Development of a super high-rate Anammox reactor and in situ analysis of biofilm structure and function

The anaerobic ammonium oxidation (Anammox) process is a new efficient and cost effective method of ammonium removal from wastewater. Under strictly anoxic condition, ammonium is directly oxidised with nitrite as electron acceptor to dinitrogen gas. However, it is extremely difficult to cultivate Anammox bacteria due to their low growth rate. This suggests that a rapid and efficient start-up of Anammox process is the key to practical applications. To screen appropriate seeding sludge with high Anammox potential, a real-time quantitative PCR assay with newly designed primers has been developed. Thereafter, the seeding sludge with high abundance of Anammox bacteria (1.7 × 10⁸ copies/mg-dry weight) was selected and inoculated into an upflow anaerobic biofilters (UABs). The UABs were operated for more than 1 year and the highest nitrogen removal rate of 24.0 kg-N m⁻³ day⁻¹ was attained. In addition, the ecophysiology of Anammox bacteria (spatial distribution and in situ activity) in biofilms was analysed by combining a full-cycle 16S rRNA approach and microelectrodes. The microelectrode measurement clearly revealed that a successive vertical zonation of the partial nitrification (NH₄⁺ to NO₂⁻), Anammox reaction and denitrification was developed in the biofilm in the UAB. This result agreed with the spatial distribution of corresponding bacterial populations in the biofilm. We linked the micro-scale information (i.e. single cell and/or biofilm levels) with the macro-scale information (i.e. the reactor level) to understand the details of Anammox reaction occurring in the UABs.