Diesel engines are one of the most important power generating units these days. Increasing greenhouse gas emissions level and the need for energy security has prompted increasing research into alternative fuels for diesel engines. Biodiesel is the most popular amongst the alternatives for diesel fuel as it is biodegradable, renewable and can be produced domestically from vegetable oils. In recent years, hydro-treated vegetable oil (HVO) has also gained popularity due to some of its advantages over biodiesel such as higher cetane number, lower deposit formation, storage stability etc. HVO is a renewable, paraffinic biobased alternative fuel for diesel engines similar to biodiesel. Unlike biodiesel, the production process for HVO involves hydrogen as catalyst instead of methanol which removes oxygen content from vegetable oil. A modified 6-cylinder heavy-duty diesel engine (modified for operation with single cylinder) was used for studying particle number emission characteristics for HVO fuel. The investigation was performed for varying fuel injection pressure at various engine operating loads (6, 8, 10, 12 and 14 bar IMEP). Five rail pressures were chosen from 800 to 2000 bar at a step of 300 bar. The results show that increase in rail pressure tends to increase nucleation mode particle number concentration (quantify the increase) while increase in engine load results in higher total particle number concentration. No significant differences were observed in soot and oxides of nitrogen (NOx) emission for HVO compared to mineral diesel. The fraction of emitted particles in the nucleation mode was observed to increase with increasing fuel injection pressure.