The European Union is promoting a new legislation through the "Proposal for a Directive of the European Parliament and of the Council on spreading of sludge on land", with special interest on human pathogens. Autothermal thermophilic aerobic digestion (ATAD) is described in this Proposal as an advanced treatment capable of producing sanitized sludge. A one-stage ATAD digester with effective volume of 15-m(3) was started up and studied over 19 months for its ability to stabilize and sanitize municipal sludge. Salmonella spp., Escherichia coli, and Clostridium perfringens spores were cultivated and pathogenity genes invA and cpa PCR-amplified. Volatile solids removal was 38.0% and the pathogen content significantly decreased by completely eliminating Salmonella spp. and E. coli but not Clostridium perfringens spores (9.6 x 10(3) spores mL(-1)). To completely achieve the disinfection of the sludge, a mesophilic stage was introduced after the ATAD treatment. Denaturing gradient gel electrophoresis analysis showed differences in the structures of the bacterial and fungal communities between thermophilic, mesophilic and raw sludge. The results demonstrated that the ATAD technology had the capability to produce sludge suitable for agricultural application when the operational parameters were stable and a mesophilic stage was introduced. (C) 2012 Elsevier B.V. All rights reserved.