Successful aquaculture species are often chosen for their fast growth rates and fecundity, which are also characteristics of invasive species. The Pacific oyster Crassostrea gigas, which constitutes 80% of global oyster trade, has been confirmed as invasive in 17 of the 66 countries where it is cultured. The single study of its status in South Africa reported populations in six South Coast estuaries in 2001, dropping to three sites in 2003. We resurveyed these estuaries, visited others in the Eastern and Western Cape provinces, and sampled oyster tissue for molecular analyses of population structure. Pacific oysters have disappeared from Knysna and, following our collections, possibly also from the GouKou Estuary. Between 2003 and 2012, the Breede Estuary population decreased by 87%, from an estimated 184 206 to 23 760 individuals. Within this estuary, oysters 12 km upriver had denser shells and higher body condition indices than did those within 1.4 km of the river mouth, presumably reflecting higher availability of suspended organic matter. However, low salinity over most of the species' range in the estuary probably inhibits recruitment. New populations of Pacific oysters in the Swartkops and Kaaimans estuaries urgently require monitoring and eradication. Haplotype (h) and nucleotide (π) diversities across all oyster populations sampled (h = 0.2300 [SD 0.0595], π = 0.0006 [SD 0.0007]) were lower than those of co-occurring indigenous Cape rock oysters Striostrea margaritacea from the GouKou and Breede estuaries (h = 0.9076 [SD 0.0386], π = 0.00589 [SD 0.00347]). Pacific oysters either have been introduced to South African estuaries infrequently, or have experienced genetic bottlenecks following river floods or human exploitation, or both. Populations growing outside culture infrastructure are restricted to estuaries in the Eastern and Western Cape provinces, with no evidence of occurrence in fully marine shelf environments. Given the species' considerable socio-economic importance, estuarine and coastal surveillance coupled with aquaculture zoning are required to integrate biodiversity and food security considerations.