The viviparous gyrodactylid genus Macrogyrodactylus Malmberg, 1957 is endemic to Africa, composed of nine species from hosts of four freshwater fish families, including catfishes (Siluriformes: Clariidae). Three species, Macrogyrodactylus clarii Gussev, 1961; M. congolensis (Prudhoe, 1957) and M. karibae Douëllou et Chishawa, 1995, are primarily known to parasitise the African sharptooth catfish Clarias gariepinus (Burchell) in various African countries. From November 2017 to September 2019, a total of 184 individuals of C. gariepinus were collected from selected localities in southern Africa and their skin, fins and gills were surveyed for monogeneans. Three species of Macrogyrodactylus (M. clarii, M. congolensis and M. karibae) were found parasitising C. gariepinus from five localities in South Africa and Zambia. Overall prevalence was 50% to 100% with intensities of up to 109 parasites per host individual. New locality records in southern Africa, morphological observations and additional molecular data on the complete Internal Transcriber Spacer (ITS-1-5.8S-ITS-2) regions of the rDNA gene for the three gyrodactylid species are presented in this study.