Epidemic Territorial Spread of IncP-2-Type VIM-2 Carbapenemase-Encoding Megaplasmids in Nosocomial Pseudomonas aeruginosa Populations

Paweł Urbanowicz; Ibrahim Bitar; Radosław Izdebski; Anna Baraniak; Elżbieta Literacka; Jaroslav Hrabák; Marek Gniadkowski

doi:10.1128/AAC.02122-20

Epidemic Territorial Spread of IncP-2-Type VIM-2 Carbapenemase-Encoding Megaplasmids in Nosocomial Pseudomonas aeruginosa Populations

Antimicrob Agents Chemother. 2021 Mar 18;65(4):e02122-20. doi: 10.1128/AAC.02122-20. Print 2021 Mar 18.

Authors

Paweł Urbanowicz¹, Ibrahim Bitar², Radosław Izdebski¹, Anna Baraniak¹, Elżbieta Literacka³, Jaroslav Hrabák², Marek Gniadkowski⁴

Affiliations

¹ Department of Molecular Microbiology, National Medicines Institute, Warsaw, Poland.
² Biomedical Center, Faculty of Medicine in Plzen, Charles University, Plzen, Czech Republic.
³ Department of Epidemiology and Clinical Microbiology, The National Reference Centre for Susceptibility Testing, National Medicines Institute, Warsaw, Poland.
⁴ Department of Molecular Microbiology, National Medicines Institute, Warsaw, Poland m.gniadkowski@nil.gov.pl.

Abstract

In 2003 to 2004, the first five VIM-2 metallo-β-lactamase (MBL)-producing Pseudomonas aeruginosa (MPPA) isolates with an In4-like integron, In461 (aadB-bla_VIM-2-aadA6), on conjugative plasmids were identified in three hospitals in Poland. In 2005 to 2015, MPPA expanded much in the country, and as many as 80 isolates in a collection of 454 MPPA (∼18%) had In461, one of the two most common MBL-encoding integrons. The organisms occurred in 49 hospitals in 33 cities of 11/16 main administrative regions. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) classified them into 55 pulsotypes and 35 sequence types (STs), respectively, revealing their remarkable genetic diversity overall, with only a few small clonal clusters. S1 nuclease/hybridization assays and mating of 63 representative isolates showed that ∼85% of these had large In461-carrying plasmids, ∼350 to 550 kb, usually self-transmitting with high efficiency (∼10^-1 to 10^-2 per donor cell). The plasmids from 19 isolates were sequenced and subjected to structural and single-nucleotide-polymorphism (SNP)-based phylogenetic analysis. These formed a subgroup within a family of IncP-2-type megaplasmids, observed worldwide in pseudomonads from various environments and conferring resistance/tolerance to multiple stress factors, including antibiotics. Their microdiversity in Poland arose mainly from acquisition of different accessory fragments, as well as new resistance genes and multiplication of these. Short-read sequence and/or PCR mapping confirmed the In461-carrying plasmids in the remaining isolates to be the IncP-2 types. The study demonstrated a large-scale epidemic spread of multidrug resistance plasmids in P. aeruginosa populations, creating an epidemiological threat. It contributes to the knowledge on IncP-2 types, which are interesting research objects in resistance epidemiology, environmental microbiology, and biotechnology.

Keywords: IncP-2; MBL; Pseudomonas aeruginosa; VIM; carbapenemase; megaplasmid; metallo-β-lactamase; plasmid.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Anti-Bacterial Agents / pharmacology
Bacterial Proteins
Cross Infection* / epidemiology
Electrophoresis, Gel, Pulsed-Field
Epidemics*
Hospitals
Humans
Integrons / genetics
Multilocus Sequence Typing
Phylogeny
Poland
Pseudomonas Infections* / drug therapy
Pseudomonas Infections* / epidemiology
Pseudomonas aeruginosa / genetics
Pseudomonas aeruginosa / metabolism
beta-Lactamases / genetics
beta-Lactamases / metabolism

Substances

Anti-Bacterial Agents
Bacterial Proteins
beta-Lactamases
carbapenemase