Aims: The therapeutic potential of honey for the treatment of wound infections is well documented. However, South African (SA) honey has been poorly explored as an antimicrobial agent and given the well-established antimicrobial properties of the indigenous plant species from SA, there is the potential that honey from this geographical region may exhibit noteworthy anti-infective properties. In this study, the antimicrobial properties of 42 SA honey samples were determined. Methods and Results: The minimum inhibitory concentration (MIC) agar dilution method was used to determine antimicrobial activity. The MICs of the honeys ranged from 6.25 to 50.00%. Samples 4-(CITYMIX/WC), 12-(BUSHVELD/KZN), 15-(ONION/WC), 16-(FYNBOS/WC), 17-(AKMS/FS), 19-(CITYMIX/FS), 41-(INDIGENOUS/WC) and 52-(SURBURBANGARDEN/WC) displayed broad-spectrum antimicrobial activity. The physicochemical properties including pH, water content and stable isotope analysis (SIA) was analysed. The pH of the honeys ranged between 3.89 and 5.09. The SIA revealed strong overall trends between protein concentration and MIC suggesting close links with antimicrobial activity. Conclusion: A number of SA honey samples tested have potential as an effective antimicrobial agent in wound healing. Significance and Impact of the Study: The future of South Africa's market for medical grade and therapeutic honeys looks promising as the antimicrobial properties of the honeys have some superior activity.