Sir, Distinct clonal lineages of Acinetobacter baumannii that produce OXA carbapenemases have successfully disseminated in hospitals worldwide.1 In contrast, there are few reports about acquired OXA carbapenemases (e.g. OXA-23, OXA-58 and OXA-24/40) in A. baumannii from animal sources. Such findings in pets are so far limited to A. baumannii with OXA-23 carbapenemase in cats and dogs in Portugal, Germany and France.2–4 This is, to our knowledge, the first report about a carbapenem-resistant A. baumannii isolate with acquired carbapenemase OXA-72 recovered from a domestic grey parrot. The isolate, identified by using MALDI-TOF MS (Bruker Daltoniks, Bremen, Germany), was obtained from a choanal swab of a domestic grey parrot in mixed bacterial culture. The bird was presented to a veterinary clinic in Luxembourg in 2016 due to progressive dyspnoea and nasal discharge without prior antibiotic treatment. Antimicrobial susceptibilities of the A. baumannii isolate were determined by the VITEK 2 system (cards AST-GN38 and AST-N248) for ampicillin, amoxicillin/clavulanate, piperacillin, cefalexin, cefotaxime, ceftazidime, cefepime, ceftiofur, cefpirome, aztreonam, imipenem, meropenem, amikacin, gentamicin, tobramycin, ciprofloxacin, moxifloxacin, enrofloxacin, marbofloxacin, tetracycline, tigecycline, fosfomycin, nitrofurantoin, chloramphenicol, rifampicin and trimethoprim/sulfamethoxazole, and by broth microdilution for colistin and interpreted according to EUCAST and CLSI breakpoints. WGS was performed on an Illumina MiSeq system. Contigs were obtained with the SPAdes genome assembler and genome annotation was conducted using RAST v.2.0 (http://rast.nmpdr.org/). Services provided by the Center for Genomic Epidemiology (https://cge.cbs.dtu.dk/services/) were used to identify resistance genes and the multilocus ST according to the Pasteur scheme (http://pubmlst.org/abaumannii). Geneious v.R8 (http://www.geneious.com) was used to analyse the genetic surroundings of resistance genes. The A. baumannii strain IHIT32296 was resistant to piperacillin, piperacillin/tazobactam, ceftiofur, cefpirome, meropenem, imipenem and nitrofurantoin, besides its intrinsic resistances to ampicillin, amoxicillin/clavulanic acid and first-generation cephalosporins. PCR typing2 revealed the presence of a variant of the blaOXA-24 gene, which was determined as blaOXA-72 by WGS. Using ResFinder 3.0 (https://cge.cbs.dtu.dk/services/ResFinder/), we additionally identified intrinsic β-lactamase genes ampC and blaOXA-343, which were not preceded by an insertion element. Whereas OXA-23 is widespread on all continents, the OXA-72 enzyme, which differs by only a single amino acid from OXA-24/40, has been mainly observed in A. baumannii isolates from South America, Southern Asia and Eastern Europe. However, its recent emergence in other countries, such as Mexico, France and Germany, suggests an ongoing spread of this oxacillinase type.5–7 Similar to OXA-23, which is still the most common acquired carbapenemase in A. baumannii, the dissemination and clonal spread of OXA-72 was often linked to strains of international clone (IC) 2 (sometimes referred to as a ‘worldwide’ or ‘global’ clone) and ST2 or related STs (Pasteur scheme).1,5 However, also non-IC2 strains have been reported for A. baumannii, such as ST78 in Germany and ST79, ST180 and ST730 in Brazil.5,7,8 MLST for our strain revealed ST294 (cpn60-40, fusA-3, gltA-2, pyrG-2, recA-4, rplB-35, rpoB-4). This ST has been identified for the first time in A. baumannii strains also carrying blaOXA-343, and was isolated from an artesian well and the mouth of a horse in Lebanon.9 The nucleotide sequence of the blaOXA-72-carrying plasmid pIHIT32296 was localized on a single contig. Its size of 8493 bp (Figure 1) was verified by PCR and subsequent sequence analysis of a 2550 bp amplicon to circularize the plasmid contig using primers repA-FP (5′-CACTGAAAGCTATCAACAGT-3′) and orf2-RP (5′-ATGGTATTACGCGTCAGCC-3′). The plasmid exhibited only partial similarity to different OXA-72-encoding plasmids that ranged between 8771 and 12 056 bp in size and were reported from various Acinetobacter species from human patients in different countries (Figure S1, available as Supplementary data at JAC Online).8,10–14 Plasmid pIHIT32296 encodes a replication protein (Rep) that belongs to the Rep 3 superfamily (pfam01051). It also carries genes for a putative type II toxin–antitoxin (TA) system (VapC2-VapB2) that are also present in other OXA-72-encoding plasmids, such as pAB-ML, pIEC338SCOX and pAP10253-1. Plasmid-encoded TA systems have been proposed to function as plasmid stabilization elements as they mediate post-segregational killing of daughter cells that have lost these plasmids. Different TA systems, such as RelB/RelE, HicA/HicB, SplT/SplA and MazE/F, have been found in clinical A. baumannii, but their role in the distribution and persistence of this pathogen or for the maintenance of resistance plasmids is not yet understood.15 The blaOXA-72 gene was flanked by XerC/XerD-binding sites. As previously suggested, Xer proteins may be involved in the mobilization of blaOXA-24 genes, as well as of other resistance genes, such as blaOXA-58, tet39 and msrE-mphE, within Acinetobacter plasmids and chromosomes by site-specific recombination mechanisms.16 Figure 1 View largeDownload slide Schematic representation of 8493 bp pIHIT32296, the blaOXA-72 gene-harbouring plasmid of A. baumannii IHIT32296. repA, replicase gene; ydaF2, gene encoding putative ribosomal N-acetyltransferase; vapB2, gene encoding putative antitoxin; vapC2, gene encoding putative ribonuclease/toxin; XerC/XerD, specific site of the tyrosine recombinases XerC and XerD (indicated by asterisks). Figure 1 View largeDownload slide Schematic representation of 8493 bp pIHIT32296, the blaOXA-72 gene-harbouring plasmid of A. baumannii IHIT32296. repA, replicase gene; ydaF2, gene encoding putative ribosomal N-acetyltransferase; vapB2, gene encoding putative antitoxin; vapC2, gene encoding putative ribonuclease/toxin; XerC/XerD, specific site of the tyrosine recombinases XerC and XerD (indicated by asterisks). In conclusion, we identified a carbapenem-resistant A. baumannii ST294 strain from a pet harbouring a unique blaOXA-72-carrying plasmid. The location of known genes encoding carbapenem resistance and TA systems on this plasmid demonstrates the possibility of plasmid maintenance. The acquisition of plasmids with beneficial properties by distinct A. baumannii strains may have resulted in the successful clonal lineages that emerged in hospitals worldwide, and their dissemination in other reservoirs (livestock/companion animals, environment) has only just begun. Nucleotide sequence accession number The nucleotide sequence of pIHIT32279-OXA-72 was deposited at GenBank under the accession number KY704308. The Whole Genome Shotgun project of strain IHIT32296 has been deposited at GenBank under the accession NWUK00000000. The version described in this paper is version NWUK01000000. Acknowledgements We thank the team of curators of the Institut Pasteur Acinetobacter MLST system for curating the data and making them publicly available at http://pubmlst.org/abaumannii/. Funding This work was supported by the Engemann Family Foundation and the Schaumann Foundation, providing grants to P. K., and by the Animal Health Academy, providing a grant to L. J. Transparency declarations None to declare. Supplementary data Figure S1 is available as Supplementary data at JAC Online. References 1 Higgins PG, Dammhayn C, Hackel M et al. Global spread of carbapenem-resistant Acinetobacter baumannii. J Antimicrob Chemother 2010; 65: 233– 8. 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Journal of Antimicrobial Chemotherapy – Oxford University Press
Published: Apr 1, 2018
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