Objectives: The aim of this study was to compare nutrients intake, anthropometric, biochemical and psychiatric indices between shift working and day-time nurses. Methods: A cross-sectional comparative study was conducted in which ninety eight female nurses (55 day-time workers and 43 shift workers) from six educational hospitals of Jondi-Shapour University of medical sciences, Ahvaz, Iran were participated. A questionnaire including dietary, anthropometric, disease history and lifestyle pattern questions was completed and 3-day 24-hour recalls, Beck Depression Inventory (BDI) and Hospital Anxiety and Depression Scale (HADS) were collected. Serum hs-CRP and 25(OH)D 3 concentrations were measured by immunoturbidimetric and electrochemiluminescent immunoassay method, respectively. Anthropometric indices were measured according to World health organization standard protocol. Independent sample t and chi-square tests were used for statistical analysis. Results: There was a lower dietary intake of thiamin, riboflavin, niacin, folate, magnesium and iron in shift worker compared with day-time nurses (p < 0.05). No significant differences in serum hs-CRP concentrations, serum 25(OH)D 3 levels, vitamin D deficiency percentage, hemoglobin and hematocrite concentrations, and also anthropometric and psychiatric variables were found between two groups. Duration of exposure to sunlight was significantly higher in shift workers than in day time nurses. Engagement time in weekly physical exercise was around 11 times greater in day-time nurses compared with the shift work nurses (p = 0.001). Conclusions: This study showed that shift working is associated with some nutritional deficiencies and sedentary lifestyle among female nurses.