The sulphur-based bioleaching process using sulphur-oxidizing bacteria (SOB) has been demonstrated to be a feasible technology for removing heavy metals from contaminated sediments, but the excess sulphur application will lead to the re-acidification of bioleached sediments. The objective of the present study was to examine the effect of sulphur concentration on the bioleaching of heavy metals from contaminated sediments, with the ultimate purpose of minimizing the sulphur addition. The results showed that the inoculation of 7% of indigenous SOB, containing 3.6 x 10(8) colony forming units (CFU) mL(-1), and addition of elemental sulphur as a substrate (0.5 to 7.0 g L(-1)) resulted in a sharp decrease in sediment pH from an initial pH 8.0 to pH 1.4-2.4 and an increase in ORP (oxidation-reduction potential) from -10 mV to 500 mV within 10 days of bioleaching. Although the increase in sulphur concentration enhanced the rates of pH reduction and ORP elevation, the bioleaching process with the addition of 3.0 g L(-1) of sulphur was already sufficient to reach conditions of acidity (pH < 2.0) and ORP (500 mV) necessary for a satisfactory removal of metals, and, at day 10, 71.8% of Cu, 58.2% of Zn, and 25.3% of Cr were removed from the sediments. During the bioleaching process, Zn removal increased with a reduction in pH, whereas the removal of Cu and Cr increased not only with a reduction in pH but also with an increase in ORP. Results of sequential selective extraction indicated that the final levels of metal removals were dependent on their speciation distribution in the original sediments, and after bioleaching those unremoved metals in the bioleached sediments mainly existed in the residual fraction.