Per- and polyfluorinated alkyl substances (PFAS) are highly persistent chemicals, which pose a potential risk for aquatic wildlife due to their bioaccumulative behaviour and toxicological effects. Although the distribution of PFAS in marine environments has been studied worldwide, little is known on the contamination of PFAS in the southern North Sea. In the present study, the bioaccumulation and trophic transfer of Perfluoroalkyl acids (PFAAs) was studied in liver and muscle tissue of seven fish species and in whole-body tissue of two crustacean species, collected at 10 sites in the Belgian North Sea. Furthermore, the human and ecological health risks were examined. Overall, perfluorooctane sulfonate (PFOS) was predominant in all matrices and other long-chain PFAS were frequently detected. Mean PFOS concentrations ranged from <LOQ to 107 ng/g (ww) in fish liver, from <LOQ to 24 ng/g ww in fish muscle and from 0.29 to 5.6 ng/g ww in crustaceans. Elevated perfluorotridecanoic acid (PFTrDA) concentrations were detected in fish liver from the estuarine and coastal region (<LOQ-116 ng/g ww), indicating a specific point source of this compound. Based on stable isotope analysis, no distinctive trophic transfer patterns of PFAS could be identified which implies that the bioconcentration of PFAS from the surrounding abiotic environment is most likely dominating over the biomagnification in the studied biota. The consumption of commercially important species such as the brown shrimp (Crangon crangon), plaice (Pleuronecta platessa), sole (Solea solea) and whiting (Merlangus merlangus) might pose potential health risks if it exceeds 17 g/day, 18 g/day, 26 g/day and 43 g/day respectively. Most PFOS measurements did not exceed the QS_biota,hh of 9.1 ng/g ww, however, the benchmark of 33 ng/g ww targeting the protection of wildlife from secondary poisoning was exceeded for 43% and 28% of the samples in plaice and sole.