This study was aimed to examine the food safety knowledge of food handlers in hotels' kitchen, to determine existing knowledge gaps in food safety, and to examine relationship between food safety knowledge and some sample characteristics such as gender, age, education level, professional experience, and past attendance to food safety training course. A total of 378 food handlers working in hotel kitchens, located at six different cities in Turkey, participated in the cross-sectional study. The food safety knowledge score of participants was average with 53.70%. Knowledge scores related the different food safety aspects including personal hygiene (53.60%), food hygiene (53.91%), cross contamination (61.13%), health problems that would affect food safety (52.14%), symptoms of foodborne illnesses (52.00%), HACCP (51.00%) and food allergy (50.89%) were found average level. The most striking result of this study is that although the number of employees receiving food safety training is considerably high (82.3%), the food safety knowledge score was found less than expected. When viewed from this aspect, this work is remarkable about examining into content and adequacy of food safety training in Turkey.