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mSphere
2019 Oct 16;45:. doi: 10.1128/mSphere.00376-19.
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Genetic Characterization of Plasmid-Borne blaOXA-58 in Distinct Acinetobacter Species.
Matos AP
,
Cayô R
,
Almeida LGP
,
Streling AP
,
Nodari CS
,
Martins WMBS
,
Narciso AC
,
Silva RM
,
Vasconcelos ATR
,
Gales AC
.
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We characterize by whole-plasmid-sequence (WPS) two-plasmid-borne blaOXA-58 obtained from Acinetobacter seifertii (Asp-1069) and A. baumannii (Acb-45063) clinical strains recovered 17 years apart from distinct Brazilian regions. Multilocus sequence type (MLST) analysis showed that the Asp-1069 and Acb-45063 strains belong to ST551 and ST15/CC15, respectively. WPS analysis demonstrated that blaOXA-58 was located in two distinct plasmids named pAs1069_a (24,672 bp/44 open reading frames [ORFs]) and pAb45063_b (19,808 bp/24 ORFs), which belong to the GR8/GR23 (repAci23) and GR4 (repAci4) incompatibility groups, respectively. The genetic environments surrounding blaOXA-58 revealed that it was flanked by two intact ISAba3 copies on pAb45063_b, which differed from pAs1069_a. In the latter, the upstream ISAba3 copy was truncated by insertion of ISAba825 element. Although Re27-specific recombination sites were found adjacent to ISAba3-blaOXA-58-ISAba3 arrangement on pAb45063_b, such structures were absent on pAs1069_a. The conserved ISAba125-araC1-lysE arrangement was disrupted by TnaphA6 harboring the aminoglycosides resistance gene aphA6 on pAs1069_a, while an IS26-blaTEM-1-aac(3)-IIa-IS26 genetic structure was found upstream from ISAba3-blaOXA-58-ISAba3 on pAb45063_b. Other two plasmids, pAb45063_a (183,767 bp/209 ORFs) and pAs1069_b (13,129 bp/14 ORFs), were also found in the OXA-58-producing Acinetobacter species strains, harboring the strA and strB genes and the sul2 gene, which confer resistance to streptomycin and sulfonamides, respectively. The plasmid-mediated virulence factors corresponding to genes tonB, spl, glmM, ppa, sulP, and map were found in both strains, as well distinct toxin-antitoxin system-encoding genes stbD and relE (pAs1069_a), brnT and brnA (pAb45063_b), and xreE (pAb45063_a). Although infrequently reported in Brazil, plasmid-borne blaOXA-58 showed a complex and diverse genetic backbone that confers stability in different Acinetobacter species that have been isolated from nosocomial settings over time.IMPORTANCE Although the blaOXA-58 gene has been infrequently described in Brazil, contrasting with other bordering South American countries, we verified the maintenance of this resistance determinant over time among carbapenem-resistant Acinetobacter species isolates, not only in nosocomial settings but also in the environment. In addition, to the best of our knowledge, this is the first study to have used WPS analysis to evaluate the genetic surroundings of blaOXA-58 in Brazil. Moreover, the A. seifertii and A. baumannii clinical strains evaluated in this study were recovered 17 years apart in hospitals located in distinct Brazilian geographic regions.
FIG 1. Schematic representation of circular plasmid maps found in A. seifertii Asp-1069 (A and C) and A. baumannii Acb-45063 (B and D) strains. Arrows designate transcription directions of genes and ORFs. Genes were grouped according to their predictive functions as indicated by the color coding key.
FIG 2. Genetic contexts surrounding blaOXA-58 found in plasmids pAb45063_b (A) and pAs1069_a (B). Genes and their transcriptional orientations are represented by horizontal arrows. Identical genes found in both genetic structures are represented with the same colors. Genes of no predicted functions (HP [hypothetical proteins]) are represented in white. The putative original promoters driving the expression of blaOXA-58 genes are highlighted. Direct repeat sequences are represented by an asterisk (*). Gene names preceded by an uppercase Greek delta (Δ) represent truncated genes, and the corresponding regions are shaded in gray. The Re27 regions are boxed. Promoter prediction for blaOXA-58 was performed using BPROM (SoftBerry).
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