Applied Microbiology and Biotechnology

, Volume 101, Issue 9, pp 3861–3869 | Cite as

Quantification of Chloroflexi Eikelboom morphotype 1851 for prediction and control of bulking events in municipal activated sludge plants in Japan

  • Tadashi NittamiEmail author
  • Lachlan B. M. Speirs
  • Takuya Yamada
  • Ichiro Suzuki
  • Junji Fukuda
  • Futoshi Kurisu
  • Robert J. Seviour
Environmental biotechnology


The dominant filamentous bacteria associated with bulking incidents in Japanese activated sludge plants with nutrient removal were identified and their quantitative correlations with sludge settleability were assessed, with the aim of controlling bulking incidents by specifically suppressing bacterial growth. Fluorescence in situ hybridization (FISH) analyses using existing oligonucleotide FISH probes indicated that the presence of Eikelboom type 1851 filamentous bacteria belonging to the phylum Chloroflexi is correlated with biomass settleability in the municipal wastewater treatment plants examined. Real-time quantitative PCR (qPCR) assays developed in this study also showed a linear correlation between type 1851 filament members and sludge settleability, with the exception of some winter samples. The real-time qPCR assays and 16S ribosomal RNA gene amplicon sequencing to reveal the microbial community of activated sludge showed that the abundance of type 1851 at 200 mL g−1 of sludge volume index was estimated to be about 1.9% of the total microbial cells. The abundance of type 1851 served as a bulking indicator in plants where type 1851 was dominant.


Amplicon sequence Fluorescence in situ hybridization (FISH) Type 1851/Kouleothrix Real-time qPCR Sludge volume index (SVI) 



The authors thank the Environmental Planning Bureau of the Yokohama City Government for providing the activated sludge and operational data for the wastewater treatment plants.

Compliance with ethical standards

This article does not contain any studies with human participants or animals performed by any of the authors. Informed consent was obtained from all individual participants included in the study.

Conflict of interest

The authors declare that they have no conflict of interest.


This research was supported by the Japan Society for the Promotion of Science (JSPS) KAKENHI Grant Numbers 23710087 and 15H02838. The authors also thank the Yokohama Academic Foundation for partial support of this study.


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Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  • Tadashi Nittami
    • 1
    Email author
  • Lachlan B. M. Speirs
    • 2
  • Takuya Yamada
    • 3
  • Ichiro Suzuki
    • 1
  • Junji Fukuda
    • 1
  • Futoshi Kurisu
    • 4
  • Robert J. Seviour
    • 2
  1. 1.Division of Materials Science and Chemical Engineering, Faculty of EngineeringYokohama National UniversityYokohamaJapan
  2. 2.Department of MicrobiologyLa Trobe UniversityBundooraAustralia
  3. 3.Department of Chemistry, Chemical Engineering and Life Science, College of Engineering ScienceYokohama National UniversityYokohamaJapan
  4. 4.Research Center for Water Environment TechnologyThe University of TokyoTokyoJapan

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