Abstract
Pseudomonas aeruginosa is an opportunistic Gram-negative pathogen with an increase in the frequency of infections caused by multidrug resistant (MDR) and extensively drug resistant (XDR) strains, limiting the available therapeutic options. The most troublesome resistance is the acquisition and production of carbapenemases such as Verona integron-encoded metallo-β-lactamases (VIM), the most frequent and widespread, and the Klebsiella pneumoniae carbapenemases (KPC), which has continuously spread in the last decade. Its dissemination is linked to their location on mobile genetic elements (MGEs). In Colombia, VIM and KPC have been increasing in its frequency showing major successful dissemination. In this article, we molecularly characterized and analyzed the genetic context of blaVIM and blaKPC in carbapenem-resistant P. aeruginosa (CRPA) isolates from infected and colonized patients in two tertiary-care hospitals, one in Medellín and the other in a municipality close to Medellín, both areas with high carbapenemase endemicity in Colombia (2013–2015). Using whole-genome sequencing (WGS), we identified a remarkable variety of genetic backgrounds in these MDR P. aeruginosa isolates carrying blaKPC–2 and blaVIM–2. There were a diversity of class 1 integron and variations in the gene cassettes associated to blaVIM–2, as well as a possible event of spread of blaKPC–2 mediated by a plasmid that contained part of Tn4401b in one infection case. The dissemination of blaVIM–2 and blaKPC–2 in P. aeruginosa in this area in Colombia has been strongly influenced by successful international clones, carrying these genes and additional determinants of resistance on MGEs, accompanied by gene rearrangement under an antimicrobial selection pressure. These findings emphasize the need to implement control strategies based on rational antibiotic use.
Highlights
MATERIALS AND METHODSPseudomonas aeruginosa is an opportunistic Gram-negative pathogen especially in immunocompromised patients capable of causing a wide array of life-threatening infections
The 46 isolates of carbapenem-resistant P. aeruginosa (CRPA) from the two hospitals were recovered from 41 adult patients; 24 (58.5%) were infected and 17 (41.4%) were colonized, and 7 (17.1%) of them were localized in intensive care unit (ICU) at the time of sampling in the participating hospitals
This study provides new data supporting the genetic diversity and differences in the genetic context of blaVIM−2 and blaKPC−2 of multidrug resistant (MDR) P. aeruginosa isolates recovered from colonized and infected patients from two tertiary care hospitals, one in Medellín and the second located in a municipality close to Medellín, both areas with high carbapenemase endemicity in Colombia
Summary
Pseudomonas aeruginosa is an opportunistic Gram-negative pathogen especially in immunocompromised patients capable of causing a wide array of life-threatening infections. The study of genetic platforms of blaVIM and blaKPC in carbapenem-resistant P. aeruginosa (CRPA) that co-harbor genes conferring resistance to other antibiotics, as well as their wide diversity, is key to understand the role in the dissemination of such resistance determinants among clinical and environmental isolates (Cuzon et al, 2011; Naas et al, 2013; Correa et al, 2015; Abril et al, 2019). Easyfig was used to compare and visualize the backbone of different MGEs. The BLAST Ring Image Generator (BRIG) software (Alikhan et al, 2011) was applied to align the assembled reads of some sequenced clinical isolates to one reference plasmid carrying blaKPC. Comparison of clinical and epidemiological data was performed between colonized and infected patients, as well as between carbapenemase-producing P. aeruginosa (CPPA) and non-CPPA isolates. The sequence data for the isolates were submitted to the NCBI GenBank database under the BioProject number PRJNA391501
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