Background: Breastfeeding is widely acknowledged as the optimal method of infant feeding. However, the percentage of South African mothers who exclusively breastfeed remains alarmingly low. Healthcare workers (HCWs) are pivotal in promoting breastfeeding; however, a deficiency in their knowledge, negative attitudes, and unfavourable practices can impede the promotion and support of breastfeeding. Aim: To assess the knowledge, attitudes, and support practices related to breastfeeding promotion of HCWs in the Motheo district, Free State province, compared with the 2016 WHO Guideline updates on HIV and infant feeding and 2018 Mother-Baby Friendly initiative guidelines. Methods: A quantitative analytical cross-sectional study involving paediatricians, obstetricians, general practitioners (GPs), and midwives was conducted using a self-administered questionnaire. Results: Over half of the HCWs (59%) were able to define exclusive breastfeeding (EBF), with a significant majority of GPs (76.1%) and midwives (78.6%) recommending EBF up to six months, compared with only 10% of paediatricians and 25% of obstetricians (p < 0.001). Some 11.1% of the total group of HCWs reported that breastfeeding is contraindicated for HIV-infected mothers, while 15.4% were unsure of the recommendation for breastfeeding among HIV-infected mothers. A higher proportion of midwives, compared with other HCWs, felt highly confident in providing breastfeeding advice (p < 0.001). Additionally, more than half (56%) of HCWs had not completed the 20-hour WHO Lactation Management Training, and 55% believed their prior breastfeeding training was not sufficient to support and educate breastfeeding mothers. Conclusion: Regular breastfeeding training of HCWs on the latest breastfeeding guidelines is necessary to equip HCWs with the knowledge and competence to successfully promote, protect, and support breastfeeding.