Aims: 1) To assess and compare the knowledge and attitudes regarding biomedical waste (BMW) management in specialists, resident doctors, new medical interns, and final year nursing students. 2) To assess the effectiveness of a training program in changing the knowledge and attitudes regarding BMW management. Study Design: Stage 1-descriptive, Stage 2-quasi-experimental. Participants: Specialists, resident doctors, new medical interns, and final year nursing students. Setting: Tertiary hospital with attached medical college in Navi Mumbai. Data Collection tool: Pretested, precoded self-administered questionnaire. Intervention: Educational training program on BMW management, Period of Study: December 2010–March 2011. Statistical Analysis: Using software Statistical Package for Social Sciences (SPSS) version 20, chi-square, analysis of variance (ANOVA), Tukey’s post hoc, and Z tests applied. Results: There was a statistically signifi cant difference in the knowledge scores between the groups as determined by a one-way ANOVA test (F (3,226) = 11.098, P < 0.001). A Tukey’s post hoc test revealed that the specialists (20.82 ± 5.121) knowledge scores were signifi cantly higher as compared to resident doctors (16.96 ± 5.268), medical interns (18.44 ± 4.293), and nursing group (15.33 ± 5.144). The positive attitude towards safe management of BMW was not found to be signifi cant. After the training program in the medical interns’ a statistically signifi cant increase in their knowledge on BMW management was seen. Conclusion: The knowledge and attitudes between the groups of healthcare personnel varied and was not found to be satisfactory. Training programs with periodical sensitization sessions on BMW management are recommended, especially focusing at the junior level.