Physically sedentary and overweight nurses appear to be less credible in inculcating healthy behaviour among patients. Shift-work has been shown to promote physical inactivity, whilst sedentarism strongly correlates with a high body mass index (BMI). We aim to determine the level of physical activity among hospital nurses of different work schedules, i.e. shift-work and day-work; and explore any associations between nurses’ demographic backgrounds, BMI, and work schedules with their physical activity level. This cross-sectional study design was employed. The self-administered Malaysian International Physical Activity Questionnaire – Long form (IPAQ-L) and a demographic survey sheet were provided to the eligible nurses. Bivariate and subsequent regression analyses were performed to determine their associations. A total of 1988 nurses from the University Malaya Medical Centre, Kuala Lumpur were invited to participate in this study. There were 1504 (76%) returned responses, with 77% of responding nurses working shift. Based on the IPAQ-L, 87% of the nurses were deemed highly active, while another 11% were moderately active. Upon stratifying into shift-work and day-work, statistically significant differences were observed between the groups in the domains of ‘work’ (p = 0.016), ‘domestic chores’ (p = 0.038), and in intensity-specific ‘walking’ (p = 0.046) and ‘vigorous’ activities (p = 0.034). There were no differences among groups within categories of physical activities (p = 0.355). Regression analysis showed significant difference for ‘duration of daily vehicle travel’, with the day-workers reporting a longer adjusted travel time (76.50 minutes/day, p < 0.001). Working shift does not seem to harm an individual nurse’s overall measured physical activity, as evidenced by equivalent high values of physical activity engagement between both work schedules. Any differences within domains and intensities of physical activities may be attributed to the respective cohort characteristics.