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Microbial Ecology

Metabolic Profiles and Genetic Diversity of Denitrifying Communities in Activated Sludge after Addition of Methanol or Ethanol

Sara Hallin, Ingela Noredal Throbäck, Johan Dicksved, Mikael Pell
Sara Hallin
Department of Microbiology, Swedish University of Agricultural Sciences, Box 7025, SE 750 07 Uppsala, Sweden
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  • For correspondence: Sara.Hallin@mikrob.slu.se
Ingela Noredal Throbäck
Department of Microbiology, Swedish University of Agricultural Sciences, Box 7025, SE 750 07 Uppsala, Sweden
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Johan Dicksved
Department of Microbiology, Swedish University of Agricultural Sciences, Box 7025, SE 750 07 Uppsala, Sweden
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Mikael Pell
Department of Microbiology, Swedish University of Agricultural Sciences, Box 7025, SE 750 07 Uppsala, Sweden
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DOI: 10.1128/AEM.00809-06
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  • FIG. 1.
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    FIG. 1.

    Potential denitrification rates with 10 organic compounds supplied in excess (means ± standard deviations; n = 3) to generate metabolic profiles for samples from (a) the pilot-scale plants prior to ethanol (RP1) and methanol (RP2) addition and the full-scale plant without an external carbon source on two occasions (RF1 and RF2) and (b) the pilot-scale plants with ethanol (EP2) and methanol (MP2) addition on day 65 in comparison to the pilot-scale plants prior to the addition of ethanol (RP1) and methanol (RP2).

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    FIG. 2.

    Score plot (a) and loading plot (b) obtained by principal component analysis of potential denitrification rates with different electron donors to compare the metabolic profiles reported by Hallin and Pell (12) to those from this study, including those for the pilot-scale plants prior to ethanol (RP1) and methanol (RP2) addition, the full-scale plant without an external carbon source on two occasions (RF1 and RF2), and the pilot-scale plants with methanol (MP2) and ethanol (EP2) addition on day 65. Samples in score plot: P, pilot-scale plants; F, full-scale plants; E, addition of ethanol; M, addition of methanol; R, no external carbon source. Variables in loading plot: ME, methanol; ET, ethanol; AC, acetate; PR, propionate; BU, butyrate; GU, glucose; GY, glycerol; BE, benzoate; NA, no additional electron donor. Ellipses indicate processes with similar modes of operation.

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    FIG. 3.

    DGGE analysis with SYBR green staining of PCR-amplified partial nirK genes from the pilot-scale plant prior to methanol addition (RP2), the pilot-scale plant with methanol addition on day 21 (MP1) and day 65 (MP2), the pilot-scale plant with ethanol addition on day 28 (EP1) and day 65 (EP2), and the full-scale plant without external carbon addition at the start of the experiment (RF1) and on day 30 (RF2).

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    FIG. 4.

    Rarefaction curves showing the numbers of RFLP pattern types obtained from the numbers of nirK (white) and nirS (black) clones screened from the pilot-scale plant sludge prior to methanol addition (RP2; squares), after methanol adaptation on day 65 (MP2; circles), and after ethanol adaptation on day 65 (EP2; triangles) and the sludge from the full-scale plant without external carbon addition at the start of the experiment (RF1; diamonds). The solid line shows the maximum possible level of diversity.

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    FIG. 5.

    Distributions of RFLP pattern types represented by at least two nirK clones in (a) the pilot-scale plant prior to methanol addition (RP2), (b) the methanol-fed pilot-scale plant on day 65 (MP2), (c) the ethanol-fed plant on day 65 (EP2), and (d) the full-scale plant without an external carbon source at the start of the experiment (RF1). White pie slices represent RFLP pattern types specific for the sample, and the numbers refer to the RFLP pattern types.

  • FIG. 6.
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    FIG. 6.

    Distributions of RFLP pattern types represented by at least two nirS clones in (a) the pilot-scale plant prior to methanol addition (RP2), (b) the methanol-fed pilot-scale plant on day 65 (MP2), (c) the ethanol-fed plant on day 65 (EP2), and (d) the full-scale plant without an external carbon source at the start of the experiment (RF1). White pie slices represent RFLP pattern types specific for the sample, and the numbers refer to the RFLP pattern types.

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  • TABLE 1.

    Coverage of nirK and nirS clone libraries based on RFLP pattern typesa

    SampleLibrary coverage (%)
    nirKnirS
    RP28170
    MP28469
    EP28875
    RF19178
    • ↵ a RP2, sample from the pilot-scale plant prior to methanol addition; MP2, sample from the pilot-scale plant with methanol addition; EP2, sample from the pilot-scale plant with ethanol addition; RF1, sample from the full-scale plant without an external carbon source.

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    • Supplemental file 1 - Supplemental material (Phylograms of nirK and nirS genes)
      MS Word document, 79K.
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Metabolic Profiles and Genetic Diversity of Denitrifying Communities in Activated Sludge after Addition of Methanol or Ethanol
Sara Hallin, Ingela Noredal Throbäck, Johan Dicksved, Mikael Pell
Applied and Environmental Microbiology Aug 2006, 72 (8) 5445-5452; DOI: 10.1128/AEM.00809-06

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Metabolic Profiles and Genetic Diversity of Denitrifying Communities in Activated Sludge after Addition of Methanol or Ethanol
Sara Hallin, Ingela Noredal Throbäck, Johan Dicksved, Mikael Pell
Applied and Environmental Microbiology Aug 2006, 72 (8) 5445-5452; DOI: 10.1128/AEM.00809-06
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KEYWORDS

Ecosystem
Genetic Variation
Gram-negative bacteria
Nitrates
sewage

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