BACKGROUND: Efforts to combat Coronavirus disease (COVID-19) pandemic have significantly increased the quantity of bio-medical waste (BMW) generation. OBJECTIVES: A cross-sectional study was performed to assess the knowledge, practice, and attitude and factors affecting knowledge on BMW management among Health Care Workers (HCWs) when taking care of patients with COVID-19. METHODS: This cross-sectional study was conducted among 384 HCWs, actively involved in caring of COVID-19 patients in Healthcare settings of North 24 Parganas Health District, West Bengal. Data were collected using a structured self-administered questionnaire (through Google Forms) and an observational checklist after selection criteria and having consent. The Questionnaire included four sections like the sociodemographic information, knowledge related to BMW management, observational questionnaire assessing the practice of HCWs on BMW management and rating scale related to attitudes towards BMW management. RESULTS: 166 HCWs (43.2%) had an excellent knowledge with overall mean score 13.5 ± 3.6. A high mean score (14.4 ± 3.2) was obtained by doctors followed by nurses (13.6 ± 3.8). Regarding practice, the majority of HCWs (52.8%) followed the color-coding of BMW and 49.5% followed policies in separating the wastes according to hazard. Doctors (91%) and nurses (81%) had more favorable attitude than others. There was a statistically significant association found among knowledge level and educational qualification (P = 0.0001), gender (P = 0.001), and work experience (P = 0.05) and work area (P = 0.05). CONCLUSION: Emphasis should be given to aware and train all HCWs regarding proper BMW management during this pandemic to prevent infection transmission.