This study assessed the training needs and knowledge gaps across five competency domains among the food protection staff of the Cincinnati Health Department. The five overarching competency domains assessed included 1) scientific knowledge; 2) foodborne illness knowledge, rules, and regulations; 3) temperature and storage; 4) inspection equipment; and 5) communication. A full network workforce assessment was conducted in a 3-year prospective longitudinal study design. Key findings show that competency areas identified as needing attention improved over time. The domain that consistently showed the highest percentage of workforce needing improvement was foodborne illness knowledge, rules, and regulations.