Empowering a mesophilic inoculum for thermophilic nitrification: growth mode and temperature pattern as critical proliferation factors for archaeal ammonia oxidizers

Emilie N.P. Courtens, Tom Vandekerckhove, Delphine Prat, Ramiro Vilchez-Vargas, Marius Vital, Dietmar H. Pieper, Ken Meerbergen, Bart Lievens, Nico Boon, Siegfried E. Vlaeminck

Laboratory of Microbial Ecology and Technology (LabMET), Ghent University, Coupure Links 653, 9000 Gent, Belgium.

Abstract

Cost-efficient biological treatment of warm nitrogenous wastewaters requires the development of thermophilic nitrogen removal processes. Only one thermophilic nitrifying bioreactor was described so far, achieving 200 mg N L^-1 d^-1 after more than 300 days of enrichment from compost samples. From the practical point of view in which existing plants would be upgraded, however, a more time-efficient development strategy based on mesophilic nitrifying sludge is preferred. This study evaluated the adaptive capacities of mesophilic nitrifying sludge for two linear temperature increase patterns (non-oscillating vs. oscillating), two different slopes (0.25 vs. 0.08 °C d^-1) and two different reactor types (floc vs. biofilm growth). The oscillating temperature pattern (0.25 °C d^-1) and the moving bed biofilm reactor (0.08 °C d^-1) could not reach nitrification at temperatures higher than 46°C. However, nitrification rates up to 800 mg N L^-1 d^-1 and 150 mg N g^-1 volatile suspended solids d-1 were achieved at a temperature as high as 49°C by imposing the slowest linear temperature increase to floccular sludge. Microbial community analysis revealed that this successful transition was related with a shift in ammonium oxidizing archaea dominating ammonia oxidizing bacteria, while for nitrite oxidation Nitrospira spp. was constantly more abundant than Nitrobacter spp.. This observation was accompanied with an increase in observed sludge yield and a shift in maximal optimum temperature, determined with ex-situ temperature sensitivity measurements, predicting an upcoming reactor failure at higher temperature. Overall, this study achieved nitrification at 49°C within 150 days by gradual adaptation of mesophilic sludge, and showed that ex-situ temperature sensitivity screening can be used to monitor and steer the transition process.

Graphical abstract

Keywords: Archaea; thermophile; Nitrospira; Nitrososphaera.