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This paper describes the results of on-board measurement of engine performance and emissions in diesel vehicle operated with bio-diesel fuels. Here, two waste-cooking oils were investigated. One fuel is a waste-cooking oil methyl esters. This fuel is actually applied to a garbage collection vehicle with DI diesel engine (B100) and the city bus (B20; 80% gas oil is mixed into B100 in volume) as an alternative fuel of gas oil in Kyoto City. Another one is a fuel with ozone treatment by removing impurities from raw waste-cooking oils. Here, in order to improve the fuel properties, kerosene is mixed 70% volume in this fuel. This mixed fuel (i-BDF) is applied into several tracks and buses in Wakayama City. Then, these 3 bio-diesel fuels were applied to the on-board experiments and the results were compared with gas oil operation case.