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The anaerobic digestion of three microalgae mixtures was evaluated at different substrate to inoculum (S/I) ratios (0.5, 1 and 3), biomass concentrations (3, 10 and 20gTS/kg) and pretreatments (thermal hydrolysis, ultrasound and biological treatment). An S/I ratio of 0.5 and 10gTS/kg resulted in the highest final methane productivities regardless of the microalgae tested (ranging from 188 to 395mL CH₄/gVS_added). The biological pretreatment supported negligible enhancements on CH₄ productivity, while the highest increase (46–62%) was achieved for the thermal hydrolysis. The optimum temperature of this pretreatment depended on the microalgae species. The ultrasound pretreatment brought about increases in CH₄ productivity ranging from 6% to 24% at 10,000kJ/kgTS, without further increases at higher energy inputs. The results here obtained confirmed the lack of correlation between the solubilization …