Biodiesel is an alternative fuel consisting of the alkyl monoesters of fatty acids from vegetable oils or animal fats. Previous research has shown that biodiesel-fueled engines produce less carbon monoxide, unburned hydrocarbons, and particulate emissions compared to diesel fuel. One drawback of biodiesel is that it is more prone to oxidation than petroleum-based diesel fuel. In its advanced stages, this oxidation can cause the fuel to become acidic and to form insoluble gums and sediments that can plug fuel filters. The objective of this study was to evaluate the impact of oxidized biodiesel on engine performance and emissions. A John Deere 4276T turbocharged DI diesel engine was fueled with oxidized and unoxidized biodiesel and the performance and emissions were compared with No. 2 diesel fuel. The neat biodiesels, 20% blends, and the base fuel (No. 2 diesel) were tested at two different loads (100 and 20%) and three injection timings (3 degrees advanced, standard; 3 degrees retarded). The tests were performed at steady-state conditions at a single engine speed of 1400 rpm. The engine performance of the neat biodiesels and their blends was similar to that of No. 2 diesel fuel with the same thermal efficiency, but higher fuel consumption. Compared with unoxidized biodiesel, oxidized neat biodiesel produced 15 and 16% lower exhaust carbon monoxide and hydrocarbons, respectively. No statistically significant difference was found between the oxides of nitrogen and smoke emissions from oxidized and unoxidized biodiesel.