title: Extended-Spectrum β-Lactamase-Producing Klebsiella pneumoniae from Pharmaceutical Wastewaters in South-Western Nigeria.
abstract: Emergence and spread of Klebsiella pneumoniae isolates producing extended-spectrum β-lactamases (ESBLs) present a major threat to public health. In this study, we characterized β-lactam-resistant K. pneumoniae isolates from six wastewater samples obtained from two pharmaceutical industries located in Lagos and Ogun States, Nigeria. Bacteria were isolated by using MacConkey agar; species identification and antibacterial susceptibility testing were performed by Vitek 2. Etest was used for ESBL phenotype confirmation. The presence of β-lactamase genes was investigated by PCR and sequencing. Bacterial strain typing was done by XbaI-macrorestriction and subsequent pulsed-field gel electrophoresis (PFGE) as well as multilocus sequence typing (MLST). Thirty-five bacterial species were isolated from the six samples; among them, we identified seven K. pneumoniae isolates with resistance to β-lactams and co-resistance to fluoroquinolones, aminoglycosides, and folate pathway inhibitors. The ESBL phenotype was confirmed in six K. pneumoniae isolates that harbored ESBL genes blaCTX-M-15(n = 5), blaSHV-2(n = 1), and blaSHV-12(n = 1). PFGE and MLST analysis revealed five clones belonging to four sequence types (ST11, ST15, ST37, ST101), and clone K. pneumoniae-ST101 was present in the wastewater samples from two different pharmaceutical industries. Additionally performed conjugation assays confirmed the location of β-lactamase genes on conjugative plasmids. This is the first confirmation of K. pneumoniae isolates producing CTX-M-15-ESBL from pharmaceutical wastewaters in Nigeria. The co-resistance observed might be a reflection of the different drugs produced by these industries. Continuous surveillance of the environmental reservoirs of multidrug-resistant bacteria is necessary to prevent their further spread.
authors.affiliation: 1 Department of Microbiology, University of Lagos , Lagos, Nigeria .
authors.forename: Avemaria
authors.initials: A
authors.lastname: Obasi
authors.affiliation: 1 Department of Microbiology, University of Lagos , Lagos, Nigeria .
authors.forename: SimonCyril
authors.initials: S
authors.lastname: Nwachukwu
authors.affiliation: 1 Department of Microbiology, University of Lagos , Lagos, Nigeria .
authors.forename: Esther
authors.initials: E
authors.lastname: Ugoji
authors.affiliation: 2 Department of Medical Microbiology, Friedrich Loeffler Institute for Medical Microbiology, University of Greifswald , Greifswald, Germany .
authors.forename: Christian
authors.initials: C
authors.lastname: Kohler
authors.affiliation: 2 Department of Medical Microbiology, Friedrich Loeffler Institute for Medical Microbiology, University of Greifswald , Greifswald, Germany .
authors.forename: André
authors.initials: A
authors.lastname: Göhler
authors.affiliation: 2 Department of Medical Microbiology, Friedrich Loeffler Institute for Medical Microbiology, University of Greifswald , Greifswald, Germany .
authors.forename: Veronica
authors.initials: V
authors.lastname: Balau
authors.affiliation: 3 FG13 Nosocomial Pathogens and Antibiotic Resistance, Robert Koch Institute , Wernigerode, Germany .
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Pfeifer
authors.affiliation: 2 Department of Medical Microbiology, Friedrich Loeffler Institute for Medical Microbiology, University of Greifswald , Greifswald, Germany .
authors.forename: Ivo
authors.initials: I
authors.lastname: Steinmetz
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000378:metabolism; D015373:Bacterial Typing Techniques / Q000379:methods; D004345:Drug Industry / Q000379:methods; D024901:Drug Resistance, Multiple, Bacterial / Q000187:drug effects; D007711:Klebsiella pneumoniae / Q000187:drug effects / Q000378:metabolism*; D017720:Molecular Epidemiology / Q000379:methods; D009549:Nigeria; D062065:Wastewater / Q000382:microbiology*; D001618:beta-Lactamases / Q000378:metabolism*; D047090:beta-Lactams / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D062065:Wastewater; D001618:beta-Lactamases; D047090:beta-Lactams
keywords: CTX-M-15; SHV-12; SHV-2; conjugative plasmids; multidrug resistance