Abstract

BackgroundWe have recovered one blaNDM-1-harboring bacterial strain, designated as XM1570, from a sputum sample obtained from a fatal case of pneumonia in China.MethodsBiochemical profiling, 16S rRNA sequencing and antimicrobial susceptibility testing were performed. Conjugation experiments were conducted to determine transmissibility of resistance. Pulsed-field gel electrophoresis and whole genome sequencing were performed to identify strain-specific features.ResultsThe isolate XM1570 was identified as Acinetobacter calcoaceticus. Whole genome sequencing identified two plasmids, pXM1 and pXM2. Comparative analysis showed >99% similarity between XM1570 and A. calcoaceticus PHEA-2. Plasmid pXM1 carried the carbapenemase gene blaNDM-1 and displayed high homology with previously described plasmids isolated from different Acinetobacter spp., which were collected from human or livestock distributed in China and worldwide. The blaNDM-1 gene was located on this conjugative plasmid in a transposon-like region flanked by two copies of the insertion sequence ISAba125; and resistance to all tested β-lactams was observed. Transferability of resistance from pXM1 to the transconjugants was identified. Plasmid pXM2 had an insertion sequence ISAba125 and a −35 region of the blaNDM-1 gene promoter but the blaNDM-1 gene was not present. A chromosomally located carbapenemase-encoding gene blaOXA-75 was detected; however, this gene was interrupted by an insertion sequence ISAba22 belonging to IS3 family.ConclusionsLocation of blaNDM-1 on different self-transmissible plasmids could facilitate geographically broad dissemination and host range expansion of the blaNDM-1 gene via horizontal gene transfer. Our findings of this normally environmental species A. calcoaceticus XM1570 further underline the significant clinical challenge and the essential need for surveillance including molecular methods and plasmid analyses.Electronic supplementary materialThe online version of this article (doi:10.1186/s12879-015-0870-7) contains supplementary material, which is available to authorized users.

Highlights

  • We have recovered one blaNDM-1-harboring bacterial strain, designated as XM1570, from a sputum sample obtained from a fatal case of pneumonia in China

  • Of concern was the reported identification of 27 Acinetobacter spp. isolates with the blaNDM-1 gene recovered from intensive care units in China [15]

  • The isolate was positive for the blaNDM-1 gene and identified as A. calcoaceticus, which is a rare human pathogen but a species frequently recovered from soil and water with no implication in serious human diseases [25]

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Summary

Methods

Case report and bacterial isolates During routine sentinel surveillance, we isolated a blaNDM-1-harboring bacterial strain from a patient died of pneumonia and respiratory failure. A. calcoaceticus XM1570 was isolated from the patient described above. Other 22 Acinetobacter spp. isolates were recovered from sputum samples of patients in the same hospital during May-July 2010. Transconjugants, selected for by growth on ampicillin, were confirmed by 16S rRNA sequencing and transferability of the blaNDM-1 gene was verified by PCR and sequencing. Nih.gov/genomes/Bacteria/) and compared with those of A. calcoaceticus XM1570 to generate core conserved proteins, which were defined with a minimum of 95% identity and coverage. Those conserved proteins were concatenated and aligned by ProbCons with default options [31]. Sequences of plasmids with high homology were downloaded from NCBI, such as p3ABAYE [GenBank: CU459140], pMS32-1 [GenBank: KJ616405], pNDM-BJ01 [GenBank: JQ001791], pNDM-BJ02 [GenBank: JQ060896], and pNDM-AB [GenBank: KC503911]

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