Background : The importance of healthcare workers' guidance for women infected with human immunodeficiency virus (HIV) regarding infant feeding practices cannot be overemphasised. Objective : To determine the knowledge, opinions and practices of healthcare workers in maternity wards in a regional hospital in Bloemfontein, Free State Province, South Africa, regarding infant feeding in the context of HIV. Methods : For this descriptive cross-sectional study, all the healthcare workers in the maternity wards of Pelonomi Regional Hospital who voluntarily gave their consent during the scheduled meetings (n ¼ 64), were enrolled and handed over the self-administered questionnaires. Results : Only 14% of the respondents considered themselves to be experts in HIV and infant feeding. Approximately 97% felt that breastfeeding was an excellent feeding choice provided proper guidelines were followed. However, 10% indicated that formula feeding is the safest feeding option. 45% stated that heat-treated breast milk is a good infant feeding option; however, 29% considered it a good infant feeding option but it requires too much work. Only 6% could comprehensively explain the term “exclusive breastfeeding” as per World Health Organisation (WHO) definition. Confusion existed regarding the period for which an infant could be breastfed according to the newest WHO guidelines, with only 26% providing the correct answer. Twenty per cent reported that no risk exists for HIV transmission via breastfeeding if all the necessary guidelines are followed. Conclusion : Healthcare workers' knowledge did not conform favourably with the current WHO guidelines. These healthcare workers were actively involved in the care of patients in the maternity wards where HIV-infected mothers regularly seek counselling on infant feeding matters.