The purpose of this study was to observe heart rate and respiration responses to real traffic pattern flight. Nine experienced and nine less-experienced military pilots on active flying status participated in four uninterrupted traffic patterns flight missions with F-7 jet trainer. The heart rates and respiration waves were continuously recorded using a small recording device strapped around the chest. As compared with baseline values, significant increases in heart rates of the two groups (for experienced pilots group, F (11, 88) = 4.636, p = 0.000; for less-experienced, F (11, 88) = 4.437, p = 0.000) and mean respiration rates of less-experienced group (F (11, 88) = 4.488, p = 0.000) were obtained during the phases of take-off, final approach and landing. Heart rates of less-experienced pilots were significant higher than those of experienced pilots during the take-off phase (p < 0.05). There were no significant differences in respiration rates between the two groups at each phase of the whole flight. The results show that take-off, final approach and landing are the most mental workload phases in-flight, and less-experienced pilots show more mental workload than experienced pilots in take-off phase in-flight.