Acinetobacter calcoaceticus from a fatal case of pneumonia harboring blaNDM-1 on a widely distributed plasmid

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.

[1]  A. Carattoli,et al.  Distribution of Intrinsic Plasmid Replicase Genes and Their Association with Carbapenem-Hydrolyzing Class D β-Lactamase Genes in European Clinical Isolates of Acinetobacter baumannii , 2011, Antimicrobial Agents and Chemotherapy.

[2]  D. van Duin,et al.  Carbapenem-resistant Enterobacteriaceae: A menace to our most vulnerable patients , 2013, Cleveland Clinic Journal of Medicine.

[3]  Yunsong Yu,et al.  Epidemiological characteristics and genetic structure of blaNDM-1 in non-baumannii Acinetobacter spp. in China. , 2012, The Journal of antimicrobial chemotherapy.

[4]  G. Cornaglia,et al.  New Delhi metallo-beta-lactamase (NDM-1): towards a new pandemia? , 2010, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[5]  Yunsong Yu,et al.  Emergence of NDM-1-producing Acinetobacter baumannii in China. , 2011, The Journal of antimicrobial chemotherapy.

[6]  D. Livermore,et al.  Dissemination of NDM-1 positive bacteria in the New Delhi environment and its implications for human health: an environmental point prevalence study. , 2011, The Lancet. Infectious diseases.

[7]  N. Perna,et al.  progressiveMauve: Multiple Genome Alignment with Gene Gain, Loss and Rearrangement , 2010, PloS one.

[8]  Ruifu Yang,et al.  Novel Plasmid and Its Variant Harboring both a blaNDM-1 Gene and Type IV Secretion System in Clinical Isolates of Acinetobacter lwoffii , 2012, Antimicrobial Agents and Chemotherapy.

[9]  Chuong B. Do,et al.  ProbCons: Probabilistic consistency-based multiple sequence alignment. , 2005, Genome research.

[10]  D. Ecker,et al.  Analysis of Antibiotic Resistance Genes in Multidrug-Resistant Acinetobacter sp. Isolates from Military and Civilian Patients Treated at the Walter Reed Army Medical Center , 2006, Antimicrobial Agents and Chemotherapy.

[11]  Timothy R. Walsh,et al.  Characterization of a New Metallo-β-Lactamase Gene, blaNDM-1, and a Novel Erythromycin Esterase Gene Carried on a Unique Genetic Structure in Klebsiella pneumoniae Sequence Type 14 from India , 2009, Antimicrobial Agents and Chemotherapy.

[12]  Haiying Yu,et al.  Genome Sequence of Acinetobacter calcoaceticus PHEA-2, Isolated from Industry Wastewater , 2011, Journal of bacteriology.

[13]  Thierry Naas,et al.  Global Spread of Carbapenemase-producing Enterobacteriaceae , 2011, Emerging infectious diseases.

[14]  L. Frost,et al.  F factor conjugation is a true type IV secretion system. , 2003, FEMS microbiology letters.

[15]  P. Nordmann,et al.  Genetic and Biochemical Characterization of the First Extended-Spectrum CARB-Type ß-Lactamase, RTG-4, from Acinetobacter baumannii , 2009, Antimicrobial Agents and Chemotherapy.

[16]  Paul Keim,et al.  Evolution of a Pathogen: A Comparative Genomics Analysis Identifies a Genetic Pathway to Pathogenesis in Acinetobacter , 2013, PloS one.

[17]  Detection of New Delhi Metallo-β-Lactamase (Encoded by bla NDM-1) in Acinetobacter schindleri during Routine Surveillance , 2013, Journal of Clinical Microbiology.

[18]  Robert J. Clifford,et al.  Bacterial Peritonitis Due to Acinetobacter baumannii Sequence Type 25 with Plasmid-Borne New Delhi Metallo-β-Lactamase in Honduras , 2013, Antimicrobial Agents and Chemotherapy.

[19]  Siguo Liu,et al.  Complete sequence of the bla(NDM-1)-carrying plasmid pNDM-AB from Acinetobacter baumannii of food animal origin. , 2013, The Journal of antimicrobial chemotherapy.

[20]  J. Iredell,et al.  Genetic Contexts of blaNDM-1 , 2012, Antimicrobial Agents and Chemotherapy.

[21]  Liangxing Wang,et al.  Emergence of blaNDM-1 among Klebsiella pneumoniae ST15 and novel ST1031 clinical isolates in China. , 2013, Diagnostic microbiology and infectious disease.

[22]  C. Chao,et al.  Acinetobacter calcoaceticus–Acinetobacter baumannii Complex Is Not Equal to A. baumannii , 2013, Infection Control & Hospital Epidemiology.

[23]  Liuyu Huang,et al.  Higher Isolation of NDM-1 Producing Acinetobacter baumannii from the Sewage of the Hospitals in Beijing , 2013, PloS one.

[24]  Yufei Wang,et al.  Coexistence of blaNDM-1 with the prevalent blaOXA23 and blaIMP in pan-drug resistant Acinetobacter baumannii isolates in China. , 2011, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[25]  K. Peck,et al.  Epidemiology and Clinical Features of Community-Onset Acinetobacter baumannii Infections , 2012, Infection Control & Hospital Epidemiology.

[26]  Qijing Zhang,et al.  Identification of New Delhi Metallo-β-lactamase 1 in Acinetobacter lwoffii of Food Animal Origin , 2012, PloS one.

[27]  J. Poulain,et al.  Comparative Analysis of Acinetobacters: Three Genomes for Three Lifestyles , 2008, PloS one.

[28]  Yufei Wang,et al.  High rate of New Delhi metallo-β-lactamase 1-producing bacterial infection in China. , 2013, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[29]  Weijun Chen,et al.  Complete Genome Sequence of an Acinetobacter Strain Harboring the NDM-1 Gene , 2013, Genome Announcements.

[30]  P. Courvalin,et al.  Loss of intrinsic aminoglycoside resistance in Acinetobacter haemolyticus as a result of three distinct types of alterations in the aac(6')-Ig gene, including insertion of IS17 , 1997, Antimicrobial agents and chemotherapy.

[31]  Y. Zhou,et al.  Dissemination and characterization of NDM-1-producing Acinetobacter pittii in an intensive care unit in China. , 2012, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[32]  N. Woodford,et al.  Carbapenemase-producing Enterobacteriaceae and non-Enterobacteriaceae from animals and the environment: an emerging public health risk of our own making? , 2014, The Journal of antimicrobial chemotherapy.

[33]  J. Vila,et al.  A comparative study of different PCR-based DNA fingerprinting techniques for typing of the Acinetobacter calcoaceticus-A. baumannii complex. , 1996, Journal of medical microbiology.

[34]  Harald Seifert,et al.  Acinetobacter baumannii: Emergence of a Successful Pathogen , 2008, Clinical Microbiology Reviews.

[35]  V. Cheng,et al.  Emergence of NDM-1-producing Enterobacteriaceae in China. , 2012, The Journal of antimicrobial chemotherapy.

[36]  O. Gascuel,et al.  New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. , 2010, Systematic biology.

[37]  Z. Zong,et al.  blaNDM-1-carrying Acinetobacter johnsonii detected in hospital sewage. , 2013, The Journal of antimicrobial chemotherapy.

[38]  Göran Tomson,et al.  Antibiotic resistance in China—a major future challenge , 2009, The Lancet.

[39]  J. Schrenzel,et al.  Tn125-Related Acquisition of blaNDM-Like Genes in Acinetobacter baumannii , 2011, Antimicrobial Agents and Chemotherapy.

[40]  Robert A. Bonomo,et al.  Carbapenems: Past, Present, and Future , 2011, Antimicrobial Agents and Chemotherapy.

[41]  Wei Li,et al.  Identification and Characterization of the First Escherichia coli Strain Carrying NDM-1 Gene in China , 2013, PloS one.

[42]  J. Charan,et al.  New Delhi Metallo – beta lactamase – 1 containing Enterobacteriaceae: Origin, Diagnosis, Treatment and Public health concern , 2012, The Pan African medical journal.

[43]  M. Ferraro Performance standards for antimicrobial susceptibility testing , 2001 .