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6 Department of Epidemiology and Clinical Microbiology, National Medicines Institute, 00-
9 Poland
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10 Microbiological Laboratory, University Hospital of Lord’s Transfiguration, 61-848 Poznań,
11 Poland
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13 *Corresponding author. Tel: +48 22 851 46 70; Fax: +48 22 841 29 49; E-mail:
14 e.literacka@nil.gov.pl
16 OXA-58
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18 OXA-58 represents one of the class D carbapenemase types that are major carbapenem
20 Its first report in diverse enterobacteria was from Sierra Leone (4), followed by recent papers
21 on Proteus mirabilis from Belgium (5) and Germany (6), the latter one describing specific
23 strains. Here we present a Polish isolate with an almost identical plasmid, raising questions on
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25 Carbapenem-resistant, Carba NP test-positive (7) P. mirabilis NMI1213/17 was recovered in
27 OXA-58 was indicated by the eazyplex SuperBug CRE assay (AmplexDiagnostics, Giessen,
28 Germany), and confirmed by PCR and sequencing. Antimicrobial MICs were evaluated by
35 fosfomycin.
36 The isolate was subjected to whole-genome sequencing (WGS), using the MiSeq platform
37 (Illumina, San Diego, USA). Contigs were assembled and the draft genome obtained using
38 SPAdes 3.12.0 (8), annotation was done with PROKKA 1.11 (9), and resistance genes were
39 identified by ResFinder 3.0 (10). blaOXA-58 was on a 6,351bp-long contig with identical
40 termini, suggesting a circular DNA molecule, confirmed by PCR and Sanger sequencing. The
41 entire plasmid pPOZ-OXA-58 was of 6,224bp (GenBank accession No. MK086028), and
42 when compared with p10797-OXA-58 (6,219bp) from Germany (KU871396) (6), it was
43 almost identical, with two putative rep genes, repA and rep, blaOXA-58 flanked by remnants of
45 gene (6). The only differences were insertion of five adenine nucleotides in rep in pPOZ-
46 OXA-58, and one single-nucleotide polymorphism in the ISAba3 tnpA gene. As revealed by
47 Lange et al. (6), the rep and aadA14 genes are present also in the 5,198-bp plasmid pCCK647
48 (AJ884726) from Pasteurella multocida from Belgium (11). However, the only pCCK647
49 replicase gene rep is presumably non-active in pPOZ-OXA-58 due to the (A)5 frameshift
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50 insertion; therefore, its replication might rely on repA, and proceed in a broader host range, as
52 (Invitrogen, Carlsbad, CA). P. mirabilis NMI1213/17 contained only one more acquired
54 several P. mirabilis WGS entries (e.g. CP015347). The gyrA and parC genes specified DNA
57 PFGE to any of the four German P. mirabilis OXA-58 strains (obtained by courtesy of N.
60 marginal, though puzzling epidemiological story. Only six such isolates, exclusively P.
61 mirabilis, have been reported in Europe so far, of which German and Polish isolates shared
62 unique plasmids with the truncated blaOXA-58-carrying ISAba3-associated transposon (3). The
63 plasmids might spread horizontally by mobilised transfer (6), and occurrence of the isolates in
64 distant locales might suggest hidden dissemination of unknown scale and future potential.
65 Accession number. The nucleotide sequence obtained in this study has been submitted to the
67 Acknowledgments
68 This work was supported by the grant SPUB MIKROBANK from the Polish Ministry of
69 Science and Higher Education, Narodowy Program Ochrony Antybiotyków from the Polish
70 Ministry of Health, and grant DS-4.47/2018 from the National Medicines Institute. All of the
72 References
73 1. Evans BA, Amyes SG. 2014. OXA β-lactamases. Clin Microbiol Rev 27:241-263.
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74 2. Poirel L, Naas T, Nordmann P. 2010. Diversity, epidemiology, and genetics of class D
79 4. Leski TA, Bangura U, Jimmy DH, Ansumana R, Lizewski SE, Li RW, Stenger DA,
86 Chemother 61:e01697-16.
92 8. Bankevich A, Nurk S, Antipov D, Gurevich AA, Dvorkin M, Kulikov AS, Lesin VM,
93 Nikolenko SI, Pham S, Prjibelski AD, Pyshkin AV, Sirotkin AV, Vyahhi N, Tesler G,
94 Alekseyev MA, Pevzner PA. 2012. SPAdes: a new genome assembly algorithm and
97 30:2068-2069.
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98 10. Zankari E, Hasman H, Cosentino S, Vestergaard M, Rasmussen S, Lund O, Aarestrup
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105 Figure legends
106 Figure 1. Scheme of the pPOZ-OXA-58 plasmid structure (6,224bp; GenBank acc. No.
107 MK086028). Black arrows represent genes or putative genes, and black boxes represent the
108 IR-L and IR-R sequences of two truncated ISAba3 elements, respectively. Two differences
109 with the plasmid p10797-OXA-58 (KU871396) are indicated above the scheme, namely the
110 (A)5 insertion into the original (A)6 tract at the position 174-179 of the rep coding region, and
112 individual arrows/boxes and their positions only roughly correspond to the actual plasmid
113 structure.
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