This study is aimed at analyzing food safety knowledge and practices among food handlers in restaurants and street food markets in Dhaka, Bangladesh. Inadequate food handling practices remain a major worldwide health problem and are one of the main causes of food-related diseases. In Bangladesh, where the restaurant business is expanding quickly, food safety must be upheld to stop foodborne illness outbreaks. A cross-sectional study involving 300 street food vendors and restaurant workers was conducted through in-person interviews using a structured questionnaire. Descriptive and inferential statistical methods were employed to analyze the data. The study revealed that while most workers understood basic hygiene practices, critical knowledge gaps persisted. Male workers demonstrated lower food safety knowledge than females (p ≤ 0.01), and higher knowledge scores were associated with both greater education (p ≤ 0.001) and experience (p ≤ 0.05). Workers in fast food establishments showed higher knowledge levels (p ≤ 0.001), and knowledge was more strongly linked to job responsibilities than to training (p ≤ 0.001). In terms of practices, women adhered more closely to food safety guidelines than men (p ≤ 0.05). Age, education, and experience had no significant effects on practice adherence, though fast-food workers exhibited higher compliance (p ≤ 0.05). Training and job responsibilities showed no significant effects on food safety practices. The results demonstrate that there are disparities in food handlers’ understanding of food safety, with gaps primarily in the areas of cross-contamination avoidance, personal hygiene, and temperature control. Furthermore, observed practices suggest suboptimal adherence to advised food safety protocols.