Food Safety Knowledge (FSK) among Food Handlers (FHS) in the food services in both hotels and hospitals has a crucial role in food protection and combating against foodborne diseases. The aim is to check the food safety (FS) training program upon FSK in the Jordanian hotels’ and hospitals’ food services. A pre-test-post-test design was used. A convenience sample of 412 food handlers from randomly selected hotels and hospitals were recruited. Data were collected using 118-questions Food Safety Knowledge Questionnaire (FSKQ); 40- questions tool for FS Practices. A significant improvement in FSK mean scores in the post-test (85.48±17.07/118 points corresponded to 72.44%) compared with the pretest (78.50±13.69/118 points, corresponding to 66.66%). The overall mean FSK score was considered to be “moderate” in pre and post-tests. FS Practice level mean was 26.86±4.38/40 points (67.14%), regarded as “moderate” in the pre-test and continued to be “moderate” after the training despite a significant increase in the post-test scores (27.92±4.45/40 point, corresponded to 69.80%, P=0.000). FHS worked in organizations where HACCP principles are implemented, possessed higher FSK and better FS practices. Although continued to be “moderate”, FSK and FS Practices among FHS in the Jordanian hotels and hospitals have been significantly improved after the FS Training Program.