Mkambati Nature Reserve (NR) falls within the Pondoland Centre of Endemism, which is part of the Maputaland-Pondoland-Albany global biodiversity hotspot. The biodiversity status of this area is based largely on its flora, and the invertebrates are poorly known. The area is under threat from various proposed developments. We surveyed 14 orders in three invertebrate phyla at 26 sites with two main objectives: (1) to assess the fauna in terms of conservation value, and, (2) to identify habitats and sites of conservation concern. From the survey, 3231 samples were sent for identification and 425 species were identified. A minimum of 18 new species were confirmed. Mkambati NR shows exceptional diversity for molluscs (Gastropoda, 51 species), bees (Apoidea, 48 species) and true bugs (Heteroptera, 65 species). At least 43 species collected from the Reserve are South African endemics, 31 have a restricted distribution within South Africa and 18 are only known from the Reserve itself. Conservation implications: The authors provide the first assessment of the invertebrate fauna of the Mkambati NR, which indicates that it is a rich and important fauna. The results highlight the need to consider invertebrates in other biodiversity assessments in the Pondoland region. In terms of habitats, for both forest and grassland there was a large difference in the invertebrate communities at different sites, even over relatively short distances in grassland; shared habitat attributes clustered sites with more similar communities, for example, rocky ledges or the sea shore. All forest patches are a priority for protection.