcr genes was inaccurate. In the present study, we aimed to clarify the prevalence of all reported variants of mcr in E. coli derived from the diseased pigs, which were previously screened for mcr-1–5. Additionally, we also characterized the mcr-9-positive E. coli , which was detected in this study. We screened mcr in 120 E. coli strains from diseased pigs and mcr-positive E. coli and an mcr-carrying plasmid were also characterized. One mcr-9-positive colistin-susceptible E. coli strain was detected (0.8 %). Plasmid-mediated mcr-9 was transferred to E. coli ML4909 as the recipient strain, and it was located on IncHI2/HI2A plasmid p387_L with other antimicrobial resistance genes (ARGs). The region harbouring ARGs including mcr-9, was similar to that on the Klebsiella pneumoniae chromosome harbouring mcr-9 isolated in Japan. mcr-3, -5 and -9 were detected (4.2 %) in colistin-susceptible strains. mcr-9 was found to be disseminated via the plasmid IncHI2/HI2A p387_L and transferred and inserted into chromosomes via a transposon. Our results suggest that mcr genes should be monitored regularly, regardless of their susceptibility to colistin.

[1]  G. Valiakos,et al.  Colistin Resistant mcr Genes Prevalence in Livestock Animals (Swine, Bovine, Poultry) from a Multinational Perspective. A Systematic Review , 2021, Veterinary sciences.

[2]  Tatsuya Nakamura,et al.  Genomic Characterization of ESBL- and Carbapenemase-Positive Enterobacteriaceae Co-harboring mcr-9 in Japan , 2021, Frontiers in Microbiology.

[3]  C. Nakajima,et al.  Complete Genome Sequence of an mcr-10-Possessing Enterobacter roggenkampii Strain Isolated from a Dog in Japan , 2021, Microbiology resource announcements.

[4]  K. Holt,et al.  Silent spread of mobile colistin resistance gene mcr-9.1 on IncHI2 'superplasmids' in clinical carbapenem-resistant Enterobacterales. , 2021, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[5]  Tomomi Sato,et al.  Decreased colistin resistance and mcr-1 prevalence in pig-derived Escherichia coli in Japan after banning colistin as a feed additive. , 2021, Journal of global antimicrobial resistance.

[6]  D. Motooka,et al.  Genomic characterization of clinical Enterobacter roggenkampii co-harboring blaIMP-1- and blaGES-5-encoding IncP6 and mcr-9-encoding IncHI2 plasmids isolated in Japan. , 2020, Journal of global antimicrobial resistance.

[7]  Zhimin Guo,et al.  Molecular Detection of the mcr Genes by Multiplex PCR , 2020, Infection and drug resistance.

[8]  Z. Zong,et al.  Identification of novel mobile colistin resistance gene mcr-10 , 2020, Emerging microbes & infections.

[9]  A. Rajković,et al.  Global Burden of Colistin-Resistant Bacteria: Mobilized Colistin Resistance Genes Study (1980–2018) , 2019, Microorganisms.

[10]  K. Ganbarov,et al.  Molecular mechanisms related to colistin resistance in Enterobacteriaceae , 2019, Infection and drug resistance.

[11]  Yu Lin,et al.  Assembly of long, error-prone reads using repeat graphs , 2018, Nature Biotechnology.

[12]  J. Li,et al.  The rise and spread of mcr plasmid-mediated polymyxin resistance , 2019, Critical reviews in microbiology.

[13]  M. Wiedmann,et al.  Identification of Novel Mobilized Colistin Resistance Gene mcr-9 in a Multidrug-Resistant, Colistin-Susceptible Salmonella enterica Serotype Typhimurium Isolate , 2019, mBio.

[14]  Motomichi Takahashi,et al.  Inhibition effect of flavophospholipol on conjugative transfer of the extended-spectrum β-lactamase and vanA genes , 2018, The Journal of Antibiotics.

[15]  Toyotaka Sato,et al.  High prevalence of mcr-1, mcr-3 and mcr-5 in Escherichia coli derived from diseased pigs in Japan. , 2018, International journal of antimicrobial agents.

[16]  Qijing Zhang,et al.  Co-transfer of blaNDM-5 and mcr-1 by an IncX3–X4 hybrid plasmid in Escherichia coli , 2016, Nature Microbiology.

[17]  F. Beaudry,et al.  Resistance to colistin: what is the fate for this antibiotic in pig production? , 2016, International journal of antimicrobial agents.

[18]  Christina A. Cuomo,et al.  Pilon: An Integrated Tool for Comprehensive Microbial Variant Detection and Genome Assembly Improvement , 2014, PloS one.

[19]  Leena Salmela,et al.  LoRDEC: accurate and efficient long read error correction , 2014, Bioinform..

[20]  M. Ferraro Performance standards for antimicrobial susceptibility testing , 2001 .