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Stable isotope analyses, specifically δ13C and δ15N, are useful tools increasingly used to understand ecosystem function, food web structures, and consumer diets. Although the iconic tigerfish Hydrocynus vittatus is regarded as an apex predator in southern African freshwater systems, little information is available regarding their feeding behavior and how this may change with growth or differ between ecosystems, with most information stemming from stomach content analyses (SCA). The aim of the present study was to address this lack of information through a baseline study of the diet of large and small tigerfish in various lentic and lotic ecosystems in South Africa using stable isotope methods. Fish and various food web components and food sources were collected from two river and two lake ecosystems in South Africa. The δ13C and δ15N values for all samples were determined and multivariate analyses and Bayesian analytical techniques applied to determine the feeding ecology of H. vittatus and how this may differ with size and habitat type. Analyses revealed a substantial difference in the type and abundance of food sources contributing to the diet of H. vittatus between ecosystems, most prominently between the lotic systems, where less dietary specialization was observed, and lentic systems where more specialization was observed. Furthermore, there was a distinct difference in diet between small and large tigerfish, especially in the lotic system, indicating an ontogenetic diet shift as tigerfish grow and further supporting previous SCA studies. This is the first study of its kind on the African continent for H. vittatus and the findings illustrate the value of stable isotope analysis in providing in‐depth information into the feeding ecology of consumers and how this may differ between size classes and habitat types.