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1.
PLoS One ; 16(7): e0254805, 2021.
Article in English | MEDLINE | ID: mdl-34310625

ABSTRACT

The purpose of the study was to characterize the resistome, virulome, mobilome and Clustered Regularly Interspaced Short Palindromic Repeats-associated (CRISPR-Cas) system of extended-spectrum ß-lactamase producing Klebsiella pneumoniae (ESBL-KP) clinical isolates and to determine their phylogenetic relatedness. The isolates were from Algeria, isolated at the University Hospital Establishment of Oran, between 2011 and 2012. ESBL-KP isolates (n = 193) were screened for several antibiotic resistance genes (ARGs) using qPCR followed by Pulsed-Field Gel Electrophoresis (PFGE). Representative isolates were selected from PFGE clusters and subjected to whole-genome sequencing (WGS). Genomic characterization of the WGS data by studying prophages, CRISPR-Cas systems, Multi-Locus Sequence Typing (MLST), serotype, ARGs, virulence genes, plasmid replicons, and their pMLST. Phylogenetic and comparative genomic were done using core genome MLST and SNP-Based analysis. Generally, the ESBL-KP isolates were polyclonal. The whole genome sequences of nineteen isolates were taken of main PFGE clusters. Sixteen sequence types (ST) were found including high-risk clones ST14, ST23, ST37, and ST147. Serotypes K1 (n = 1), K2 (n = 2), K3 (n = 1), K31 (n = 1), K62 (n = 1), and K151 (n = 1) are associated with hyper-virulence. CRISPR-Cas system was found in 47.4%, typed I-E and I-E*. About ARGs, from 193 ESBL-KP, the majority of strains were multidrug-resistant, the CTX-M-1 enzyme was predominant (99%) and the prevalence of plasmid-mediated quinolone resistance (PMQR) genes was high with aac(6')-lb-cr (72.5%) and qnr's (65.8%). From 19 sequenced isolates we identified ESBL, AmpC, and carbapenemase genes: blaCTX-M-15 (n = 19), blaOXA-48 (n = 1), blaCMY-2 (n = 2), and blaCMY-16 (n = 2), as well as non-ESBL genes: qnrB1 (n = 12), qnrS1 (n = 1) and armA (n = 2). We found IncF, IncN, IncL/M, IncA/C2, and Col replicon types, at least once per isolate. This study is the first to report qnrS in ESBL-KP in Algeria. Our analysis shows the concerning co-existence of virulence and resistance genes and would support that genomic surveillance should be a high priority in the hospital environment.


Subject(s)
Bacterial Proteins/genetics , Klebsiella Infections/drug therapy , Klebsiella pneumoniae/enzymology , beta-Lactamases/genetics , Algeria , Anti-Bacterial Agents/adverse effects , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/metabolism , Clone Cells/cytology , Clone Cells/drug effects , Drug Resistance, Multiple, Bacterial/drug effects , Drug Resistance, Multiple, Bacterial/genetics , Genome, Bacterial/genetics , Hospitals, University , Humans , Klebsiella Infections/enzymology , Klebsiella Infections/microbiology , Klebsiella Infections/pathology , Klebsiella pneumoniae/pathogenicity , Microbial Sensitivity Tests , Multilocus Sequence Typing , Phylogeny , Plasmids/drug effects , Whole Genome Sequencing , beta-Lactamases/metabolism
2.
Microb Drug Resist ; 26(12): 1437-1441, 2020 Dec.
Article in English | MEDLINE | ID: mdl-31829797

ABSTRACT

Objectives: Pseudomonas aeruginosa occupies a central position in nosocomial infections and remains a significant cause of morbidity and mortality. The aim of this study was to characterize carbapenem resistance mechanisms in P. aeruginosa isolates from clinical specimens collected at the University Hospital of Oran, western Algeria. Materials and Methods: The identification of 214 nonduplicated P. aeruginosa isolates (collected from January to December 2016) was confirmed using matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Thirteen antibiotics were tested using the disc diffusion method. Carbapenemase-encoding genes were detected with the GeneXpert system and multiplex polymerase chain reaction (PCR). Clonal relatedness was determined using multilocus sequence typing (MLST) and the seven housekeeping genes were further used for phylogenetic analysis of imipenem-resistant P. aeruginosa using concatenated gene fragments. The flanking regions of the blaVIM-4 gene were analyzed by whole-genome sequencing. Results: Eleven isolates (5.39%) were resistant to carbapenems. PCR amplification and sequencing showed that six of these isolates (2.94%) harbored the blaVIM-4 gene that was carried on a novel class 1 integron. MLST analysis assigned the tested isolates to seven different sequence types (STs), of which two were new (ST3349 and ST3350) and five were previously described (ST244, ST499, ST709, ST809, and ST1239). Conclusion: In this study, we reported P. aeruginosa isolates producing VIM-4 in an Algerian hospital. The blaVIM-4 is harbored in class 1 integron with a new arrangement of genes cassettes.


Subject(s)
Carbapenems/pharmacology , Pseudomonas aeruginosa/drug effects , Pseudomonas aeruginosa/genetics , beta-Lactamases/genetics , Adult , Algeria , Bacterial Proteins/genetics , Bacteriological Techniques , Drug Resistance, Bacterial/genetics , Female , Humans , Integrons/genetics , Male , Middle Aged , Multilocus Sequence Typing , Polymerase Chain Reaction , Pseudomonas aeruginosa/isolation & purification
4.
Emerg Infect Dis ; 13(10): 1459-62, 2007 Oct.
Article in English | MEDLINE | ID: mdl-18257987

ABSTRACT

An outbreak of plague occurred in the region of Oran, Algeria, from June to July 2003. Algeria had not reported this disease for >50 years. Eighteen bubonic cases were identified, and Yersinia pestis was isolated from 6 patients. Except for the index case-patient, all patients recovered. Targeted chemoprophylaxis, sanitation, and vector control played a crucial role in controlling the outbreak. Epidemiologic and biomolecular findings strongly suggested the existence of a local animal reservoir during this period, but its origin (resurgence or re-importation) could not be determined. This sudden and unexpected reemergence of plague, close to an important commercial seaport, is a textbook illustration of a public health event of international importance. It also demonstrates that the danger of plague reoccurrence is not limited to the currently indexed natural foci.


Subject(s)
Contact Tracing , Disease Outbreaks/prevention & control , Plague/epidemiology , Algeria/epidemiology , Animals , Antibiotic Prophylaxis/methods , Disease Reservoirs , Disease Vectors , Humans , Rodent Diseases/microbiology , Rodent Diseases/transmission , Sanitation , Siphonaptera/microbiology
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