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• W4366779230 abstract "We report the isolation of a multidrug-resistant Acinetobacter bereziniae isolate harbouring a plasmid blaNDM-1 in central Sydney, Australia from a patient who had travelled to the Philippines. While carbapenem-resistant Acinetobacter baumannii complex (e.g., A. baumannii, A. calcoaceticus, A. baumannii, A. dijkshoorniae, A. nosocomialis, A. pittii and A. seifertii) (CRAB) group have been previously described, carrying the blaNDM-1,1Music M.S. Hrenovic J. Goic-Barisic I. et al.Emission of extensively-drug-resistant Acinetobacter baumannii from hospital settings to the natural environment.J Hosp Infect. 2017; 96: 323-327Abstract Full Text Full Text PDF PubMed Scopus (53) Google Scholar, 2Australian Commission on Safety and Quality in Health CareNational Alert System for Critical Antimicrobial Resistances (CARAlert): Laboratory Handbook. ACSQHC, Sydney2022https://www.safetyandquality.gov.au/publications-and-resources/resource-library/caralert-laboratory-handbook-0Google Scholar, 3Dortet L. Poirel L. Nordmann P. Worldwide dissemination of the NDM-type carbapenemases in Gram-negative bacteria.Biomed Res Int. 2014; 2014249856Crossref PubMed Scopus (357) Google Scholar only a few cases of A. bereziniae clinical isolation have been described worldwide carrying such resistance.4Pavoni T. Chaga G. Detection of an NDM-1-producing Acinetobacter bereziniae strain in Brazil.J Glob Antimicrob Resist. 2015; 3: 147-148Crossref PubMed Scopus (13) Google Scholar,5Brovedan M. Marchiaro P.M. Moran-Barrio J. et al.Complete sequence of a blaNDM-1-harboring plasmid in an Acinetobacter bereziniae clinical strain isolated in Argentina.Antimicrob Agents Chemother. 2015; 59: 6667-6669Crossref PubMed Scopus (14) Google Scholar Acinetobacter berezinae is an environmental Acinetobacter species of the non-baumanii complex, with some routine medical biochemical databases such as the Vitek 2 XL unable to identify it correctly. Conversely, matrix-assisted laser desorption ionisation time of flight mass spectrometry (MALDI-TOF MS) offers a rapid and convenient identification method for routine laboratories,6Lee S.Y. Shin J.H. Kim S.H. et al.Evaluation of matrix-assisted laser desorption ionization-time of flight mass spectrometry-based VITEK MS system for the identification of Acinetobacter species from blood cultures: comparison with VITEK 2 and MicroScan systems.Ann Lab Med. 2015; 35: 62-68Crossref PubMed Scopus (15) Google Scholar and has been shown here to accurately identify an A. bereziniae isolate as confirmed by molecular methods. Furthermore, A. bereziniae harbouring a plasmid blaNDM-1 described here highlights the potential role of non-baumannii Acinetobacter disseminating antibiotic resistance genes through hospital environments.7Atrouni A.A. Joly-Guillou M.-L. Hamze M. Kempf M. Reservoirs of non-baumannii Acinetobacter species.Front Microbiol. 2016; 7: 1-7Crossref PubMed Scopus (132) Google Scholar The Acinetobacter bereziniae isolate carrying the metallo-β-lactamase resistance gene was isolated from the rectal swabs collected during infection control screening after the patient's recent hospitalisation in the Philippines. The organism did not grow on ESBL Brilliance agar (ThermoFisher, Australia) but grew on CRE Brilliance agar (ThermoFisher) as clear, slightly pigmented yellow colonies incubated 24–48 h at room temperature as instructed by the manufacturer (Fig. 1). The organism growing on the selective CRE Brilliance agar media was a Gram-negative coccobacillus by Gram stain and sub-cultured on horse blood agar (ThermoFisher) for further identification and susceptibility testing. The organism was first identified as Acinetobacter bereziniae by MALDI-TOF MS (Bruker, Australia) with a score of 2.53. This was a good reliable result using this identification system since a score value ≥2 indicated species identification; a score value between 1.7 and 1.9 indicated genus identification, and a score value <1.7 indicated no identification. Identification using the Vitek 2 XL (bioMerieux, Australia) system was also performed by using the GNI ID card with reference code 21341 (bioMerieux). After 9.92 hours, the automated identification system gave an excellent identification as Acinetobacter lwoffii with a probability of 99% with a Bionumber of 0001011101500102. The identification as Acinetobacter bereziniae was later confirmed using 16S rRNA sequencing. Results showed a 100% match using sequence data assembled and compared with previously reported sequence by using the basic local alignment search tool (BLAST) of the National Centre for Biotechnology Information (NCBI) database. On further testing the RAPIDEC CARBA NP (bioMerieux) was positive for the isolate which was indicative of the presence of a carbapenemase gene. Molecular testing using the gene Xpert Carba-R (Cepheid, Australia), a real-time polymerase chain reaction assay for rapid detection and differentiation of five genes (blaKPC, blaVIM, blaOXA-48, blaIMP-1, blaNDM) revealed the presence of a blaNDM carbapenemase gene. The isolate was later referred to a reference laboratory, the Institute of Clinical Pathology and Medical Research (ICPMR) in Westmead Hospital, Sydney, for whole genome sequencing which confirmed the presence of blaNDM-1 and further classified it as ST1318. Carbapenem resistance and further susceptibility of the organism to other antibiotics were confirmed using Vitek 2 XL, Etest strips (bioMerieux) and disc-diffusion (Thermofisher) susceptibility testing using both European Committee on Antimicrobial Susceptibility Testing (EUCAST) and Clinical and Laboratory Standards Institute (CLSI) guidelines.8Clinical and Laboratory Standards InstitutePerformance Standards for Antimicrobial Susceptibility Testing.32nd ed. CLSI Document M100. CLSI, Wayne, PA2022Google Scholar,9European Committee on Antimicrobial Susceptibility TestingBreakpoint tables for interpretation of MICs and zone diameters.2022https://www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/Breakpoint_tables/v_12.0_Breakpoint_Tables.pdfGoogle Scholar Colistin susceptibility was confirmed by broth microdilution (BMD) using the MICRONAUT MIC-Strip (MERLIN Diagnostika, Dutec, Australia). BMD quality control was performed with both E. coli ATCC 25922 and E. coli NCTC 13846 (mcr-1 positive). Acinetobacter bereziniae antibiotic susceptibility results are shown in Table 1.Table 1Antibiotic resistant profile of the Acinetobacter bereziniae carrying the blaNDM-1 geneAntibioticMICEUCASTCLSIMeropenemaMIC determined by Etest strips./bMIC determined by Vitek 2XL ASTN246 card.8IRImipenemaMIC determined by Etest strips.6RRCiprofloxacinaMIC determined by Etest strips.>34RRPiperacillin/tazobactamaMIC determined by Etest strips.12–ICefepimebMIC determined by Vitek 2XL ASTN246 card.>64–RGentamicinbMIC determined by Vitek 2XL ASTN246 card.4SSAmikacinaMIC determined by Etest strips.32RRTetracylineaMIC determined by Etest strips.24–RTigecyclineaMIC determined by Etest strips.1––AztreonamaMIC determined by Etest strips.>64––ColistincMIC confirmed by BMD quality control performed with both E. coli ATCC 25922 and E. coli NCTC 13846 (mcr-1 positive).2SSCLSI, Clinical and Laboratory Standards Institute; EUCAST, European Committee on Antimicrobial Susceptibility Testing; MIC, minimum inhibitory concentration.Interpretations: I, intermediate; R, resistant; S, susceptible; –, no breakpoints.a MIC determined by Etest strips.b MIC determined by Vitek 2XL ASTN246 card.c MIC confirmed by BMD quality control performed with both E. coli ATCC 25922 and E. coli NCTC 13846 (mcr-1 positive). Open table in a new tab CLSI, Clinical and Laboratory Standards Institute; EUCAST, European Committee on Antimicrobial Susceptibility Testing; MIC, minimum inhibitory concentration. Interpretations: I, intermediate; R, resistant; S, susceptible; –, no breakpoints. Acinetobacter species are ubiquitous in nature, and their ability to survive in many ecological niches is worrisome, especially when carrying multidrug-resistant genes in a hospital environment. Acinetobacter bereziniae, also referred to as Acinetobacter genospecies 10, has been isolated in sewage in Denmark, environmental surfaces in Korea, vegetables in Hong Kong and UK, meat and animals in Lebanon, as well as human skin in Germany and Hong Kong.7Atrouni A.A. Joly-Guillou M.-L. Hamze M. Kempf M. Reservoirs of non-baumannii Acinetobacter species.Front Microbiol. 2016; 7: 1-7Crossref PubMed Scopus (132) Google Scholar The first A. bereziniae carrying the blaNDM-1 gene was reported in Brazil in 2014 from a 69-year-old HIV-positive man admitted to ICU due to respiratory illness.4Pavoni T. Chaga G. Detection of an NDM-1-producing Acinetobacter bereziniae strain in Brazil.J Glob Antimicrob Resist. 2015; 3: 147-148Crossref PubMed Scopus (13) Google Scholar Other cases include a bacteraemia in Argentina from a 53-year-old patient who had undergone chemotherapy due to leukaemia,5Brovedan M. Marchiaro P.M. Moran-Barrio J. et al.Complete sequence of a blaNDM-1-harboring plasmid in an Acinetobacter bereziniae clinical strain isolated in Argentina.Antimicrob Agents Chemother. 2015; 59: 6667-6669Crossref PubMed Scopus (14) Google Scholar and cases of this organism carrying other metallo-β-lactamase resistances such as blaNDM, blaIMP and blaVIM genes have also been reported in Japan.10Yamamoto M. Nagao M. Matsumura Y. et al.Regional dissemination of Acinetobacter species harbouring metallo-β-lactamase genes in Japan.Clin Microbiol Infect. 2013; 19: 729-736Abstract Full Text Full Text PDF PubMed Scopus (25) Google Scholar,11Grosso F. Silva L. Sousa C. et al.Extending the reservoir of blaIMP5 the emerging pathogen Acinetobacter bereziniae.Future Microbiol. 2015; 10: 1609-1613Crossref PubMed Scopus (10) Google Scholar To our knowledge no one has previously reported the isolation of A. bereziniae carrying metallo-β-lactamase-resistant blaNDM-1 within the central Sydney region. Phenotypic identification of non-Acinetobacter species is problematic and challenging, as shown here with A. bereziniae misidentified as A. lwoffi from our Vitek 2 identification results, and also described by other studies with the Vitek MS and MicroScan.6Lee S.Y. Shin J.H. Kim S.H. et al.Evaluation of matrix-assisted laser desorption ionization-time of flight mass spectrometry-based VITEK MS system for the identification of Acinetobacter species from blood cultures: comparison with VITEK 2 and MicroScan systems.Ann Lab Med. 2015; 35: 62-68Crossref PubMed Scopus (15) Google Scholar MALDI-TOF MS offers a rapid, convenient identification method for diagnostic laboratories, and can be coupled with other molecular techniques such as 16S rRNA sequencing, rpoB, ARDRA, SDS-PAGE, ribotyping, DNA-DNA hybridisation, RAPD or whole genome sequencing6Lee S.Y. Shin J.H. Kim S.H. et al.Evaluation of matrix-assisted laser desorption ionization-time of flight mass spectrometry-based VITEK MS system for the identification of Acinetobacter species from blood cultures: comparison with VITEK 2 and MicroScan systems.Ann Lab Med. 2015; 35: 62-68Crossref PubMed Scopus (15) Google Scholar,7Atrouni A.A. Joly-Guillou M.-L. Hamze M. Kempf M. Reservoirs of non-baumannii Acinetobacter species.Front Microbiol. 2016; 7: 1-7Crossref PubMed Scopus (132) Google Scholar provided by reference laboratories. It should be noted that the current National Alert System for Critical Antimicrobial Resistances (CARAlert) handbook2Australian Commission on Safety and Quality in Health CareNational Alert System for Critical Antimicrobial Resistances (CARAlert): Laboratory Handbook. ACSQHC, Sydney2022https://www.safetyandquality.gov.au/publications-and-resources/resource-library/caralert-laboratory-handbook-0Google Scholar in Australia lists Acinetobacter baumannii complex species such as A. calcoaceticus, A. baumannii, A. dijkshoorniae, A. nosocomialis, A. pittii and A. seifertii, as potential organisms carrying carbapenem resistance; however, there is also the worrisome potential role of other non-baumannii Acinetobacter species such as A. bereziniae to carry carbapenem-resistant genes which have implications in infection control in hospital environments. Acinetobacter bereziniae carrying the blaNDM-1 have been shown to carry ISAba elements very similar to multidrug-resistant A. baumannii isolates, with the potential ability to mobilise as a whole and act as reservoirs of blaNDM genes which may contribute to their spreading among clinically relevant Enterobacterales.12Walsh T.R. Weeks J. Livermore D.M. Toleman M.A. Dissemination of NDM-1 positive bacteria in the New Delhi environment and its implications for human health: an environmental point prevalence study.Lancet Infect Dis. 2015; 11: 355-362Abstract Full Text Full Text PDF Scopus (975) Google Scholar We thank Dr Andrew Ginn, the Institute of Clinical Pathology and Medical Research (ICPMR), Westmead Hospital, Sydney for whole genome sequencing of the isolate and confirming the presence of blaNDM-1. The authors state there are no conflicts of interest to disclose." @default.
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• W4366779230 title "Isolation of Acinetobacter bereziniae harbouring plasmid blaNDM-1 in central Sydney, Australia" @default.
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