The World Health Organization (WHO) reports that food handlers play a significant role in the maintenance of food safety from food production to storage. This study aims to assess the implementation of WHO five keys to safer food in the food trucks in Jeddah city. This study was an observational study. A total of 103 registered and licensed food trucks in Jeddah were selected using the opportunity sampling method. The WHO’s five keys to safer food questionnaire was used by the researchers to inspect and evaluate the hygiene status of food trucks. A chi-square test was used to assess the association between the demographic characteristics of food truck handlers and the implementation of the variables used to evaluate WHO’s five keys to safer food in their food trucks. P value < 0.05 was considered significant. The participation rate was 64.4% (103/160). Most food truck handlers were older than 30 years (55.3%), were men (81.6%), had secondary-level education (40.8%), and were Saudi nationals (69.9%). More than 50% of food truck handlers had less than three years of experience in business, and 67% of food truck handlers had undergone training in food handling. The researchers found good implementation of Key 5c. – “Water from cans/bottles used for cooking” (93.2%), followed by Key 1a. “Cleaning and disinfecting done periodically” (78.6%), and poor implementation for Key 3a. – “Calibrated thermometers are used to check temperatures” (30.1%) in the food trucks. The overall implementation score of WHO’s five keys to safer food was found to be good (score ≥ 8) in 57.3% of food trucks. Food handlers’ age, gender, and training in food handling indicated a significant association with the implementation of WHO’s five keys to safer food. The exposure of food handlers to food handling training indicated a significant association with good implementation of all five keys to safer food.