Introduction: The highest estimated cause of death in South East Asian Region (SEAR) attributes to diarrheal diseases (Global Food Infection Network, 2011). According to the CDC, 20 to 40% of foods borne illness associated with the consumption of contaminated food were from catering establishments. The purpose of the study was to assess the enteric pathogen profile and knowledge, attitude and behaviour regarding food hygiene among food handlers in our tertiary care center. Methods: Food handlers (n=123) were included in the study (June 2013- July2014) by non-random sampling after obtaining consent. Standardized WHO Five Keys for Safer Food questionnaire was used to assess the knowledge, attitude and behaviour on food hygiene among the food handlers. Stool specimens were collected for saline-iodine wet mount examination for microscopic examination of parasite ova and cysts, culture for bacterial enteropathogens and multiplex PCR for detection of Entamoeba histolytica. Results: The prevalence of stool pathogens in the hospital kitchen staff was 3.24%. The parasites detected were Entamoeba histolytica (1), Giardia intestinalis (2) and Blastocystis hominis (1) and the bacterium isolated was Campylobacter jejuni (1). Moderate level of knowledge (76.4%), favorable attitude (87%) and satisfactory self reported behaviour (92.7%) regarding food hygiene was observed. A significant positive correlation existed between attitude and self-reported behavior of food handlers (p<0.05). Conclusion: An overall prevalence of 5.12% of enteric pathogens was identified among the food handlers of JIPMER. Positive correlation between attitude and self-reported behaviour regarding food hygiene was observed among food handlers. Food hygiene behaviour was influenced by the work experience of the food handler.