Microbial Ecology

, Volume 70, Issue 1, pp 118–131 | Cite as

Microbial Community Composition and Ultrastructure of Granules from a Full-Scale Anammox Reactor

  • Graciela Gonzalez-GilEmail author
  • Rachid Sougrat
  • Ali R. Behzad
  • Piet N.L. Lens
  • Pascal E. SaikalyEmail author
Environmental Microbiology


Granules in anammox reactors contain besides anammox bacteria other microbial communities whose identity and relationship with the anammox bacteria are not well understood. High calcium concentrations are often supplied to anammox reactors to obtain sufficient bacterial aggregation and biomass retention. The aim of this study was to provide the first characterization of bacterial and archaeal communities in anammox granules from a full-scale anammox reactor and to explore on the possible role of calcium in such aggregates. High magnification imaging using backscattered electrons revealed that anammox bacteria may be embedded in calcium phosphate precipitates. Pyrosequencing of 16S rRNA gene fragments showed, besides anammox bacteria (Brocadiacea, 32 %), substantial numbers of heterotrophic bacteria Ignavibacteriacea (18 %) and Anaerolinea (7 %) along with heterotrophic denitrifiers Rhodocyclacea (9 %), Comamonadacea (3 %), and Shewanellacea (3 %) in the granules. It is hypothesized that these bacteria may form a network in which heterotrophic denitrifiers cooperate to achieve a well-functioning denitrification system as they can utilize the nitrate intrinsically produced by the anammox reaction. This network may provide a niche for the proliferation of archaea. Hydrogenotrophic methananogens, which scavenge the key fermentation product H2, were the most abundant archaea detected. Cells resembling the polygon-shaped denitrifying methanotroph Candidatus Methylomirabilis oxyfera were observed by electron microscopy. It is hypothesized that the anammox process in a full-scale reactor triggers various reactions overall leading to efficient denitrification and a sink of carbon as biomass in anammox granules.


Anammox Autotrophic denitrification Heterotrophic denitrification Polygon-shaped bacteria Granules Microbial aggregates Granular sludge Pyrosequencing 



This work was supported by a Global Research Partnership–Collaborative Fellows Award (GRP-CF-2011-13-P) and discretionary investigator funds (P.E.S.) from King Abdullah University of Science and Technology. Special thanks are extended to Mohammed Alarawi and Siva Kumar Neelamegam at the Biosciences Core Laboratory at KAUST for generation of pyrosequencing and metagenomics reads, respectively, and to Krishna Katuri for providing M. barkeri and G. sulfurreducens cultures and Samik Bagchi for providing the nitrogen removal rate of the anammox granules.

Supplementary material

248_2014_546_MOESM1_ESM.docx (11.3 mb)
ESM 1 (DOCX 11553 kb)


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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Water Desalination and Reuse Center, Division of Biological and Environmental Sciences and EngineeringKing Abdullah University of Science and TechnologyThuwalSaudi Arabia
  2. 2.UNESCO–IHEDelftThe Netherlands
  3. 3.Advanced Nanofabrication Imaging and CharacterizationKing Abdullah University of Science and TechnologyThuwalSaudi Arabia

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