ABSTRACT
BACKGROUND/AIMS: Colistin resistance is increasingly recognized among carbapenemase-producing Klebsiella pneumoniae isolates in several European regions. The current study documents the appearance of colistin resistance among KPC-2 and SHV-5-produning K. pneumoniae strains in Bulgaria. METHODS: Four colistin-resistant K. pneumoniae isolates were recovered from 2 patients hospitalized in the anesthesiology and resuscitation clinic of a tertiary care university hospital in Sofia, Bulgaria. Microbial identification and antimicrobial susceptibility testing was performed by Vitek 2 (Biomerieux, France). ß-Lactamase genes were amplified using a panel of primers for detection of all MBL-types, KPCs, plasmid-mediated AmpCs in single PCR reactions, OXA-type carbapenemases, extended-spectrum ß-lactamases (ESBLs) and TEM enzymes. The colistin-resistant mcr-1 gene was also investigated using previously described primers and conditions. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) were used to investigate clonality. RESULTS: The 4 K. pneumoniae isolates exhibited colistin MICs >16 mg/L and showed multidrug-resistant phenotypes, remaining intermediately susceptible only to gentamicin. They were clustered into a single PFGE clonal type and MLST assigned them to sequence type 258. All isolates possessed KPC-2 carbapenemase and SHV-5 ESBL. They were negative for the plasmid-mediated colistin-resistant mcr-1 gene, possibly implying an intrinsic mechanism of resistance. CONCLUSIONS: Although colistin use in Bulgaria only started moderately during 2014, the findings of the current study notify the appearance of colistin resistance among carbapenemase-producing Klebsiella species in another European region.
Subject(s)
Anti-Bacterial Agents/pharmacology , Colistin/pharmacology , Drug Resistance, Multiple, Bacterial/drug effects , Klebsiella Infections/diagnosis , Klebsiella pneumoniae/isolation & purification , Aged, 80 and over , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Bulgaria , DNA, Bacterial/isolation & purification , DNA, Bacterial/metabolism , Electrophoresis, Gel, Pulsed-Field , Female , Humans , Klebsiella Infections/microbiology , Klebsiella pneumoniae/enzymology , Klebsiella pneumoniae/genetics , Male , Middle Aged , Multilocus Sequence Typing , Tertiary Care Centers , beta-Lactamases/genetics , beta-Lactamases/metabolismABSTRACT
We report on the role of Acinetobacter radioresistens in a case of pneumonia in an elderly patient and describe the challenge of correct identification of this species. A tracheobronchial culture taken from a patient in a Bulgarian hospital yielded a pure culture of Gram-negative, lactose-non-fermenting bacilli on MacConkey agar. Genus and species identification was performed by biochemical tests and sequencing of the rpoB gene. Antimicrobial susceptibility testing and screening for blaOXA-like carbapenemase genes was done using microbroth dilution and PCR and sequencing, respectively. The bacillus growing on MacConkey agar was initially identified by biochemical tests as Acinetobacter baumannii complex. Sequencing of the rpoB gene finally identified A. radioresistens. The strain harboured the carbapenemase gene blaOXA-23 without insertion sequences upstream of this gene and was susceptible to imipenem and meropenem. In conclusion, detection of A. radioresistens remains a challenge for routine laboratory diagnostics without performance of molecular identification methods. Although A. radioresistens can be a causative agent of opportunistic infections, in the present case its involvement in the development of pneumonia is doubtful.
