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Background  The worldwide emergence and clonal spread of carbapenem-resistant  Acinetobacter baumannii  (CRAB) is of great concern. The aim of this nationwide study was to investigate the prevalence of CRAB isolates in Serbia and to characterize underlying resistance mechanisms and their genetic relatedness.    Methods  Non-redundant clinical samples obtained from hospitalized patients throughout Serbia were included in the prospective, observational, multicenter study conducted from January to June 2018. Samples were initially screened for the presence of  Acinetobacter baumannii-calcoaceticus  (Acb) complex using conventional bacteriological techniques. Acb complexes recovered from clinical samples obtained from inpatients with confirmed bacterial infections were further evaluated for the presence of  A. baumannii . Identification to the species level was done by the detection of the  bla  OXA-51  gene and  rpoB  gene sequence analysis. Susceptibility testing was done by disk diffusion and broth microdilution method. CRAB isolates were tested for the presence of acquired carbapenemases  (bla  OXA-24-like ,  bla  OXA-23-like,  bla  OXA-58-like ,  bla  OXA-143-like ,  bla  IMP ,  bla  VIM ,  bla  GIM ,  bla  SPM ,  bla  SIM ,  bla  NDM ) by PCR. Clonal relatedness was assessed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST).    Results  Acb complex was isolated in 280 out of 2401 clinical samples (11.6%). Overall,  A. baumannii  was identified in 237 out of 280 Acb complex (84.6%). CRAB prevalence was found to be 93.7% (237/222). The MIC 50 /MIC 90  for imipenem and meropenem were 8/> 32 μg/mL and 16/> 32 μg/mL, respectively. Although susceptibility was high for colistin (95.7%;  n  = 227) and tigecycline (75.1%;  n  = 178), ten isolates (4.3%) were classified as pandrug-resistant. The following carbapenemases-encoding genes were found: 98 (44.2%)  bla  OXA-24-like , 76 (34.5%)  bla  OXA-23-like , and 7 (3.2%)  bla  NDM-1 . PFGE analysis revealed six different clusters. MLST analysis identified three STs: ST2 ( n  = 13), ST492 ( n  = 14), and ST636 ( n  = 10). Obtained results evaluated that circulating CRAB clones in Serbia were as follows:  bla  OXA66 / bla  OXA23 /ST2 (32.4%),  bla  OXA66 / bla  OXA23 / bla  OXA72 /ST2 (2.7%),  bla  OXA66 / bla  OXA72 /ST492 (37.8%), and  bla  OXA66 / bla  OXA72 /ST636 (27.1%).    Conclusion  This study revealed extremely high proportions of carbapenem resistance among  A. baumannii  clinical isolates due to the emergence of  bla  OXA-72 ,  bla  OXA-23 , and  bla  NDM-1  genes among CRAB isolates in Serbia and their clonal propagation.

The first nationwide multicenter study of Acinetobacter baumannii recovered in Serbia: emergence of OXA-72, OXA-23 and NDM-1-producing isolates