Microbial Biofilms Formed in a Laboratory-Scale Anammox Bioreactor with Flexible Brush Carrier


This study is focused on investigation of biofilms formed in an anaerobic laboratory-scale bioreactor fed with medium for anammox bacteria oxidizing ammonia with nitrite. The mixed culture of anammox bacteria was enriched from the microbial community that sampled from the activated sludge of a denitrifying reactor at a wastewater treatment station located in the Sochi region, Russia. This community forms biofilms on the surface of the flexible polymer brush carriers, which are used for biomass immobilization in both laboratory and full-scale bioreactors. Anammox bacteria were discovered in the activated sludge community. The anammox community was enriched by incubation in an up-flow laboratory-scale anaerobic bioreactor with a flexible brush carrier. In the course of ~3 years, the loading rate of nitrogen substrates (ammonium and nitrite) increased from 100 to 5000 mg N L-1 day-1. The concentration of the substrates in the upper part of the reactor was 40 times less than in the influent. The pH values were 7.5 at the bottom and up to 9 in the upper part up of the reactor. Biofilms of two types developed in the reactor. Bunches of irregular spherical granules formed on the carrier filaments, while films of irregular thickness containing submerged spherical granules were formed on the walls of reactor. The anammox population was found to consist of at least three active species: a new strain of Candidatus Jettenia asiatica named “strain ecos” and two species of the genus Candidatus Brocadia. Other types of bacteria found in the community, including members of phylum Chloroflexi, were presumably involved in biofilm spatial organization.

Share and Cite:

Botchkova, E. , Litti, Y. , Kuznetsov, B. and Nozhevnikova, A. (2014) Microbial Biofilms Formed in a Laboratory-Scale Anammox Bioreactor with Flexible Brush Carrier. Journal of Biomaterials and Nanobiotechnology, 5, 76-82. doi: 10.4236/jbnb.2014.52010.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Strous, M., Fuerst, J.A., Kramer, E.H., Logemann, S., Muyzer, G., van de Pas-Schoonen, K.T., Webb, R., Kuenen, J.R. and Jetten, M.S. (1999) Missing Lithotroph Identified as New Planctomycete. Nature, 400, 446-449.
[2] Nozhevnikova, A.N., Simankova, M.V. and Litti, Yu.V. (2012) Application of the Microbial Process of Anaerobic Ammonium Oxidation (ANAMMOX) in Biotechnological Wastewater Treatment. Applied Biochemistry and Microbiology, 48, 667-684. http://dx.doi.org/10.1134/S0003683812080042
[3] Nozhevnikova, A.N., Litti, Yu.V., Nekrasova, V.K., Kulichevskaya, I.S., Grigoryeva, N.V., Kulikov, N.I. and Zubov, M.G. (2012) Anaerobic Ammonium Oxidation (Anammox) in Immobilized Activated Sludge Biofilms during the Treatment of Weak Wastewater. Microbiology, 81, 25-34.
[4] Litti, Yu.V., Nekrasova, V.K., Kulikov, N.I. and Nozhevnikova, A.N. (2013) Detection of Anaerobic Processes and Microorganisms in Immobilized Activated Sludge of a Wastewater Treatment Plant with Intense Aeration. Microbiology, 82, 690-697. http://dx.doi.org/10.1134/S0026261713060076
[5] Birnboim, H.C. and Doly, J. (1979) A Rapid Alkaline Extraction Procedure for Screening Recombinant Plasmid DNA. Nucleic Acids Research, 7, 1513-1523. http://dx.doi.org/10.1093/nar/7.6.1513
[6] Zhou, J., Bruns, M.A. and Tiede, J.M. (1996) DNA Recovery from Soils of Diverse Composition. Applied Environmental Microbiology, 62, 316-322.
[7] Krsek, M. and Wellington, E.M.H. (1999) Comparison of Different Methods for the Isolation and Purification of Total Community DNA from Soil. Journal of Microbiological Methods, 39, 1-16.
[8] Lane, D.J. (1991) 16S/23S Sequencing. In: Stackebrandt, E. and Goodfellow, M., Eds., Nucleic Acid Techniques in Bacterial Systematic, John Wiley & Sons, Ltd., Chichester, 115-175.
[9] Li, M., Hong, Y., Klotz, M.G. and Gu, J.-D. (2010) A Comparison of Primer Sets for Detecting 16S rRNA and Hydrazine Oxidoreductase Genes of Anaerobic Ammonium-Oxidizing Bacteria in Marine Sediments. Applied Microbiology and Biotechnology, 86, 781-790. http://dx.doi.org/10.1007/s00253-009-2361-5
[10] Sanger, F., Nicklen, S. and Coulson, A.R. (1977) DNA Sequencing with Chain-Terminating Inhibitors. Proceedings of the National Academy of Sciences, 84, 5463-5467. http://dx.doi.org/10.1073/pnas.74.12.5463
[11] Thompson, J.D., Higgins, D.G. and Gibson, T.J. (1994) CLUSTAL W: Improving the Sensitivity of Progressive Multiple Sequence Alignment through Sequence Weighting, Positions-Specific Gap Penalties and Weight Matrix Choice. Nucleic Acids Research, 22, 4673-4680. http://dx.doi.org/10.1093/nar/22.22.4673
[12] Strous, M., Kuenen, J.G. and Jetten, M.S.M. (1999) Key Physiology of Anaerobic Ammonium Oxidation. Applied and Environmental Microbiology, 65, 3248-3250.
[13] Cho, S., Takahashi, Y., Fujii, N., Yamada, Y., Saton, H. and Okabe, S. (2010) Nitrogen Removal Performance and Microbial Community Analysis of an Anaerobic Up-Flow Granular Bed Anammox Reactor. Chemosphere, 78, 11291135. http://dx.doi.org/10.1016/j.chemosphere.2009.12.034
[14] Kindaichi, T., Yuri, S., Ozaki, N. and Ohashi, A. (2012) Ecophysiological Role and Function of Uncultured Chloroflexi in an Anammox Reactor. Water Science Technology, 66, 2556-2561.

Copyright © 2022 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.