Background: Rotating night shift nurses (RNS) are risky to unhealthy eating habits due to irregular work schedules, stressful environments, and unhealthy food choices. This may lead to obesity, a higher risk of medical conditions, and decreased productivity. Objectives: To identify the effect of shift work on nutritional habits and obesity among female nurses. Methods: A cross-sectional study was conducted at Assiut University Hospitals, including 108 nurses (54 RNS and 54 day shifts only), using an interview questionnaire including food frequency questionnaire on eating habits for recall of last month's consumption. Body mass index (BMI), and waist circumference were also measured. Results: The mean age was 30.15±4.96 years. Regarding eating habits of the nurses, day workers only had significantly more daily eating frequency and regular breakfast (p<0.05). Among all nurses, fresh fruits or vegetables and sweetened hot drinks were the most frequent snacks (60.2%, 59.3%, respectively). Fatty fast meal was the only food more frequent in RNS than day work (20.4 % versus 7.4%). Both groups had similar food groups, protein source variety, diet quality, and Mediterranean index. Moreover, 74% of nurses working included in the study were overweight and obese, and 88% had waist circumference >88 cm, which is significantly prevalent among day workers. Conclusions: Despite the absence of difference in dietary consumption between RNS and day workers, except in fatty meals, most studied nurses were at high risk of developing obesity-related diseases based on their high waist circumference. The findings underscore the need for improving workplace nutritional habits.