Training food handlers is essential to ensure food safety. However, the efficacy of training programs relying solely on theoretical information remains uncertain and often fails to induce significant changes in inadequate food practices. Training programs in good hygiene and food safety practices that integrate theoretical and practical approaches have emerged as a vital tool, enabling food handlers to apply their knowledge during work hours and clarify doubts. This study aimed to assess the impact of food safety training based on theoretical and on-the-job training on the microbiological counts of equipment, surfaces, utensils, and food handler (FH) hands. The hygiene and food safety conditions of four restaurants were analyzed through facility checklists, employee questionnaires, and microbiological analyses conducted before and after training. Eight sample collection moments were conducted at each restaurant before and after training. The pre-training results indicate that 15% and 26% of analyses for Enterobacteriaceae and total mesophilic aerobic bacteria (TMB), respectively, did not comply with hygiene safety limits. Additionally, 31% and 64% of Enterobacteriaceae and TMB values, respectively, exceeded safety limits on food handler hands. Positive cases of coagulase-positive Staphylococcus (CoPS) resulted from unprotected wounds on some FH hands. The presence of Listeria monocytogenes in drains was also identified as a concern. Following training, significant differences in results were observed. In many cases, there was a reduction of over 80% in microbial load for Enterobacteriaceae and TMB collected from equipment, surfaces, utensils, and food handler hands. The presence of L. monocytogenes in drains was also eliminated after food safety training. In conclusion, this study underscores the importance of effective training in improving food safety practices.