title: Full characterization of the integrative and conjugative element carrying the metallo-β-lactamase bla SPM-1 and bicyclomycin bcr1 resistance genes found in the pandemic Pseudomonas aeruginosa clone SP/ST277.
abstract: OBJECTIVE
  This study aimed to characterize the genomic context of the bla SPM-1 gene in Brazilian strains belonging to the pandemic Pseudomonas aeruginosa clone SP/ST277.
  
  
  METHODS
  WGS of clone SP/ST277 strains was performed using a Nextera paired-end library in an Illumina HiSeq 2500 sequencer. bla SPM-1 context was assessed by de novo assembly and gene prediction and annotation tools. bla SPM-1 was screened in P. aeruginosa genomes through BlastN, and comparative genomics were performed.
  
  
  RESULTS
  The metallo-β-lactamase bla SPM-1 has been disseminated by the pandemic Brazilian P. aeruginosa clone SP/ST277. In spite of its association with the CR4 element and with the Tn4371 element, the overall bla SPM-1 genomic context remains uncharacterized and its determination is valuable to understanding gene dispersion dynamics and the consequent emergence of carbapenem resistance. In this study, bla SPM-1 and its surrounding sequences (CR4-groEL-bla SPM-1-CR4-groEL) were found in the variable region of an ICE-like element resembling Tn4371 (where ICE stands for integrative and conjugative element). This element, named ICETn4371 6061, had 46 ORFs, including the bicyclomycin resistance bcr1 gene. An integrase gene and a set of conjugative transfer genes were identified. Gene content and order were shared with other Tn4371-ICEs, presenting remarkable amino acid identities. bla SPM-1 and surrounding sequences were missing in ICETn4371 6061 of PS600-MA, another isolate belonging to clone SP/ST277, indicating their mobilization. Eight/nine P. aeruginosa genomes assigned to clone SP/ST277, by in silico MLST, harboured bla SPM-1 inserted into ICETn4371 6061.
  
  
  CONCLUSIONS
  The presence of bla SPM-1 in a Tn4371-ICE with intact integration/conjugation modules demonstrated that, besides gene dispersion by clonal expansion of the pandemic SP/ST277 lineage, bla SPM-1 may be spread through ICE conjugation.
authors.affiliation: Laboratório de Genética Molecular de Microrganismos, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brazil.
authors.forename: Erica L
authors.initials: EL
authors.lastname: Fonseca
authors.affiliation: Laboratório de Genética Molecular de Microrganismos, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brazil mfabanto@ioc.fiocruz.br.
authors.forename: Michel A
authors.initials: MA
authors.lastname: Marin
authors.affiliation: Laboratório de Genética Molecular de Microrganismos, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brazil.
authors.forename: Fernando
authors.initials: F
authors.lastname: Encinas
authors.affiliation: Laboratório de Genética Molecular de Microrganismos, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brazil.
authors.forename: Ana Carolina P
authors.initials: AC
authors.lastname: Vicente
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001938:Brazil / Q000453:epidemiology; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D003227:Conjugation, Genetic; D024881:Drug Resistance, Bacterial; D016680:Genome, Bacterial; D005838:Genotype; D020071:Interspersed Repetitive Sequences; D058889:Molecular Typing; D058873:Pandemics; D011552:Pseudomonas Infections / Q000453:epidemiology / Q000382:microbiology; D011550:Pseudomonas aeruginosa / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics; D017422:Sequence Analysis, DNA; D001618:beta-Lactamases / Q000235:genetics*; D047090:beta-Lactams / Q000494:pharmacology*
chemical_list: D000900:Anti-Bacterial Agents; D019086:Bridged Bicyclo Compounds, Heterocyclic; D047090:beta-Lactams; C476155:beta-lactamase SPM-1, Pseudomonas aeruginosa; D001618:beta-Lactamases; C024504:bicozamycin