Anisakis simplex (AS) is a cause of allergic sensitization and potential occupational risk is suggested in fishermen and workers assigned to fish processing and sale. A cross-sectional study was conducted in order to assess possible health effects of occupational exposure to AS in workers recruited from western Sicily fisheries sector. Social, demographic, and occupation-related data were collected. Serum total immunoglobulin E (IgE) and specific IgE levels to AS (threshold >0.35 kU/L) were determined by an fluoroimmunoassay technique. Ninety-four subjects with potential occupational exposure (fishmongers, fishermen, fish industry employees) were recruited. Specific AS IgEs were detected in 20.2% of the study population. AS IgE seroprevalence was elevated 6.7-fold (p = .03) among fishermen/sailors compared with fish industry workers. The study suggested the importance to adopt specific prevention strategies against exposure to AS in the occupational setting.