OBJECTIVES: Monitoring of adverse drug reactions (ADRs) and establishment of a system of pharmacovigilance is the need of the day to ensure patient safety and prevent drug related morbidity and mortality. The present study was conducted to assess the knowledge, attitude and practices of healthcare professionals regarding pharmacovigilance (PV) in the Bahawalpur district of the Punjab province of Pakistan. The respondents were also asked about their knowledge regarding Isosorbide 5-mononitrate tragedy in the country (2011–2012). METHODS: This was a non-experimental, descriptive, questionnaire based cross-sectional survey that was carried out in the Bahawal Victoria Hospital and the Civil Hospital between December 2014 and March 2015. Using the convenient sampling technique, all the consented healthcare professionals were asked to self-complete a purpose designed structured questionnaire. A scoring scheme was used to assess the knowledge of the healthcare professionals regarding PV. Counts and proportions were used to present the data. The relationship between the characteristics of the respondents and the median knowledge score was analyzed by using the Mann-Whitney-U test and the Kruskal Wallis test. A p-value of less than .05 was considered significant.    RESULTS: A total of 107 health care professionals, including 88 medical doctors and 19 pharmacists, participated in the study. The median (25^th, 75^th) knowledge score of the respondents was 8 (7, 11) out of a maximum possible score of 17. Most of the respondents exhibited positive attitude towards implementation of PV system in the healthcare sector. Of all, 24 (22.4%) respondents did not know the actual cause of death among the patients taking Isosorbide 5-mononitrate. The ADRs reporting and monitoring was not practiced routinely owing to multiple factors. CONCLUSIONS: After about three years of Isosorbide 5-mononitrate tragedy, our respondents still exhibited poor knowledge, and there was lack of PV related practices. However, positive attitude towards implementation of PV system is good omen.