Although use of organochlorine pesticides (OCPs) has been banned or severely limited on a global basis, concerning concentrations continue to be reported in many tropical and subtropical regions of the world. These habitats often support high levels of unique biodiversity and vulnerable communities that depend on the environment for their survival. We investigated threats associated with OCP contamination at Lake St Lucia, a global hotspot for biodiversity and a UNESCO World Heritage Site in South Africa. Lake St Lucia is sustained largely by surface runoff from catchment areas where significant quantities of OCPs have historically been used in agriculture and where DDT continues to be used for disease control. Sediments (n = 40) collected from the two largest fluvial inputs to Lake St Lucia showed that these rivers represent important sources of contaminants, with ∑OCP concentrations ranging between 74 and 510 ng g-1. Measured HCH, dieldrin, ∑DDT and ∑chlor concentrations exceeded NOAA sediment toxicity guidelines in the majority of samples analysed. Bioaccumulation was assessed by examining residue concentrations in muscle tissues from two abundant fish species from Lake St Lucia. OCPs were detected in all samples analysed, with total concentrations ranging 860-5000 ng g-1 lw and 390-3200 ng g-1 lw for Oreochromis mossambicus (n = 17) and Clarias gariepinus (n = 41), respectively. A health risk assessment indicated potential dietary risk associated with exposure to aldrin, dieldrin and heptachlor, although the cumulative effect of OCPs on human health, ecosystem biodiversity and long-term ecotourism sustainability remains unknown.