Inland fisheries development in South Africa has been proposed to improve rural livelihoods and food security. However, inland fisheries are susceptible to collapse through overfishing. Therefore, establishing biological reference points for these fisheries is essential for long-term sustainability. Here, spawner-biomass-per-recruit (SBR) analysis was used to set biological reference points for a gillnet fishery targeting the rednose labeo Labeo rosae and Mozambique tilapia Oreochromis mossambicus at Flag Boshielo Dam on the Oliphants River in Limpopo Province, South Africa. Both species are currently exploited at the impoundment by subsistence fishers that keep their entire catch. The current yields are estimated at 59 and 172 g yr−1 for L. rosae and O. mossambicus, respectively. The per-recruit analysis showed that age-at-first-capture for both species should be 4 years, at minimum lengths of 280 mm TL and 250 mm TL for L. rosae and O. mossambicus, respectively, with fishers using a minimum mesh size of 100 mm to reduce the risk of stock collapse. To maintain a SBR value greater than 25% of a pristine population, the L. rosae yield could be increased to 98 g yr−1, while the O. mossambicus yield could be increased to 176 g yr−1, following implementation of the recommended mesh size and size limits. A per-recruit analysis for O. mossambicus that includes recruitment variability and error to accurately calculate the biological reference points is recommended.