The effect of oleate on the anaerobic digestion process was investigated. Two thermophilic continuously stirred tank reactors (CSTR) were fed with mixtures of cattle and pig manure with different total solid (TS) and volatile solid (VS) content. The reactors were subjected to increasing pulses of oleate. Following pulses of 0.5 and 1.0 g oleate/L, the most distinct increase in volatile fatty acid (VFA) concentrations were observed in the reactor with the lowest TS/VS content. This suggests a higher adsorption of oleate on the surfaces of biofibers in the reactor with the highest TS/VS and a less pronounced inhibition of the anaerobic digestion process. On the other hand, addition of 2.0 g oleate/L severely inhibited the process in both reactors, and a significant increase in all VFA concentrations combined with an immediate drop in methane production was noticed. However, 20 days after the reactors had been exposed to oleate both reactors showed a lower VFA concentration along with a higher methane production than before the pulses. This indicates that oleate had a stimulating effect on the overall process. The improved acetogenic and methanogenic activity in the reactors was confirmed in batch activity tests. In addition to this, toxicity tests revealed that the oleate pulses induced an increase in the tolerance level of acetotrophic methanogens towards oleate. When evaluating the usability of different process parameters (i.e., VFA and methane production) as indicators of process recovery, following the inhibition by oleate, propionate was found to be most suitable. (c) 2006 Wiley Periodicals, Inc.