Biofuels provide high oxygen content for combustion and do modify properties that influence the engine operation process such as viscosity, enthalpy of vaporization, and cetane number. Some requirements of performance, fuel consumption, efficiency, and exhaust emission are necessary for the validation of these biofuels for application in engines. This work studies the effects of the use of diethyl ether (DEE) in biodiesel-ethanol blends in a DI mechanical diesel engine. The blends used in the tests were B80E20 (biodiesel 80%-ethanol 20%) and B76E19DEE5 (biodiesel 76%-ethanol 19%-DEE 5%). Fossil diesel (D100) and biodiesel (B100) were evaluated as reference fuels. The results revealed similar engine efficiencies among tested fuels at all loads. The use of B100 increased CO and NOx and decreased THC compared to D100 at the three loads tested. B80E20 fuel showed an increase in NOx emission in comparison with all fuels tested, which was attributed to higher oxygen content and lower cetane number. THC and CO were also increased for B80E20 compared to B100 and D100. The use of B76E19DEE5 fuel revealed reductions in NOx and CO emissions, while THC emissions increased. The engine efficiency of B76E19DEE5 was also highlighted at intermediate and more elevated engine load conditions.