Background: Shift work (SW) can contribute to metabolic disorders affecting biological rhythms, inducing sleep deprivation, decreasing the resting metabolic rate, and changing dietary habits and eating behaviors. This study aims to assess the relationship of SW on physical and metabolic parameters, including the metabolic syndrome (MetS), and the contributing role of sleep quality, chronotype, chrononutrition, and the Mediterranean diet. Methods: Shift and nonshift workers underwent medical examinations and lab tests. MetS was diagnosed using the National Cholesterol Education Program’s Adult Treatment Panel III criteria. The Pittsburgh Sleep Quality Index, Chrononutrition Profile, Morning-Eveningness, and Prevention through Mediterranean Diet questionnaires were used to explore factors potentially affecting the development of metabolic disorders. Results: Shift (n = 172) and nonshift workers (n = 177), primarily employed in healthcare and social activities and in the education and administrative sectors, were enrolled. Mean systolic blood pressure levels were significantly higher in shift workers compared to the daytime counterpart (121.0 ± 12.9 vs.117.5 ± 13.2 mmHg, p = 0.013). MetS was significantly more frequent in the former group (25.6% vs.13.7%, p = 0.006). Higher percentages of shift workers compared to daytime workers, had poor quality of sleep (37.8% vs. 27.1%, p = 0.033), an evening chronotype (14% vs. 6.8%, p = 0.010), and high adherence to the Mediterranean diet (65.7% vs. 43.5%, p < 0.001), while no differences emerged for the chrononutrition profile. SW represents a risk factor for MetS, although its development results from the interaction of personal lifestyle, dietary habits, chronotype, chrononutrition, sleep quality, and occupational factors. Conclusion In the perspective of the Total Worker Health approach to workplace safety, health, and well-being, such an interplay should be addressed.