Spatial distribution and viability of nitrifying, denitrifying and ANAMMOX bacteria in biofilms of sponge media retrieved from a full-scale biological nutrient removal plant

The spatial distribution and activities of nitrifying and denitrifying bacteria in sponge media were investigated using diverse tools, because understanding of in situ microbial condition of sponge phase is critical for the successful design and operation of sponge media process. The bacterial consortia within the media was composed of diverse groups including a 14.5% Nitrosomonas spp.-like ammonia oxidizing bacteria (AOB), 12.5% Nitrobacter spp.-like nitrite oxidizing bacteria (NOB), 2.0% anaerobic ammonium-oxidizing (ANAMMOX) bacteria and 71.0% other bacteria. The biofilm appeared to be most dense in the relatively outer region of the media and gradually decreased with depth, but bacterial viabilities showed space-independent feature. The fluorescent in situ hybridization results revealed that AOB and NOB co-existed in similar quantities on the side fragments of the media, which was reasonably supported by the microelectrode measurements showing the concomitant oxidation of NH4 (+) and production of NO3 (-) in this zone. However, a significantly higher fraction of AOB was observed in the center than side fragment. As with the overall biofilm density profile, the denitrifying bacteria were also more abundant on the side than in the center fragments. ANAMMOX bacteria detected throughout the entire depth offer another advantage for the removal of nitrogen by simultaneously converting NH4 (+) and NO2 (-) to nitrogen gas.