Microalgal biomass harvested from wastewater treatment high rate algal ponds may be valorised through anaerobic digestion producing biogas. However, microalgae anaerobic biodegradability is limited by their complex cell wall structure. Thus, pretreatment techniques are being investigated to improve microalgae methane yield. In the current study, thermal pretreatment at relatively low temperatures of 75-95 °C was effective at enhancing microalgae anaerobic biodegradability; increasing the methane yield by 70% in respect to nonpretreated biomass. Microscopic images showed how the pretreatment damaged microalgae cells, enhancing subsequent anaerobic digestion. Indeed, digestate images showed how after pretreatment only species with resistant cell walls, such as diatoms, continued to be present. Energy balances based on lab-scale reactors performance at 20 days HRT, shifted from neutral to positive (energy gain around 2.7 GJ/d) after thermal pretreatment. In contrast with electricity consuming pretreatment methods, such as microwave irradiation, thermal pretreatment of microalgae seems to be scalable. © 2014 American Chemical Society.