Introduction: The practice of occupational health in the food industry is different from other industries as it protects both the health of the workers and of consumers. According to the WHO, in 2005, there were 1.8 million deaths from diarrhoea diseases, many from contamination of food and drinking water; 20% of which involved food handlers in food services. This study sought to assess and compare the knowledge, attitude and practices of, and the prevalence of intestinal parasitism between the food handlers in the University College Hospital, UCH, Ibadan and those in the government-owned secondary schools in Ibadan. Methodology: A comparative cross-sectional study was conducted among food handlers in the UCH and sampled secondary schools in Ibadan. A semi-structured, interviewer- administered questionnaire was used to obtain information on respondents’ socio-demographic characteristics, knowledge, attitude and practice of food hygiene. Stool samples were collected for microbiological examination for ova and parasites. Data were analysed using SPSS version 11. Result: A total of 270 food handlers were interviewed; 90 in the UCH and 180 in secondary schools. Their mean age was 34.49 + 12.69 (UCH) compared with 43.00 + 10.54 years (school food handlers). There were 24 (26.7%) males among the hospital food handlers compared to 2 (1.1%) among the school food handlers. Majority, 175 (97.2%) of the school food handlers were married compared to over half (54.4%) of the hospital food handlers (p = 0.000). Thirty one (34.4%) of the hospital food handlers compared with 7 (3.9%) of the school food handlers had higher education. Over half (51.1%) of the hospital food handlers compared with only 3 (1.7%) of the school food handlers had never had food handlers’ training (p = 0.000) while 73 (81.1%) of the hospital compared with none of the school food handlers had ever had food handlers’ test (Fisher’s exact = 0.000). Knowledge was good in both groups, 89 (98.9%) for hospital and 178 (98.9%) for school food handlers. The only area of poor knowledge was on disease pathogens involved in food poisoning where only 9 (10.0%) of hospital and 3 (1.7%) of school food handlers passed (Fisher’s exact = 0.003). Attitude was also good in both groups, 88 (97.8%) for hospital compared to 170 (94.4%) for school food handlers, except on attitude to not refreezing already defrosted food where 55 (61.1%) of hospital compared with 157 (87.2%) school food handlers had the wrong attitude (p = 0.000). Overall, food hygiene practice was good with all the respondents in both groups having self-reported good practice. Five (5.6%) hospital compared to 3 (1.7%) school food handlers had ascariasis (p = 0.076). There was no statistically significant association between intestinal parasitism and food handling practices. Effects of education and other factors could not be explored due to the small number of some variables. Conclusion and recommendation: This study showed that both hospital and school food handlers in Ibadan had good knowledge, attitude and practices of food hygiene; and both groups had low intestinal parasitism. The single most important factor affecting their knowledge, attitude and practices of food hygiene might have been educational status, though this could not be conclusively ascertained. Food handlers’ training should be continued and updated by the Oyo state government and it should be started by the UCH management.