Characterization of Isolates Associated with Emergence of OXA-48-ProducingKlebsiella pneumoniaein Croatia

Abstract

Here, we report a retrospective study conducted to elucidate emergence, epidemiology, and molecular mechanisms of resistance underlying the early spread of OXA-48 carbapenemase-producing Enterobacteriaceae in Croatia. Retrospective screening for OXA-48 producers was performed on a collection of 296 nonrepetitive, carbapenem-nonsusceptible enterobacterial isolates collected from January 2011 to December 2012 from 40 participating centers in Croatia. Antimicrobial susceptibility profiles and production of carbapenemases were assessed phenotypically. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing were used for epidemiological analysis. Resistance genes were characterized by polymerase chain reaction (PCR) and sequencing. Plasmid localization of bla in isolates and transconjugants was investigated by S1-PFGE and Southern hybridization. PCR mapping was used for identification of genetic platform surrounding bla. Out of 296 carbapenem-nonsusceptible isolates, bla gene was detected in 12 Klebsiella pneumoniae isolates. All OXA-48-producing isolates showed varying resistance to carbapenems and 11 were multidrug resistant. All coproduced additional beta-lactamases, including CTX-M-15, which was detected in eight isolates. Isolates were delineated in five clonal types by PFGE corresponding to five sequence types (STs) assigned ST15, ST16, ST37, ST528, and ST1418. All OXA-48 isolates conjugated successfully and other resistance determinants were not cotransferred. bla was carried on a ∼60 kb IncL/M plasmid and was detected within Tn1999.2 composite transposon. OXA-48, a class D carbapenemase, is emerging as a potentially significant contributor among carbapenem-resistant Enterobacteriaceae in Croatia, alongside class A and B carbapenemases. Polyclonal genetic background of K. pneumoniae isolates carrying ∼60 kb incL/M plasmid indicates that dissemination of the bla gene is not driven exclusively by the spread of a single clone.

OXA-48 Carbapenemase in Klebsiella pneumoniae Sequence Type 307 in Ecuador

“…We recently reported a strain of Raoutella ornithinolytica that harbored the blaOXA-48-like gene with no plasmid characterization that was recovered after this strain [59], and to the best of our knowledge, this is the first description of carrying OXA-48-like carbapenemase in an IncL plasmid isolated in Ecuador. Interestingly, our patient came from Croatia were this mechanism of resistance was reported previously [60][61][62], indicating that contagion with this strain most likely happened in Europe. The implications of globalization and free travel are now being revealed with the fast spread of the SARS-CoV-19, but in reality, this is nothing new and it has long been the case with mechanisms of resistance.…”

Whole genome sequencing characterization of Slovenian carbapenem-resistant Klebsiella pneumoniae, including OXA-48 and NDM-1 producing outbreak isolates

“…Moreover, four isolates of ST15, a pandemic clone [36], also detected in our neighbouring countries Croatia, Austria, and Hungary and associated with OXA-48, NDM-1 and CTX-M-15 producers [37][38][39], were detected in our selection, forming an unexpected outbreak. All four isolates shared the same beta-lactamase resistance pattern; however, additional epidemiological investigations have failed to reveal any clear connection.…”

A Novel Alkaliphilic Streptomyces Inhibits ESKAPE Pathogens

“…ESKAPE pathogens listed below and other ATCC strains listed were provided by Dr. M. Jelić ( Jelic et al, 2016 , 2017 ).…”

“…The ESKAPE pathogens were clinical isolates collected through regular hospital activities. Species identification was performed using standard biochemical methods (tests) and the VITEK 2 system (bioMérieux, France) ( Jelic et al, 2016 , 2017 ). Isolates were assigned unique isolate IDs and subsequently anonymized with designations a, b, c, etc.…”

“…If all solutions fail, infections by ESKAPE pathogens can lead to death of the patient and a spread of multi-resistant strains. Antibacterial resistance has now been detected for nearly all new antibiotics, even those of last resort ( Jelic et al, 2017 ; Li and Webster, 2018 ). As a consequence, the WHO have created an urgent priority list for discovery of new antibiotics ( Tacconelli et al, 2018 ).…”