Objective: We clarified the relationship between the degree of subjective fatigue, sleep, and physical activity among nurses working 16-hour night shifts in a rotating two-shift system. Materials and Methods: We investigated 15 nurses who were surveyed regarding their individual attributes, physical activity level (consumed calories), hours of sleep, sleep efficiency, sleep onset latency, sleep diary, and subjective symptoms. Nurses wore a Fitbit One (Fitbit Inc., San Francisco, CA, USA) for 7 consecutive days to measure sleep and physical activity. Results: Results were analyzed for nine participants, excluding those who withdrew or had missing data. The years of nursing experience, nurses' age, and the length of nocturnal awakening time of the high fatigue group were significantly longer than of the low fatigue group (p < .05). Years of nursing experience in the affiliated department of the high fatigue group was significantly shorter than of low fatigue group (p < .05). The number of nightshifts during the survey period was significantly higher in the high fatigue group than in the low fatigue group. Fatigue after work and body mass index (r = 0.46, p < .001), consumed calories (r = 0.30, p < .05), bedtime (r = -0.36, p < .05), and hours of sleep (r = -0.37, p < .01) were significantly correlated; however, the sleep indices were not correlated. Conclusion: We clarified that the degree of fatigue in nurses working 16-hour night shifts in a rotating two-shift system was related to individual factors, such as age, years of nursing experience, years of nursing experience in the affiliated department, number of nightshifts, and length of nocturnal awakening time. Nurses with greater fatigue had significant differences in their bedtime on days off and work days, which suggests that sleep rhythm may also affect fatigue.

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