Treatment of poultry industry effluents produces wastewater sludge with high levels of organic compounds and pathogenic microorganisms. In this research, the thermal pre-treatment of poultry slaughterhouse sludge (PSS) was evaluated for low temperatures in combination with different exposure times as a pre-hydrolysis strategy to improve the anaerobic digestion process. Organic compounds solubilization and inactivation of pathogenic microorganisms were evaluated after treatment at 70, 80 or 90°C for 30, 60 or 90 min. The results showed that 90°C and 90 min were the most efficient conditions for solubilization of the organic compounds (10%). In addition, the bacteria populations and the more resistant structures, such as helminth eggs (HE), were completely inactivated. Finally, the thermal pre-treatment applied to the sludge increased methane yield by 52% and reduced hydraulic retention time (HRT) by 52%.