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Quantification of an Eikelboom type 021N bulking event with fluorescence in situ hybridization and real-time PCR

  • Environmental Biotechnology
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Abstract

Primers targeting 16S rRNA genes were designed to detect and quantify Eikelboom type 021N organisms by real-time PCR. Eikelboom type 021N filamentous bulking was induced in a laboratory-scale sequencing batch reactor and the evolution of Eikelboom type 021N 16S rRNA and 16S rRNA genes was monitored. A significant correlation was found between the sludge volume index and the amount of these filamentous organisms present in the sludge (r 2=94.6%, n=10, P<0.01), as measured by real-time PCR. The amount of Eikelboom type 021N 16S rRNA genes increased by a factor of 21 during the experiment, while the 16S rRNA increased by a factor of 33. Moreover, Eikelboom type 021N 16S rRNA increased with increased feeding frequency. It was observed that the RNA:DNA ratio peaked before the sludge volume index increased. In parallel, a fluorescence in situ hybridization study indicated a factor of four increase in the length of Eikelboom type 021N filaments, due to a factor of two increase in both length and number of Eikelboom type 021N filaments. Further, an increase in the fraction of filaments extending outside the activated sludge flocs was observed (19–55%). Monitoring of 16S rRNA genes and 16S rRNA of Eikelboom type 021N was shown to be valuable in evaluating activated sludge settling characteristics; and measuring RNA:DNA ratios may be used as an early warning tool for sludge bulking.

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Acknowledgements

This research was funded by a PhD grant of the Institute for the Promotion of Innovation through Science and Technology in Flanders (IWT–Vlaanderen). The authors thank Sylvie Seurinck, Roeland Grommen, and Tom Van De Wiele for critical comments.

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Authors and Affiliations

  1. Laboratory Microbial Ecology and Technology (LabMET), Ghent University, Ghent, Belgium

    Han Vervaeren, Jorg Matthys, Nico Boon & Willy Verstraete

  2. Aquafin NV Laboratorium, Spuimeerseweg 2, Hofstade, 9308, Belgium

    Kurt De Wilde

  3. Department of Civil and Environmental Engineering, University of Illinois at Urbana–Champaign, 3221 Newmark Civil Engineering Laboratory, 205 North Mathews Avenue, Urbana, IL 61801, USA

    Lutgarde Raskin

Authors
  1. Han Vervaeren
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  2. Kurt De Wilde
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  3. Jorg Matthys
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  4. Nico Boon
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  5. Lutgarde Raskin
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  6. Willy Verstraete
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Correspondence to Willy Verstraete.

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Vervaeren, H., De Wilde, K., Matthys, J. et al. Quantification of an Eikelboom type 021N bulking event with fluorescence in situ hybridization and real-time PCR. Appl Microbiol Biotechnol 68, 695–704 (2005). https://doi.org/10.1007/s00253-005-1963-9

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  • DOI: https://doi.org/10.1007/s00253-005-1963-9

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