Background: Biomedical waste is a neglected public health problem. Health care workers are the key for its management, so training is a must for an ideal performance. Aim: This study aimed to assess the current biomedical waste management process and the knowledge, attitude and practice of health care workers, develop and evaluate the effect of an improvement program. Subjects and Methods: pre-post intervention study was conducted at the fifteen PHC units affiliated to Ismailia health district and 307 healthcare workers for initial assessment and the development of the program. Then four PHC units and 80 HCWs participated for the implementation and evaluation of the program. Three Data collection tools were employed, the Individualized Rapid Assessment Tool (IRAT), a self-administered questionnaire to assess knowledge and attitude and an observation checklist for the practice. Results: The total average aggregate score percentage when rated using the I-RAT was 51% per urban and 47% for rural PHC unit. There are significantly higher correct knowledge mean percentage in young age than old age (p=0.013), according to their job and their departments (>0.001). Regarding the correct attitude there was a statistically significant difference between age, job, and department. As for practice, there are statistically significant differences between job and years of experience. The overall knowledge, attitude and practice mean percentage increased significantly in all the selected PHC units before and after the intervention with a p value (P<0.001). Conclusions: Training had a positive impact on staff knowledge, practices and attitudes.