Ammonia tolerant enriched methanogenic cultures as bioaugmentation inocula to alleviate ammonia inhibition in continuous anaerobic reactors

Ammonia is the most common inhibitor of anaerobic digestion (AD) process, resulting in suboptimal exploitation of the biogas potential of the feedstocks, causing significant economic losses to the biogas plants. Ammonia is mainly inhibiting the aceticlastic methanogens, while the hydrogenotrophic methanogens are more robust to ammonia toxicity effect. It has been shown that bioaugmentation of a pure strain of a hydrogenotrophic methanogen (i.e. Methanoculleus bourgensis) in an ammonia inhibited continuous anaerobic reactor can improve methane production more than 30%. Nevertheless, cultivation of a pure culture, to be used as bioaugmentation inoculum, poses technical difficulties due to the required sterile conditions and the special growing media. On the contrary acclimatized enrichment methanogenic cultures have lower requirements to sterility. In the present study, we used an enriched ammonia tolerant methanogenic culture as potential bioaugmentation inoculum in a continuous stirred tank reactor (CSTR) operating under “inhibited steady-state”, triggered by high ammonia levels (5 g NH4+-N L-1). The results of the current study established for the first time that bioaugmentation of an enriched ammonia tolerant methanogen in a CSTR reactor could completely alleviate the ammonia inhibitory effect. Furthermore, it was found that bioaugmentation with the enriched culture resulted in 25% higher methane production compared to when the bioaugmentation was achieved with pure methanogenic strains. The bioaugmentation was performed without pausing the continuous operation of the CSTR reactor and without excluding the ammonia-rich substrate from the feedstock. Thus, bioaugmentation with mixed methanogenic cultures could potentially support the development of an efficient and cost-effective biomethanation process of ammonia-rich organic waste in full-scale continuous reactors.