The aim of this study was to identify the heavy metals forms (exchangeable and bound to carbonate, Fe/Mn oxides, bound to organic matter and sulphide, and residual) associated with different fractions of excess sludge produced by a membrane bioreactor (MBR). Furthermore, the release of metals from the sludge to the liquid was investigated by applying acid treatment using 10% (v/v) H2SO4 (T=25 degrees C, solid-liquid ratio 1:5 w/v) for contact time ranging from 15min to 4h. Metal partitioning in sludge, as determined by the sequential chemical extraction showed that the dominant form of both Ni and Zn was bound to the exchangeable and carbonate fraction; the latter were very unstable and sensitive to environmental conditions. The dominant Cu fraction was bound to organic matter and sulphide, while Pb was found to be mainly in the residual fraction which is very stable. Metal speciation after acidification with H2SO4 indicates changes of metal content in sludge and an increase of the exchangeable and bound to carbonate fraction for all metals except Cu. Acidification resulted in removal of 82% for Ni, 78% for Zn, 47% for Cu and 45% for Pb.