ࡱ> @B?'`bjbjLULU ;(.?.?>zzzz 0.-------$.hN12-SSS--***SL-*S-**,, `bT6z(l,- .00.,2 *2,,2w-,* w-- * 0.SSSSdV$V4": Preliminary results of the multicentre study on carbapenem resistance of Acinetobacter baumannii in northern Croatia Abstract number: P704 Bedenic B., Vranic-Ladavac M., Budimir A., Rezo-Vranjes V., Sivec S., Sijak D., Vranes J., Beader N., Kalenic S., Bosnjak Z. Objectives: The aim of the study was to characterize the mechanisms of carbapenem resistance in A. baumannii isolates collected in the frames of a multicenter study and to study their molecular epidemiology. Material and Methods: In total 115 A. baumannii isolates were collected in 13 diagnostic laboratories located in Northern part of Croatia in 2009. Antibiotic susceptibilities were determined by broth microdilution method according to CLSI. E-test MBL strips were used for detection of metallo-b-lactamases (MBLs). PCR was used to detect the presence of oxacillinases (OXA-51, OXA-23, OXA-24 and OXA-58) and MBLs of VIM, IMP and SIM series. The genetic context of blaOXA-51 and blaOXA-58 genes was determined by PCR mapping with the primers for ISAbaI and IsabaIII combined with forward and reverse primers for blaOXA-51 and blaOXA-58. Genotyping of the isolates was performed by PFGE. Results: There were 97% of the strains resistant to piperacillin, 94% to ceftriaxone, cefotaxime and gentamicin, 91% to ciprofloxacin, 89% to ceftazidime, 68% to tobramycin, 60% to amikacin and 29% to ampicillin/sulbactam. No resistance to colistin was observed. Colistin was the most potent antibiotic with MIC90 of 2 mg/L. Twelve isolates were found to be resistant to both carbapenems. Nine of them were shown to possess blaOXA58-like gene and three blaOXA-40 like gene. The strains susceptible to carbapenems were positive only for naturally ocurring OXA-51 b-lactamase whereas eight isolates with intermediate susceptibility to one or both carbapenems were shown to possess ISAbaI insertion sequence upstream of blaOXA-51 gene. No MBLs were found. Only OXA-58 b-lactamase was inhibited by sodium chloride. Chromosomal AmpC b-lactamases did not affect the susceptibility to carbapenems. None of the isolates exhibited significant synergy between meropenem and CCCP, indicating that overexpression of proton-gradient depended efflux pumps did not contribute to the meropenem resistance. The strains producing only OXA-51 b-lactamase were found to belong to EU clone I whereas OXA-40 and OXA-58 producers were asigned to EU clone II. Three strains positive for blaOXA-40 like genes were clonally related. Conclusions: The emergence of carbapenem resistance in A. baumannii in Northern Croatia is associated with the production of acquired oxacillinases belonging to OXA-58 and OXA-40 like groups. Carbapenem resistant isolates producing acquired oxacillinases belonged to EU clone II. Session Details Date: 07/05/2011 Time: 00:00-00:00 Session name: Abstracts of 21st ECCMID / 27th ICC Subject: Location: Milan, Italy, 7 - 10 May 2011 Presentation type: Klebsiella pneumoniae harbouring NDM1 metallo-lactamase isolated in Croatia Abstract number: P648 Mazzariol A., Bosnjak Z., Ballarini P., Budimir A., Bedenic B., Kalenic S., Cornaglia G. Objectives: A K. pneumoniae strain, resistant to carbapenems, was isolated in May 2009, from a blood culture of a 40-years-old man, recovered in a surgical intensive care unit of the Clinical Hospital Center in Zagreb, Croatia. The patient had been transferred from another hospital of a neighbouring country after 5 days of hospitalization due to a car accident. The clinical history mentioned antibiotic treatment not including carbapenems (gentamicine, metronidazole and ceftriaxone), and no link to India or Pakistan. We investigated the carbapenem resistance mechanism. Materials and Methods: Antimicrobial susceptibility testing was performed by both Vitek2 and microdiluition, and interpreted according to the latest EUCAST documents. The presence of a carbapenemase was investigated by means of an MBL-etest, hydrolysis of carbapenems and EDTA inhibition of hydrolysis. PCR and sequencing were carried out by standard procedures. Resistance was transferred by conjugation to E. coli J53. Results: The strain was resistant to imipenem and meropenem (MICs 16 mg/L and 64 mg/L, respectively), and to all 3G-cephalosporins (MICs >128 mg/L), and susceptible to ciprofloxacin (MIC 0.25 mg/L) It also harboured a qnrA and a ctx-m-15 genes. The presence of an MBL was suggested by the MBL Etest and confirmed spectrophotometric by EDTA-inhibited imipenem hydrolysis. PCR performed by using primers specific for blaIMP, blaVIM, blaSPM, blaGIM, and blaSIM, did not yield any product. A product obtained with NDM primers was confirmed, after sequencing, to code for an NDM-1 enzyme. The resistance to carbapenems and 3G-cephalosporins was transferred to E. coli J53 by conjugation. Conclusions: To our knowledge this is the first report of an NDM-1-producing K. pneumoniae in Croatia and in the Balkans. The patient had no apparent link to the Indian subcontinent and the strain was isolated before the recent outbreak of NDM-1 producing Enterobacteriaceae. Although several cases reported by a recent pan-European survey showed a clear link to the Balkan region, this is the first documented report of NDM-1 from that area, thus confirming a possible multifocal spread of this enzyme and prompting for a widespread epidemiological surveillance. Session Details Date: 07/05/2011 Time: 00:00-00:00 Session name: Abstracts of 21st ECCMID / 27th ICC Subject: Location: Milan, Italy, 7 - 10 May 2011 Presentation type: wy hMho hMhMhMuv 0;dwxgdMxyz789:yz )=p{gdMgdM90&P 1h:p*. 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