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Long-term preservation of anammox bacteria

  • Applied Microbial and Cell Physiology
  • Published:
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Abstract

Deposit of useful microorganisms in culture collections requires long-term preservation and successful reactivation techniques. The goal of this study was to develop a simple preservation protocol for the long-term storage and reactivation of the anammox biomass. To achieve this, anammox biomass was frozen or lyophilized at two different freezing temperatures (−60°C and in liquid nitrogen (−200°C)) in skim milk media (with and without glycerol), and the reactivation of anammox activity was monitored after a 4-month storage period. Of the different preservation treatments tested, only anammox biomass preserved via freezing in liquid nitrogen followed by lyophilization in skim milk media without glycerol achieved stoichiometric ratios for the anammox reaction similar to the biomass in both the parent bioreactor and in the freshly harvested control treatment. A freezing temperature of −60°C alone, or in conjunction with lyophilization, resulted in the partial recovery of the anammox bacteria, with an equal mixture of anammox and nitrifying bacteria in the reactivated biomass. To our knowledge, this is the first report of the successful reactivation of anammox biomass preserved via sub-zero freezing and/or lyophilization. The simple preservation protocol developed from this study could be beneficial to accelerate the integration of anammox-based processes into current treatment systems through a highly efficient starting anammox biomass.

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Acknowledgements

The authors would like to thank Aprel Ellison, Kirsten Hiortdahl, and Laura Smith for their assistance in liquid sample and FISH analyses. This research was part of USDA-ARS National Program 206: Manure and By-Product Utilization: ARS Project 6657-13630-003-00D “Innovative Animal Manure Treatment Technologies for Enhanced Environmental Quality.”

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Authors and Affiliations

  1. USDA-ARS New England Plant, Soil, & Water Laboratory, Orono, ME, 04469, USA

    Michael J. Rothrock Jr

  2. USDA-ARS Coastal Plains Soil, Water, and Plant Research Center, Florence, SC, 29501, USA

    Matias B. Vanotti & Ariel A. Szögi

  3. Agriculture Technological Institute of Castilla and Leon (ITACyL), 47071, Valladolid, Spain

    Maria Cruz Garcia Gonzalez

  4. Department of Applied Life Sciences, Sojo University, 4-22-1 Ikeda, Kumamoto, 860-0082, Japan

    Takao Fujii

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  1. Michael J. Rothrock Jr
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  2. Matias B. Vanotti
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  3. Ariel A. Szögi
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  4. Maria Cruz Garcia Gonzalez
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  5. Takao Fujii
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Correspondence to Matias B. Vanotti.

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Rothrock, M.J., Vanotti, M.B., Szögi, A.A. et al. Long-term preservation of anammox bacteria. Appl Microbiol Biotechnol 92, 147–157 (2011). https://doi.org/10.1007/s00253-011-3316-1

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  • DOI: https://doi.org/10.1007/s00253-011-3316-1

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