The effect of a pulse addition of vitamin B12 as cobalt source to restore the performance of cobalt depleted methanol-fed bioreactors was investigated. One upflow anaerobic sludge bed (UASB) reactor was supplied with a pulse of vitamin B12, and its operation was compared to that of another cobalt depleted UASB reactor to which a pulse of CoCl2 was given. The addition of cobalt in the form of CoCl2 supplies enough cobalt to restore methanogenesis and maintain full methanol degradation coupled to methane production during more than 35 days after the CoCl2 pulse. Similar to CoCl2, pulse addition of vitamin B12 supplies enough cobalt to maintain full methanol degradation during more than 35 days after the pulse. However, the specific methanogenic activities (SMAs) of the sludge in the vitamin B12 supplied reactor were around 3 times higher than the SMA of the sludge from the CoCl2 supplied reactor at the same sampling times. An appropriate dosing strategy (repeated pulse dosing) combined with the choice of vitamin B12 as the cobalt species is suggested as a promising dosing strategy for methanol-fed anaerobic bioreactors limited by the micronutrient cobalt. © 2010 Elsevier Ltd. All rights reserved.