Characterization of denitrifying phosphate-accumulating organisms cultivated under different electron acceptor conditions using polymerase chain reaction-denaturing gradient gel electrophoresis assay

Johwan Ahn, Tomotaka Daidou, Satoshi Tsuneda, Akira Hirata

    Research output: Contribution to journalArticle

    152 Citations (Scopus)

    Abstract

    To investigate the characteristics and the microbial diversity of denitrifying phosphate-accumulating organisms (DNPAOs) that are capable of conducting enhanced biological phosphorus removal (EBPR) using nitrate as electron acceptor, three sequencing batch reactors were operated under three different electron acceptor conditions, i.e., only oxygen, oxygen together with nitrate and only nitrate. Based on the chemical analysis concerning the biochemical transformation of each reactor, it was found that phosphate-accumulating organisms responsible for EBPR consisted of at least three populations including DNPAOs, and that the microbial community structure was changed according to the electron acceptor conditions. Also, the sludge cultivated with oxygen together with nitrate showed a drastic increase in the amount of phosphorus uptake under anoxic conditions, which suggested that a proportion of DNPAOs capable of utilizing nitrate under aerobic conditions were present. On the other hand, the change in microbial community structure depending on the type of electron acceptor was demonstrated by the analysis of the results of denaturing gradient gel electrophoresis of polymerase chain reaction-amplified 16S ribosomal DNA fragments. It was found that the bacteria commonly contained in all the reactors were Rhodocyclus sp. (96% identity) and Dechlorimonas sp. (97% identity) that belonged to the beta subclass of Proteobacteria on the basis of the analysis of the sequence excised from DGGE bands and the determination of phylogenetic affiliation. However, only the presence of Rhodocyclus sp. in all the reactors was demonstrated by fluorescent in situ hybridization analysis.

    Original languageEnglish
    Pages (from-to)403-412
    Number of pages10
    JournalWater Research
    Volume36
    Issue number2
    DOIs
    Publication statusPublished - 2002

    Fingerprint

    Polymerase chain reaction
    Electrophoresis
    polymerase chain reaction
    Nitrates
    Assays
    electrokinesis
    Phosphates
    Gels
    gel
    phosphate
    assay
    nitrate
    electron
    Electrons
    Phosphorus
    phosphorus
    oxygen
    Oxygen
    microbial community
    community structure

    Keywords

    • 16S ribosomal DNA
    • Denaturing gradient gel electrophoresis
    • Denitrifying phosphate-accumulating organisms
    • Enhanced biological phosphorus removal
    • Fluorescent in situ hybridization
    • Phosphate-accumulating organisms
    • Polymerase chain reaction

    ASJC Scopus subject areas

    • Earth-Surface Processes

    Cite this

    @article{a7dec0cb0fd245e99d6ee53a3647c22a,
    title = "Characterization of denitrifying phosphate-accumulating organisms cultivated under different electron acceptor conditions using polymerase chain reaction-denaturing gradient gel electrophoresis assay",
    abstract = "To investigate the characteristics and the microbial diversity of denitrifying phosphate-accumulating organisms (DNPAOs) that are capable of conducting enhanced biological phosphorus removal (EBPR) using nitrate as electron acceptor, three sequencing batch reactors were operated under three different electron acceptor conditions, i.e., only oxygen, oxygen together with nitrate and only nitrate. Based on the chemical analysis concerning the biochemical transformation of each reactor, it was found that phosphate-accumulating organisms responsible for EBPR consisted of at least three populations including DNPAOs, and that the microbial community structure was changed according to the electron acceptor conditions. Also, the sludge cultivated with oxygen together with nitrate showed a drastic increase in the amount of phosphorus uptake under anoxic conditions, which suggested that a proportion of DNPAOs capable of utilizing nitrate under aerobic conditions were present. On the other hand, the change in microbial community structure depending on the type of electron acceptor was demonstrated by the analysis of the results of denaturing gradient gel electrophoresis of polymerase chain reaction-amplified 16S ribosomal DNA fragments. It was found that the bacteria commonly contained in all the reactors were Rhodocyclus sp. (96{\%} identity) and Dechlorimonas sp. (97{\%} identity) that belonged to the beta subclass of Proteobacteria on the basis of the analysis of the sequence excised from DGGE bands and the determination of phylogenetic affiliation. However, only the presence of Rhodocyclus sp. in all the reactors was demonstrated by fluorescent in situ hybridization analysis.",
    keywords = "16S ribosomal DNA, Denaturing gradient gel electrophoresis, Denitrifying phosphate-accumulating organisms, Enhanced biological phosphorus removal, Fluorescent in situ hybridization, Phosphate-accumulating organisms, Polymerase chain reaction",
    author = "Johwan Ahn and Tomotaka Daidou and Satoshi Tsuneda and Akira Hirata",
    year = "2002",
    doi = "10.1016/S0043-1354(01)00222-6",
    language = "English",
    volume = "36",
    pages = "403--412",
    journal = "Water Research",
    issn = "0043-1354",
    publisher = "Elsevier Limited",
    number = "2",

    }

    TY - JOUR

    T1 - Characterization of denitrifying phosphate-accumulating organisms cultivated under different electron acceptor conditions using polymerase chain reaction-denaturing gradient gel electrophoresis assay

    AU - Ahn, Johwan

    AU - Daidou, Tomotaka

    AU - Tsuneda, Satoshi

    AU - Hirata, Akira

    PY - 2002

    Y1 - 2002

    N2 - To investigate the characteristics and the microbial diversity of denitrifying phosphate-accumulating organisms (DNPAOs) that are capable of conducting enhanced biological phosphorus removal (EBPR) using nitrate as electron acceptor, three sequencing batch reactors were operated under three different electron acceptor conditions, i.e., only oxygen, oxygen together with nitrate and only nitrate. Based on the chemical analysis concerning the biochemical transformation of each reactor, it was found that phosphate-accumulating organisms responsible for EBPR consisted of at least three populations including DNPAOs, and that the microbial community structure was changed according to the electron acceptor conditions. Also, the sludge cultivated with oxygen together with nitrate showed a drastic increase in the amount of phosphorus uptake under anoxic conditions, which suggested that a proportion of DNPAOs capable of utilizing nitrate under aerobic conditions were present. On the other hand, the change in microbial community structure depending on the type of electron acceptor was demonstrated by the analysis of the results of denaturing gradient gel electrophoresis of polymerase chain reaction-amplified 16S ribosomal DNA fragments. It was found that the bacteria commonly contained in all the reactors were Rhodocyclus sp. (96% identity) and Dechlorimonas sp. (97% identity) that belonged to the beta subclass of Proteobacteria on the basis of the analysis of the sequence excised from DGGE bands and the determination of phylogenetic affiliation. However, only the presence of Rhodocyclus sp. in all the reactors was demonstrated by fluorescent in situ hybridization analysis.

    AB - To investigate the characteristics and the microbial diversity of denitrifying phosphate-accumulating organisms (DNPAOs) that are capable of conducting enhanced biological phosphorus removal (EBPR) using nitrate as electron acceptor, three sequencing batch reactors were operated under three different electron acceptor conditions, i.e., only oxygen, oxygen together with nitrate and only nitrate. Based on the chemical analysis concerning the biochemical transformation of each reactor, it was found that phosphate-accumulating organisms responsible for EBPR consisted of at least three populations including DNPAOs, and that the microbial community structure was changed according to the electron acceptor conditions. Also, the sludge cultivated with oxygen together with nitrate showed a drastic increase in the amount of phosphorus uptake under anoxic conditions, which suggested that a proportion of DNPAOs capable of utilizing nitrate under aerobic conditions were present. On the other hand, the change in microbial community structure depending on the type of electron acceptor was demonstrated by the analysis of the results of denaturing gradient gel electrophoresis of polymerase chain reaction-amplified 16S ribosomal DNA fragments. It was found that the bacteria commonly contained in all the reactors were Rhodocyclus sp. (96% identity) and Dechlorimonas sp. (97% identity) that belonged to the beta subclass of Proteobacteria on the basis of the analysis of the sequence excised from DGGE bands and the determination of phylogenetic affiliation. However, only the presence of Rhodocyclus sp. in all the reactors was demonstrated by fluorescent in situ hybridization analysis.

    KW - 16S ribosomal DNA

    KW - Denaturing gradient gel electrophoresis

    KW - Denitrifying phosphate-accumulating organisms

    KW - Enhanced biological phosphorus removal

    KW - Fluorescent in situ hybridization

    KW - Phosphate-accumulating organisms

    KW - Polymerase chain reaction

    UR - http://www.scopus.com/inward/record.url?scp=0036132875&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=0036132875&partnerID=8YFLogxK

    U2 - 10.1016/S0043-1354(01)00222-6

    DO - 10.1016/S0043-1354(01)00222-6

    M3 - Article

    C2 - 11827346

    AN - SCOPUS:0036132875

    VL - 36

    SP - 403

    EP - 412

    JO - Water Research

    JF - Water Research

    SN - 0043-1354

    IS - 2

    ER -