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Background: Work-related fatigue is one of occupational health and safety problems that can be a risk factor for work-related accidents. ILO has recorded that every year, two million workers died because of work-related fatigue. There are many factors contributing to work-related fatigue, among others, age, sex, work load, work environment, and Carbon Monoxide exposure. The aim of the research was to observe the effect of Carbon Monoxide exposure and the characteristics of workers to work-related fatigue in workers in Fish Smoking Center, Wonosari Village, Demak Regency. Method: The research used cross sectional design using 42 samples. The data were analyzed using Multinomial Logistic Regression conducted on SPSS. Result: The research shows that 61.9% respondents experience work-related fatigue. Bivariate analysis suggests that age and sex aren-t significantly related to work-related fatigue, while employment period, work load and Carbon Monoxide exposure are significantly related to work-related fatigue. The result of multivariate analysis shows that work load (p=0,009) and Carbon Monoxide exposure (p=0,034) are significantly related to work-related fatigue. Work load is the dominant variable for work-related fatigue. Workers having heavy work load are exposed to work-related fatigue 12 times higher compared to those with light work load. Conclusion: The management of fish smoking center is expected to provide appropriate break and require the workers to wear mask during working hours.