Objectives: The proper porthole angle contributes to relieving the operation fatigue and improving the efficiency of oceanauts. In this study, the authors explored the effect of 3 different porthole longitudinal positions on the oceanauts’ back muscles using surface electromyography (sEMG) analysis, and the characteristics of the perceived body comfort was obtained. Material and Methods: Overall, 40 healthy participants were recruited to perform tasks in a simulated cabin environment. Electromyographic (EMG) signals were recorded from the trapezius medius, lower trapezius, and erector spinal muscles for porthole angles of –5°, –15°, and +15°, relative to the horizontal line of sight during a 21-minute experiment. The subject comfort scores were collected at 7, 14 and 21 min. The integrated electromyogram (iEMG) and the root mean square (RMS) of EMG signals, as well as the mean power frequency (MPF), and the mean frequency (MF) were calculated. Results: The subjective scores of the +15° porthole at each stage of work are higher than those of the –15° and –5° portholes. The results of iEMG, RMS, MF and MPF all indicated that the +15° porthole design was more conducive to lowering the rate of muscle fatigue, while the –5° and –15° portholes increased the muscle fatigue rate and led to greater fatigue. It was found that the lower trapezius was more prone to fatigue than the trapezius medius and erector spinal muscles. The height, weight and body mass index of the participants were found to negatively correlate with muscles at the +15° porthole, which is highly consistent with the actual situation. Conclusions: The findings suggested that the +15° position was optimal for delaying the muscle fatigue of the participants and for improving the work efficiency of oceanauts. Int J Occup Med Environ Health. 2021;34(6):701–21