The CANON (Completely Autotrophic Nitrogen removal Over Nitrite) process was successfully developed in an air pulsing reactor type SBR fed with the supernatant from an anaerobic sludge digester and operated at moderately low temperatures (18-24 degrees C). The SBR was started up as a nitrifying reactor, lowering progressively the dissolved oxygen concentration until reaching partial nitrification. Afterwards, an inoculation with sludge containing Anammox biomass was carried out. Nitrogen volumetric removal rates of 0.25 g N L-1 d(-1) due to Anammox activity were measured 35 d after inoculation even though the inoculum constituted only 8% (w/w) of the biomass present in the reactor and it was poorly enriched in Anammox bacteria. The maximal nitrogen removal rate was of 0.45 g N L-1 d(-1). By working at a dissolved oxygen concentration of 0.5 mg L-1 in the bulk liquid, nitrogen removal percentages up to 85% were achieved. The reactor presented good biomass retention capacity allowing the accumulation of 4.5 g VSS L-1. The biomass was composed by ammonia oxidizing bacteria (AOB) forming fluffy structures and granules with an average diameter of 1.6 mm. These granules were composed by Anammox bacteria located in internal anoxic layers surrounded by an external aerobic layer where AOB were placed. (C) 2008 Elsevier B.V. All rights reserved.