Medical waste management (MWM) is of concern to the medical and general community. Adequate knowledge regarding management of healthcare waste is an important precursor to the synthesis of appropriate attitudes and practices of proper handling and disposal of medical waste by healthcare workers (HCWs).This study was designed to investigate knowledge, attitudes, and practices of doctors, nurses, laboratory technicians, and housekeeping staff, regarding MWM at a tertiary hospital in Gaborone, Botswana.This was a cross-sectional quantitative study using a self-administered questionnaire involving 703 participants. Data were analyzed using SAS software. Descriptive statistics were used to summarize the data. Responses for attitude of respondents were analyzed using nonparametric tests.The completion rate for this study was 90% with (632/703) questionnaires analyzed. Majority of respondents were nurses 60% (422/703), followed by housekeeping staff 24.3% (171/703), doctors 10.95% (77/703), and laboratory technicians 4.7% (33/703). The study showed that 66.9% (423/632) of respondents had some training in MWM, and 90.5% (572/632) claimed to have knowledge regarding the consequences of poor MWM, particularly health risks. There was a significant agreement among the respondents that segregation of medical waste should be done at the point of generation (mean score = 4.43 out of 5). Majority of respondents reported that the healthcare facility had a color-coding system (mean score = 4.59) and identified "lack of knowledge of the dangers of improper waste management by HCWs" as the major obstacle to MWM.This study showed that MWM practice at this facility was above average, although improvements were required in accessing waste disposal points and availability of personal protective equipment. Ongoing training should be provided to HCWs on MWM, with more attention to knowledge of regulatory requirements, and involvement of HCWs in development of MWM policies to enhance compliance.