Objectives: The aim of this study was to determine the prevalence of beta-lactamases in Acinetobacter spp. recovered from Lebanese patients over a 1-year period using phenotypic and molecular methods. Methods: A total of 100 non-duplicate consecutive Acinetobacter spp. isolates were collected from various clinical specimens. Antimicrobial susceptibility testing was performed by the disk diffusion method. Susceptibility to colistin, imipenem and meropenem was determined by broth microdilution. The beta-lactamase inhibitors phenylboronic acid, cloxacillin and ethylene diamine tetra-acetic acid (EDTA) were used for presumptive detection of KPC-type beta-lactamase, AmpC beta-lactamase and metallo-beta-lactamase (MBL), respectively. Simplex PCR was conducted for molecular detection of beta-lactamases. Trilocus PCR typing was performed to determine the clonality of the isolates. Results: Among the 100 Acinetobacter spp. isolates, 78% were resistant to imipenem and 84% to meropenem. Only one isolate was resistant to colistin by the microdilution method. Phenotypically, 23% of the isolates were presumptively diagnosed as producing extended-spectrum beta-lactamase (ESBL), 15% as producing KPC and 4% MBL, whilst 5% were diagnosed as overproducing AmpC beta-lactamase. The bla(OXA-51-like) gene was detected in 99% of isolates, bla(ADC) in 93%, bla(OXA-23-like) in 77% and bla(OXA-24/40-like) in 3%. Trilocus PCR identified 86% (82/95) of the Acinetobacter baumannii isolates as international clone II (IC II). Conclusions: A high rate of carbapenem resistance, with a predominance of OXA-23-like and IC II, was shown in this study. Moreover, the inhibitor-based method was shown not to be accurate for the prediction of carbapenemases in A. baumannii. (C) 2017 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.

INTRODUCTION:Increasing carbapenem resistance in Acinetobacter spp calls for the appraisal of alternative strategies in Acinetobacter spp infection therapy. This study aims at evaluating colistin-carbapenem combination against Acinetobacter spp using the checkerboard, Etest, and time-kill methods.METHODOLOGY:One hundred nonrepetitive Acinetobacter spp isolates were collected from patients admitted at the Saint-George-Hospital-University-Medical-Center over a one year period. The identification was performed using the API20NE and confirmed by the amplification of the blaOXA-51-like. Susceptibility to colistin, and carbapenems were determined using the Etest, microdilution methods and interpreted according to the CLSI, 2015. Detection of the carbapenemases was performed by PCR amplification method. Clonality was determined by the 3-Locus PCR-typing and ERIC-PCR methods. The synergistic potential of the combination was determined by calculating the Fractional-Inhibitory-Concentration-Index, which determines a synergistic, additive, indifferent or antagonistic effect.RESULTS:In our study (84%) of the isolates were carbapenem resistant. Only one strain showed resistance to colistin. (99%) and (77%) of the Acinetobacter spp isolates harbored blaOXA-51-like and blaOXA-23-like respectively. (86.2%) of the A.baumannii isolates pertained to the International Clone II. An additive effect of the colistin-carbapenem combination was determined using the 3 methods. A decrease of 2.6 and 2.8 folds in the MIC of colistin was showed in colistin-meropenem and colistin-imipenem, respectively (p < 0.001). The Colistin-meropenem showed better effects when compared to colistin-imipenem (p < 0.05). Only a few isolates showed a synergistic effect in the time-kill assay.CONCLUSION:Our study showed that the decrease in the MIC of colistin following colistin-carbapenem combination might be a promising antimicrobial approach for treating carbapenem-resistant Acinetobacter spp.

INTRODUCTION The worldwide emergence of antimicrobial resistance in Acinetobacter spp and their clonal dissemination call for the investigation into Acinetobacter spp epidemiology. METHODOLOGY 100 nonrepetitive Acinetobacter spp isolates were recovered from patients admitted at Saint- George-Hospital-University-Medical-Center-Beirut, in a one-year period. Identification of the isolates was determined by the API20NE and confirmed by PCR amplification of blaOXA-51-like. Susceptibility to carbapenems and colistin were determined by the microdilution method and interpreted according to the CLSI, 2015.The β lactamase inhibitors: PBA, EDTA, and Cloxacillin were used for the detection of KPC, MBL and AmpC, respectively. ESBL producers were detected whenever a keyhole effect was observed between 3rd generation cephalosporin and Augmentin®. Simplex PCR was conducted for the genotypic detection of β lactamases. ERIC and 3LST-PCR were performed to determine the clonality of the isolates. RESULTS Our findings showed that 84% were carbapenem resistant. Only one isolate was resistant to colistin. Phenotypically, 23 were ESBL, 15 KPC, 5 AmpC, and 4 MBL producers. PCR analysis showed that 99%, 93%, 77% and 3 % of the isolates harbored blaOXA-51-like, blaADC, blaOXA-23-like, and blaOXA-40-like, respectively. ERIC-PCR analysis showed that A.baumannii isolates were clustered in 19 possibly related and 30 closely related subtypes. The 3-LST-PCR showed that 86.2% of the A.baumannii isolates pertained to the ICII (international clone II). CONCLUSION Our study showed a predominance of OXA-23-like producers and dissemination of ICII. Inhibitor based method was shown not to be accurate for the prediction of carbapenemases in Acinetobacter spp. Infection control measures are needed for management of Acinetobacter spp infections.

The worldwide increase in the emergence of carbapenem resistant Acinetobacter baumannii (CRAB) calls for the investigation into alternative approaches for treatment. This study aims to evaluate colistin-carbapenem combinations against Acinetobacter spp., in order to potentially reduce the need for high concentrations of antibiotics in therapy. This study was conducted on 100 non-duplicate Acinetobacter isolates that were collected from different patients admitted at Saint George Hospital-University Medical Center in Beirut. The isolates were identified using API 20NE strips, which contain the necessary agents to cover a panel of biochemical tests, and confirmed by PCR amplification of blaOXA−51−like. Activities of colistin, meropenem and imipenem against Acinetobacter isolates were determined by ETEST and microdilution methods, and interpreted according to the guidelines of the Clinical and Laboratory Standards Institute. In addition, PCR amplifications of the most common beta lactamases contributing to carbapenem resistance were performed. Tri locus PCR–typing was also performed to determine the international clonality of the isolates. Checkerboard, ETEST and time kill curves were then performed to determine the effect of the colistin-carbapenem combinations. The synergistic potential of the combination was then determined by calculating the Fractional Inhibitory Concentration Index (FICI), which is an index that indicates additivity, synergism, or antagonism between the antimicrobial agents. In this study, 84% of the isolates were resistant to meropenem, 78% to imipenem, and only one strain was resistant to colistin. 79% of the isolates harbored blaOXA−23−like and pertained to the International Clone II. An additive effect for the colistin-carbapenem combination was observed using all three methods. The combination of colistin-meropenem showed better effects as compared to colistin-imipenem (p < 0.05). The colistin-meropenem and colistin-imipenem combinations also showed a decrease of 2.6 and 2.8-fold, respectively in the MIC of colistin (p < 0.001). Time kill assays additionally showed synergistic effects for a few isolates, and no bacterial re-growth was detected following a 24 h incubation. Our study showed that the combination of colistin with carbapenems could be a promising antimicrobial strategy in treating CRAB infections and potentially lowering colistin toxicity related to higher doses used in colistin monotherapy.