Autor(es):
Mesquita, Cristina S ; Soares-Castro, Pedro ; Faustino, Alberta ; Santos, Hugo M ; Capelo, José L ; Santos, P. M.
Data: 2019
Identificador Persistente: https://hdl.handle.net/1822/73222
Origem: RepositóriUM - Universidade do Minho
Assunto(s): Adolescent; Adult; Aged; Aged, 80 and over; Child; Child, Preschool; Cluster Analysis; Female; Hospitals; Humans; Infant; Infant, Newborn; Male; Middle Aged; Molecular Typing; Pneumonia, Bacterial; Portugal; Pseudomonas Infections; Pseudomonas aeruginosa; Random Amplified Polymorphic DNA Technique; Virulence Factors; Young Adult; Biological Variation, Population; Genetic Loci; Genotype; epidemiological study; Genotype profiling; Antimicrobial resistance; Comparative genomics; Science & Technology; Ciências Naturais::Ciências Biológicas
Descrição
In this work, a genotype-phenotype survey of a highly diversified Pseudomonas aeruginosa collection was conducted, aiming to detail pathogen-associated scenarios that clinicians face nowadays. Genetic relation based on RAPD-PCR of 705 isolates, retrieved from 424 patients and several clinical contexts, reported an almost isolate-specific molecular-pattern. Pneumonia-associated isolates HB13 and HB15, clustered in the same RAPD-PCR group, were selected to evaluate the genomic background underlying their contrasting antibiotic resistance and virulence. The HB13 genome harbors antibiotic-inactivating enzymes-coding genes (e.g. aac(3)-Ia, arr, blaVIM-2) and single-nucleotide variations (SNVs) in antibiotic targets, likely accounting for its pan-resistance, whereas HB15 susceptibility correlated to predicted dysfunctional alleles. Isolate HB13 showed the unprecedented rhl-cluster absence and variations in other pathogen competitiveness contributors. Conversely, HB15 genome comprises exoenzyme-coding genes and SNVs linked to increased virulence. Secretome analysis identified signatures features with unknown function as potential novel pathogenic (e.g. a MATE-protein in HB13, a protease in HB15) and antibiotic resistance (a HlyD-like secretion protein in HB13) determinants. Detection of active prophages, proteases (including protease IV and alkaline metalloproteinase), a porin and a peptidase in HB15 highlights the secreted arsenal likely essential for its virulent behavior. The presented phenotype-genome association will contribute to the current knowledge on Pseudomonas aeruginosa pathogenomics.
This work was supported by the strategic programme UID/BIA/0050/2013 (POCI-01-0145-FEDER-007569) funded by national funds through FCT I.P., by ERDF through the COMPETE2020 - Programa Operacional Competitividade e Internacionalização (POCI) and through a PhD grant (SFRH/BD/98558/2013) attributed to C.S.M. The facility for Biological Mass Spectrometry Isabel Moura was funded by Proteomass Scientific Society. H.M.S. is funded by the FCT 2015 Investigator Program (IF/00007/2015).
