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  • 1.
    Goswami, Manish
    et al.
    School of Science and Technology, Örebro University, Örebro, Sweden; Molecular Biology Division Bhabha Atomic Research Centre, Mumbai, India; Homi Bhabha National Institute, Anushaktinagar, Mumbai, India.
    Khan, Faisal Ahmad
    Örebro University, School of Science and Technology.
    Ibrisevic, Admir
    Eskilstuna Strängnäs Energi & Miljö AB, Eskilstuna, Sweden.
    Olsson, Per-Erik
    Örebro University, School of Science and Technology.
    Jass, Jana
    Örebro University, School of Science and Technology.
    Development of Escherichia coli-based gene expression profiling of sewage sludge leachates2018In: Journal of Applied Microbiology, ISSN 1364-5072, E-ISSN 1365-2672, Vol. 125, no 5, p. 1502-1517Article in journal (Refereed)
    Abstract [en]

    AIMS: The impact of municipal waste on pathogenic microorganisms released into the environment is a public health concern. The present study aims to evaluate the effects of sewage sludge and antibiotic contaminants on stress response, virulence and antibiotic resistance in a pathogenic Escherichia coli.

    METHODS AND RESULTS: The effects of sewage sludge leachates on uropathogenic E. coli CFT073 were determined by monitoring the expression of 45 genes associated with antibiotic/metal resistance, stress response and virulence using RT-qPCR. The E. coli gene expression was validated using sub-inhibitory concentrations of tetracycline and ciprofloxacin. E. coli exposed to sewage sludge or sewage sludge-fly ash leachates altered the expression of 5 antibiotic and metal resistance, 3 stress response and 2 virulence associated genes. When antibiotics were combined with sludge or sludge-fly ash the antibiotic-associated gene expression was altered.

    CONCLUSIONS: E. coli treated with two sludge leachates had distinct gene expression patterns that were altered when the sludge leachates were combined with tetracycline, although to a lesser extent with ciprofloxacin.

    SIGNIFICANCE AND IMPACT OF STUDY: The E. coli multigene expression analysis is a potential new tool for assessing the effects of pollutants on pathogenic microbes in environmental waters for improved risk assessment.

  • 2.
    Khan, Faisal Ahmad
    et al.
    Örebro University, School of Science and Technology.
    Hellmark, Bengt
    Örebro University, School of Health Sciences. Örebro University Hospital.
    Ehricht, Ralf
    Abbott (Alere Technologies GmbH), Jena, Germany; InfectoGnostics Research Campus, Jena, Germany; Research Alliance - Leibniz Health Technologies, Leibniz Institute of Photonic Technology, Jena, Germany.
    Söderquist, Bo
    Örebro University, School of Medical Sciences.
    Jass, Jana
    Örebro University, School of Science and Technology.
    Related carbapenemase-producing Klebsiella isolates detected in both a hospital and associated aquatic environment in Sweden2018In: European Journal of Clinical Microbiology and Infectious Diseases, ISSN 0934-9723, E-ISSN 1435-4373, Vol. 37, no 12, p. 2241-2251Article in journal (Refereed)
    Abstract [en]

    Carbapenem antibiotics are one of the last-resort agents against multidrug-resistant (MDR) bacteria. The occurrence of carbapenemase-producing Enterobacteriaceae (CPE) in wastewater and aquatic environments is an indication of MDR bacteria in the community. This study evaluated CPE in aquatic environments and compared them to the local hospital isolates in Sweden. Phenotypic and genotypic analyses of antibiotic resistance of environmental and clinical CPE were performed. The relatedness of the isolates and possible clonal dissemination was evaluated using phylogenetic and phyloproteomic analysis. Klebsiella oxytoca carrying carbapenemase genes (blaVIM-1, blaIMP-29) were isolated from wastewater and the recipient river, while K. oxytoca (blaVIM-1) and Klebsiella pneumoniae (blaVIM-1, blaOXA-48, blaNDM-1, blaKPC-3) were isolated from patients at the local clinics or hospital. The K. oxytoca classified as sequence type 172 (ST172) isolated from the river was genotypically related to two clinical isolates recovered from patients. The similarity between environmental and clinical isolates suggests the dispersion of blaVIM-1 producing K. oxytoca ST172 from hospital to aquatic environment and the likelihood of its presence in the community. This is the first report of CPE in aquatic environments in Sweden; therefore, surveillance of aquatic and hospital environments for CPE in other urban areas is important to determine the major transfer routes in order to formulate strategies to prevent the spread of MDR bacteria.

  • 3.
    Khan, Faisal Ahmad
    et al.
    Örebro University, School of Science and Technology. The Life Science Centre – Biology.
    Söderquist, Bo
    Örebro University, School of Medical Sciences.
    Jass, Jana
    Örebro University, School of Science and Technology. The Life Science Centre – Biology.
    Prevalence and Diversity of Antibiotic Resistance Genes in Swedish Aquatic Environments Impacted by Household and Hospital Wastewater2019In: Frontiers in Microbiology, ISSN 1664-302X, E-ISSN 1664-302X, Vol. 10, article id 688Article in journal (Refereed)
    Abstract [en]

    Antibiotic-resistant Enterobacteriaceae and non-lactose fermenting Gram-negative bacteria are a major cause of nosocomial infections. Antibiotic misuse has fueled the worldwide spread of resistant bacteria and the genes responsible for antibiotic resistance (ARGs). There is evidence that ARGs are ubiquitous in non-clinical environments, especially those affected by anthropogenic activity. However, the emergence and primary sources of ARGs in the environment of countries with strict regulations for antibiotics usage are not fully explored. The aim of the present study was to evaluate the repertoire of ARGs of culturable Gram-negative bacteria from directionally connected sites from the hospital to the wastewater treatment plant (WWTP), and downstream aquatic environments in central Sweden. The ARGs were detected from genomic DNA isolated from a population of selectively cultured coliform and Gram-negative bacteria using qPCR. The results show that hospital wastewater was a reservoir of several class B beta-lactamase genes such as bla(IMP)(-1), bla(IMP)(-2), and bla(OXA-23), however, most of these genes were not observed in downstream locations. Moreover, beta-lactamase genes such as bla(OXA-48), bla(CDX-M-8), and bla(SFC-1), bla(VIM-1), and bla(VIM-13) were detected in downstream river water but not in the WWTP. The results indicate that the WWTP and hospital wastewaters were reservoirs of most ARGs and contribute to the diversity of ARGs in associated natural environments. However, this study suggests that other factors may also have minor contributions to the prevalence and diversity of ARGs in natural environments.

  • 4.
    Kumar, Ranjeet
    et al.
    School of Science and Technology, The Life Science Center-Biology, Örebro University, Örebro, Sweden.
    Pradhan, Ajay
    School of Science and Technology, The Life Science Center-Biology, Örebro University, Örebro, Sweden.
    Khan, Faisal Ahmad
    Örebro University, School of Science and Technology.
    Lindström, Pia
    Boliden Mineral AB, Boliden, Sweden.
    Ragnvaldsson, Daniel
    Envix Nord AB, Umeå , Sweden.
    Ivarsson, Per
    ALS Scandinavia AB, Täby, Sweden.
    Olsson, Per-Erik
    Örebro University, School of Science and Technology.
    Jass, Jana
    Örebro University, School of Science and Technology.
    Comparative analysis of stress induced gene expression in caenorhabditis elegans following exposure to environmental and lab reconstituted complex metal mixture2015In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 10, no 7, article id e0132896Article in journal (Refereed)
    Abstract [en]

    Metals are essential for many physiological processes and are ubiquitously present in the environment. However, high metal concentrations can be harmful to organisms and lead to physiological stress and diseases. The accumulation of transition metals in the environment due to either natural processes or anthropogenic activities such as mining results in the contamination of water and soil environments. The present study used Caenorhabditis elegans to evaluate gene expression as an indicator of physiological response, following exposure to water collected from three different locations downstream of a Swedish mining site and a lab reconstituted metal mixture. Our results indicated that the reconstituted metal mixture exerted a direct stress response in C. elegans whereas the environmental waters elicited either a diminished or abrogated response. This suggests that it is not sufficient to use the biological effects observed from laboratory mixtures to extrapolate the effects observed in complex aquatic environments and apply this to risk assessment and intervention.

  • 5.
    Yewale, Priti Prabhakar
    et al.
    Microbial Diversity Research Centre, Dr. D. Y. Patil Biotechnology and Bioinformatics Institute, Pune, India.
    Lokhande, Kiran Bharat
    Bioinformatics Research Laboratory, Pune, India.
    Sridhar, Aishwarya
    Biotechnology and Bioinformatics Institute, Pune, India.
    Vaishnav, Monika
    Biotechnology and Bioinformatics Institute, Pune, India.
    Khan, Faisal Ahmad
    Örebro University, School of Science and Technology.
    Mandal, Abul
    Systems Biology Research Center, School of Biosciences, University of Skӧvde, Skӧvde, Sweden.
    Swamy, Kakumani Venkateswara
    Bioinformatics Research Laboratory, Dr. D. Y. Patil Biotechnology and Bioinformatics Institute, Pune, India.
    Jass, Jana
    Örebro University, School of Science and Technology.
    Nawani, Neelu
    Microbial Diversity Research Centre, Dr. D. Y. Patil Biotechnology and Bioinformatics Institute, Pune, India.
    Molecular profiling of multidrug-resistant river water isolates: insights into resistance mechanism and potential inhibitors2019In: Environmental science and pollution research international, ISSN 0944-1344, E-ISSN 1614-7499Article in journal (Refereed)
    Abstract [en]

    Polluted waters are an important reservoir for antibiotic resistance genes and multidrug-resistant bacteria. This report describes the microbial community, antibiotic resistance genes, and the genetic profile of extended spectrum β-lactamase strains isolated from rivers at, Pune, India. ESBL-producing bacteria isolated from diverse river water catchments running through Pune City were characterized for their antibiotic resistance. The microbial community and types of genes which confer antibiotic resistance were identified followed by the isolation of antibiotic-resistant bacteria on selective media and their genome analysis. Four representative isolates were sequenced using next generation sequencing for genomic analysis. They were identified as Pseudomonas aeruginosa, Escherichia coli, and two isolates were Enterobacter cloacae. The genes associated with the multidrug efflux pumps, such as tolC, macA, macB, adeL, and rosB, were detected in the isolates. As MacAB-TolC is an ABC type efflux pump responsible for conferring resistance in bacteria to several antibiotics, potential efflux pump inhibitors were identified by molecular docking. The homology model of their MacB protein with that from Escherichia coli K12 demonstrated structural changes in different motifs of MacB. Molecular docking of reported efflux pump inhibitors revealed the highest binding affinity of compound MC207-110 against MacB. It also details the potential efflux pump inhibitors that can serve as possible drug targets in drug development and discovery.

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