Introduction: Food handlers represent an occupational group which plays a major role in the spread of food borne diseases. The risk of food getting contaminated depends largely on the knowledge and practice of proper food hygiene measures among food handlers. Aim of work: To assess the knowledge and practice of food handlers towards food hygiene in addition to identifying the frequency of workers with positive nail culture. Materials and Methods: A cross sectional study including 140 subjects such that 54 worked in the kitchens of Ain Shams University Hospitals and 86 from 16 fast food restaurants were included in the study. Questionnaire and nail swabs for culture were done after obtaining their consent. Results: The study showed that the mean knowledge score was 76.6 ± 19.6 and the mean practice score was 68.1 ± 22.4. Knowledge questions related to the correct method of thawing food and the right place for vegetables and meat in the refrigerator showed less than 50% correct answers. Practice questions related to not keeping food at room temperature for more than 4 hours and not to continue working if having flu or hand lesions should total correct answers less than 50%. (40.7%) of those who agreed to give nail swabs showed positive culture with CoNS being the most frequently discovered pathogen. Conclusion: food handlers are an occupational group who need careful supervision and training courses on food hygiene and how to carry on their tasks safely without harming themselves or others.