The aim of this study was to assess the hygiene practices of food handlers in municipal schools of Natal, Brazil, where 27 public schools were evaluated, using a checklist and microbiological analysis of hands. It was found that 74.1% of the handlers did not receive periodic training, 51.9% did not undergo annual health examinations and 100% did not practice proper hand hygiene, a situation that reflected significantly (p < 0.05) in hand contamination, in which fecal coliforms were detected on 55.6% of the hands analyzed. It was concluded that the schools studied did not have appropriate hygienic conditions, suggesting the need for interventions that ensure the quality of school food served to the children.