© 2019. The Authors. Background: There is a high prevalence of human immunodeficiency virus (HIV), tuberculosis (TB), cancer and malaria in South Africa, and the drugs used to treat these conditions can be ototoxic. It is therefore important that healthcare professionals are able to identify and understand these drugs and their effects to ensure effective care of the patient. Objective: This study aimed to assess the knowledge regarding pharmacotherapy-induced ototoxicity amongst final year, medicine, pharmacy, audiology and nursing students across South African universities. Methods: A descriptive cross-sectional study design was used, and data were collected via a self-administered online questionnaire. Non-probability purposive sampling was used to identify the participants at the universities which train audiologists, pharmacists, medical and nursing students. Results: An overall response rate of 41% (n = 720) was obtained. Sixty-four per cent (n = 461) of respondents were women (median age: 23 years). The majority of the respondents (95%) knew what pharmacotherapy-induced ototoxicity was, but a few (39%) knew the general signs and symptoms of ototoxicity. Furthermore, just less than half of the sample (48%) could identify the specific ototoxic medicines and the type of damage caused by this medication. Conclusion: To manage pharmacotherapy-induced ototoxicity effectively, a multidisciplinary healthcare team must have sufficient knowledge about ototoxicity. Therefore, efforts should be made to introduce extensively concepts of pharmacotherapy-induced ototoxicity into the undergraduate curricula of pharmacy, medical, nursing and audiology programmes.