-In this study, physiological status of locomotive engineers were measured through EEG, ECG, EDA, PPG and respiration signals from 6 subjects to evaluate their arousal status during train operating. Existence of tunnels and mechanical vibration of train using 3-axes acceleration sensors were recorded simultaneously and were correlated with operator's physiological status. As the result of the analyzed subjects' physiological signals, mean SCR was increased in the section where more body movement is required. The RR interval was decreased before and after train stop due to the higher level of mental tension. The intensity of beta wave of EEG was found to be higher before and after train stop and tunnel section due to the increased mental arousal and tension. Therefore, it is expected that the outcomes of the physiological signals explored in this study can be utilized as the quantitative assessment methods for the arousal status to be used for sleepiness prevention system for vehicles operators which can greatly contribute to public transportation system safety.