Ammonia is the major inhibitor of anaerobic digestion (AD) process in biogas plants. In the current study, the bioaugmentation of the ammonia tolerant SAO co-culture (i.e. Clostridium ultunense spp. nov. in association with Methanoculleus spp. strain MAB1) in a mesophilic up-flow anaerobic sludge blanket (UASB) reactor subjected to high ammonia loads was tested. The co-cultivation in fed-batch reactors of a fast-growing hydrogenotrophic methanogen (i.e. Methanoculleus bourgensis MS2(T)) with the SAO co-culture was also investigated. Results demonstrated that bioaugmentation of SAO co-culture in a UASB reactor was not possible most likely due to the slow maximum growth rate (mu(max) = 0.007 h(-1)) of the culture caused by the methanogenic partner. The addition of M. bourgensis to SAO led to 42% higher growth rate (mu(max) = 0.01 h(-1)) in fed-batch reactors. This indicates that methanogens were the slowest partners of the SAO co-culture and therefore were the limiting factor during bioaugmentation in the UASB reactor. (c) 2013 Elsevier Ltd. All rights reserved.