title: VIM-2 beta-lactamase in Pseudomonas aeruginosa isolates from Zagreb, Croatia.
abstract: The aim of this investigation was to characterize metallo-beta-lactamases (MBLs) in Pseudomonas aeruginosa isolates from Zagreb, Croatia. One hundred P. aeruginosa isolates with reduced susceptibility to either imipenem or meropenem were tested for the production of MBLs by MBL-Etest. The susceptibility to a wide range of antibiotics was determined by broth microdilution method. The presence of bla(MBL) genes was detected by polymerase chain reaction (PCR). Hydrolysis of 0.1 mM imipenem by crude enzyme preparations of beta-lactamases was monitored by UV spectrophotometer. Outer membrane proteins were prepared and analysed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Six out of 100 isolates were positive for MBLs by Etest. All strains were resistant to gentamicin, ceftazidime and cefotaxime, and all except 1 were resistant to imipenem. Six strains positive for MBLs by Etest were identified as VIM MBL-producers by PCR. Sequencing of bla(VIM) genes revealed the production of VIM-2 beta-lactamase in all 6 strains. This investigation proved the occurrence of VIM-2 beta-lactamase among P. aeruginosa strains from Zagreb, Croatia. VIM-2 beta-lactamase with similar properties has previously been described in another region of Croatia and in Italy, France, Spain, Greece, Taiwan and South Korea, suggesting that this type of enzyme is widespread in the Mediterranean region of Europe and in the Far East.
authors.affiliation: Department of Clinical and Molecular Microbiology, Clinical Hospital Centre Zagreb, Zagreb, Croatia.
authors.forename: Zrinka
authors.initials: Z
authors.lastname: Bosnjak
authors.forename: Branka
authors.initials: B
authors.lastname: Bedenić
authors.forename: Annarita
authors.initials: A
authors.lastname: Mazzariol
authors.forename: Neda
authors.initials: N
authors.lastname: Jarza-Davila
authors.forename: Sandra
authors.initials: S
authors.lastname: Suto
authors.forename: Smilja
authors.initials: S
authors.lastname: Kalenić
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D017523:Croatia; D004269:DNA, Bacterial / Q000235:genetics; D006801:Humans; D006868:Hydrolysis; D015378:Imipenem / Q000378:metabolism / Q000494:pharmacology; D000077731:Meropenem; D008826:Microbial Sensitivity Tests; D016133:Polymerase Chain Reaction; D011552:Pseudomonas Infections / Q000382:microbiology*; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D013053:Spectrophotometry; D013845:Thienamycins / Q000494:pharmacology; D018440:beta-Lactam Resistance; D001618:beta-Lactamases / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D013845:Thienamycins; D015378:Imipenem; C425108:beta-lactamase bla(vim-2); D001618:beta-Lactamases; D000077731:Meropenem