Accumulation of substrate onto the biomass was quantified under steady-state and shock conditions in a fixed bed reactor fed with an oleic acid-based synthetic effluent. The accumulation of substrate onto the sludge was more dependent on oleic acid concentration than on oleic acid loading rate and decreased the acetogenic, acetoclastic and hydrogenophilic activity. However, even when the methanogenic activity measurements indicate a severe inhibition, the anaerobic sludge was able to methanise efficiently the accumulated substrate that was mainly adsorbed LCFA. Methanogenic activity measurements for a sludge loaded with 2,861 mg COD/gVSS as LCFA, revealed that only hydrogenophilic activity was detected, whereas the methanogenic activities with acetate, propionate and butyrate as substrates were null. However the methanogenic activity of the same sludge after allowing the depletion of the adsorbed LCFA were significantly enhanced in the presence of all substrates, except in propionate. A discussion about the relative importance of metabolic inhibition and transport limitations for the anaerobic degradation of LCFA is launched.