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This paper presents results of research of Diesel engines emission of toxic substances in exhaust gases fuelled with a second generation biofuel BIOXDIESEL, which is a blend of Fatty Acid Ethyl Esters obtained from waste resources such waste vegetable and animal fats, bioethanol and standard Diesel fuel. Presented results are very promising, showing that the emission of toxic substances in exhaust gases are significantly reduced when fuelling with BIOXDIESEL fuel in comparison with standard Diesel fuel.