Acid mine drainage (AMD) is a serious environmental problem resulting from extensive sulphide mining activities. The old copper mine of S. Domingos in Southeast Portugal is an example of such a situation. The abandoned open-pit from the mining operations resulted in the creation of a large pit lake with acidic water (pH∼2) and high contents of sulphate and heavy metals. Sulphatereducing biological processes have been studied as a remediation technology for this problem. A new application based on a simple and semi-continuous process for the treatment of S. Domingos AMD has been presented herein. Experiments using bench scale fixed-bed column bioreactors were carried out to evaluate the efficiency of the process. Sewage, anaerobic sludge and soil from the mining area were tested as solid matrices and/or inocula, as well as sources of complex organic substrates. The addition of lactose as a supplementary carbon source, easily available at zero cost or at negative cost in the effluents of the local cheese industries, was also tested. The data obtained indicate that it is possible to use the matrices tested for the production of sulphide by sulphate reduction, and that the regular addition of lactose is effective. Results showed that the process is efficient for the precipitation of the main dissolved metals, for the reduction in the sulphate content and, most importantly, for the neutralization of the AMD. Moreover, the use of soil as solid support also showed the possibility of using this process for the decontamination of both waters and soils.