Owing to high growth rate, being non-edible, and environmental friendliness; microalgae is a promising third generation biodiesel raw material. In this study, hydrogen and hydroxy gas aspirated compression ignition engine which was fuelled with microalgae biodiesel and low sulphur diesel fuel blend were investigated in order to evaluate their combined effect. The results showed that the brake power and torque output of the test engine decreased with microalgae biodiesel usage. Moreover, microalgae biodiesel addition results in lower carbon monoxide and nitrogen oxides emissions, and higher carbon dioxide. The introduction of hydrogen and hydroxy gas compensated the decrement of torque and power output and increment of carbon dioxide emission. The study enlightened that usage of microalgae biodiesel with hydrogen and hydroxy gas addition is a very promising combination from the environmental viewpoint. (C) 2018 Hydrogen Energy Publications LLC. Published by Elsevier Ltd. All rights reserved.