South African estuarine systems are becoming increasingly altered by anthropogenic and environmental factors, but the consequences of such changes for these systems are still not fully understood. The most common approach for evaluating the ecological status of aquatic systems is studying their associated foodwebs. Due to their high abundance and important ecological role, the larvae of estuarine round herring Gilchristella aestuaria (Gilchrist, 1913) are key candidates for examining the foodweb structure and function of southern African estuaries. The foodwebs and trophic interactions of G. aestuaria larvae in six estuaries in South Africa were compared using larvae sampled in November 2013 and analysed using delta C-13 and delta N-15 and Bayesian isotopic mixing models. The main prey type for G. aestuaria larvae in all estuaries was zooplankton. We found a high similarity among the Kariega, Gamtoos, Great Fish and Sundays estuaries in terms of consumers and potential sources for both d13C and delta N-15 signatures. Significant differences were found in delta C-13 values among marine-dominated estuaries, such as the Kromme Estuary, and the more freshwater-dominated systems. In addition, in the Kromme Estuary particulate organic matter was very important in the diet of G. aestuaria larvae. Our results suggest that both food availability and physical environmental parameters strongly affect the diet and condition of G. aestuaria and, consequently, the entire foodweb in the system.