Traditional healers play an important role in southern Africa culture and health care including the HIV epidemic. Here we report among the first controlled studies of an HIV/AIDS, sexually transmitted infections (STI) and tuberculosis (TB) intervention for traditional healers in South Africa. At baseline 233 traditional healers were assessed in four selected communities in the KwaZulu-Natal province and received either an experimental intervention or a no intervention control condition. The intervention group received training in HIV/AIDS, STI, and TB prevention over 3.5 days as well as a supervisory follow-up visit. At 7-9 months follow-up intervention effects were significant for HIV knowledge and HIV and STI management strategies including conducting risk behavior assessments and counseling, condom distribution, community HIV/AIDS and STI education, and record keeping. The study found a high level of preparedness among traditional healers to work with and refer patients to biomedical health practitioners, yet no higher levels of referral to biomedical practitioners were found after the training.