Applied Microbiology and Biotechnology

, Volume 100, Issue 12, pp 5339–5352 | Cite as

Microwave and ultrasound pre-treatments influence microbial community structure and digester performance in anaerobic digestion of waste activated sludge

  • Maria Westerholm
  • Sam Crauwels
  • Maarten Van Geel
  • Raf Dewil
  • Bart Lievens
  • Lise AppelsEmail author
Biotechnological products and process engineering


Comparative analyses of bacterial and archaeal community structures and dynamics in three biogas digesters during start-up and subsequent operation using microwaved, ultrasonicated or untreated waste activated sludge were performed based on 454 pyrosequencing datasets of part of 16S ribosomal RNA sequences and quantitative PCR. The pre-treatment increased the solubility, and thus the availability of the substrate for microbial degradation and significantly affected the succession of the anaerobic community structure over the course of the digestion. Bacteroidetes, Proteobacteria and Firmicutes were the dominant phyla in all digesters throughout operation. Proteobacteria decreased in relative abundance from 23–26 % to 11–13 % in association with enhanced substrate availability. Negative correlations between relative abundance of Alpha-, Beta- and Gammaproteobacteria and the substrate availability and/or biogas production were disclosed in statistical analyses. Clostridiales was the dominant order in Firmicutes, and Clostridiales, Clostridia and Firmicutes relative abundance and richness were shown to positively correlate with substrate availability and biogas generation. Methanogenic communities had a fairly restricted structure, highly dominated by Methanosaeta and Methanobrevibacter phylotypes. A gradual decline in Methanobrevibacter and increased representation of Methanosaeta concilii over time were particularly apparent in the digester receiving untreated waste activated sludge, whereas more diversified archaeal communities were maintained in the pre-treatment digesters. The quantitative PCR analyses revealed a methanogenic community distribution that coincided with the 454 pyrosequencing data.


Biogas Wastewater sludge Microbial dynamics and diversity Methanogenesis Next generation sequencing qPCR 



The authors would like to thank the Research Council of KU Leuven (projects OT/13/063 and F+/14/037) and the Industrial Research Council of KU Leuven (KP/10/006) for the financial support. We especially thank Sofie Houtmeyers for providing the digester samples, Stefan Ruyters for helping with the preparation of the 454 pyrosequencing run and Ken Meerbergen for assisting in some of the qPCR analyses.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2016_7321_MOESM1_ESM.pdf (835 kb)
Esm 1 (PDF 835 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Maria Westerholm
    • 1
  • Sam Crauwels
    • 2
  • Maarten Van Geel
    • 3
  • Raf Dewil
    • 1
  • Bart Lievens
    • 2
  • Lise Appels
    • 1
    Email author
  1. 1.Department of Chemical Engineering, Process and Environmental Technology LabKU LeuvenSint-Katelijne-WaverBelgium
  2. 2.Department of Microbial and Molecular Systems (M2S), Laboratory for Process Microbial Ecology and Bioinspirational Management (PME&BIM)KU LeuvenSint-Katelijne-WaverBelgium
  3. 3.Department of Biology, Plant Conservation and Population BiologyKU LeuvenHeverleeBelgium

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