Rail transportation in North America consumes over 4 billion gallons of diesel fuel [1]. This is raising energy security and supply chain resilience concerns. Adopting renewable or alternative fuels is a practical approach to reduce petroleum dependence and improve supply security. The objective of this paper is to investigate the combustion and emission characteristics of biodiesel and renewable diesel as drop-in fuels without engine modification. In this study, a single-cylinder, four-stroke locomotive engine was employed to investigate the combustion and emissions characteristics of four fuels: conventional diesel No. 2, plant-based biodiesel, animal-based biodiesel, and renewable diesel. The experimental campaign was carried out under both part-load and full-load operating conditions, with injection duration adjusted to achieve the targeted engine load and speed. Results indicate that both biodiesel fuels and renewable diesel deliver comparable peak in-cylinder pressure and brake thermal. efficiency relative to No. 2 diesel, demonstrating their possible use as drop-in fuels. Reductions in smoke emissions were observed for both biodiesels and renewable diesel fuels. However, plant-based and animal-based biodiesels both showed increases in NOx emissions under part-load conditions. At full load, elevated exhaust gas recirculation (EGR) ratios suppressed NOx formation across fuels, limiting assessment of biodiesel-specific NOx effects. Among the fuels tested, renewable diesel provided an additional advantage: reduced CO₂ emissions compared to both biodiesels. This study suggests that renewable diesel is a promising option for rail applications, combining operational performance comparable to petroleum diesel with reduced smoke and CO₂ emissions. Biodiesel, while effective at reducing smoke, may require further strategies to control NOx emissions.