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Biofuels produced from waste raw materials are used in combustion engines to reduce greenhouse gas (GHG) emissions, which are key to solving energy security problems and are the basis for sustainable economic development. Transesterification and hydrotreatment processes of vegetable waste oils enable the production of alternative fuels for powering compression-ignition engines. Different fuel production methods affect its physicochemical properties, which has a significant impact on the process of creating the mixture and the subsequent combustion process. This causes not always favorable changes in emissions and operating parameters of the diesel engine. In order to determine the level of these changes, comparative studies were carried out on the drive unit of an agricultural tractor in the form of a four-cylinder diesel engine equipped with a Common Rail injection system fueled with 25% mixtures of biodiesel and HVO with diesel oil and diesel oil as a reference. The measurements included analysis of engine operating parameters and regulated exhaust gas components at low, medium and high engine loads at 1500 and 2000 rpm. Additionally, cylinder pressure measurements are performed to gain insight into differences in combustion characteristics of the tested fuel blends. The addition of biodiesel causes significant reductions in PM emissions and ambiguous changes in NOx emissions. Emissions of nitrogen oxides (NOx), particulate matter (PM) and carbon dioxide (CO2) in the HVO25-fuelled engine showed reductions compared to diesel fuel.