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The objective of this research project was to compare B20 (20% biodiesel fuel) and ultra-low-sulfur (ULSD) diesel-fueled buses in terms of fuel economy, vehicle maintenance, engine performance, component wear, and lube oil performance. We examined 15 model year (MY) 2002 Gillig 40-foot transit buses equipped with MY 2002 Cummins ISM engines. The engines met 2004 U.S. emission standards and employed exhaust gas recirculation (EGR). For 18 months, eight of these buses operated exclusively on B20 and seven operated exclusively on ULSD. The B20 and ULSD study groups operated from different depots of the St. Louis (Missouri) Metro, with bus routes matched for duty cycle parity. The B20- and ULSD-fueled buses exhibited comparable fuel economy, reliability (as measured by miles between road calls), and total maintenance costs. Engine and fuel system maintenance costs were also the same for the two groups after correcting for the higher average mileage of the B20 group.