In this study the optimization of the biogas yield from anaerobic co-digestion of manures and energy crops was carried out using four pilot scale CSTRs under different operating conditions. The effect on biogas yield of the partial substitution of energy crops with agro-waste was also investigated. For each substrate used during the continuous trials, BMP batch assays were also carried out to verify the maximum methane yield theoretically obtainable. Continuous operation results indicated that the co-digestion of manures, energy crops and agro-waste was viable at all operating conditions tested, with the greatest specific gas production of 0.54 m(3)/kg VSfed at an organic load rate of 2 kg TVS/m(r)(3)d consisting of 50% manure, 25% energy crops and 25% agro-waste on VS basis. No significant differences were observed between high and low loaded reactors suggesting the possibility of either improving the OLR in existing anaerobic reactors or reducing the design volumes of new reactors. (C) 2012 Elsevier Ltd. All rights reserved.