Long-term preservation of anammox bacteria |
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Authors: | Jr" target="_blank">Michael J RothrockJr Matias B Vanotti Ariel A Szögi Maria Cruz Garcia Gonzalez Takao Fujii |
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Institution: | (1) USDA-ARS New England Plant, Soil, & Water Laboratory, Orono, ME 04469, USA;(2) USDA-ARS Coastal Plains Soil, Water, and Plant Research Center, Florence, SC 29501, USA;(3) Agriculture Technological Institute of Castilla and Leon (ITACyL), 47071 Valladolid, Spain;(4) Department of Applied Life Sciences, Sojo University, 4-22-1 Ikeda, Kumamoto 860-0082, Japan |
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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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