This study investigates the prevalence of Vibrio spp. in seafood from supermarkets and fish markets in Berlin, Germany. A total of 306 seafood samples, including shrimp and mussels, were bought from supermarkets between March 2023 and January 2024. Samples were analysed using the ISO standard method and multiplex PCR to identify V. parahaemolyticus, V. alginolyticus, V. cholerae and V. vulnificus. The results indicated an overall Vibrio spp. prevalence of 56%. Among the positive samples, the most prevalent species found was V. parahaemolyticus (58%), followed by V. alginolyticus (42%), V. cholerae non-O1/non-O139 (25%), and V. vulnificus (4%). Samples obtained from supermarkets exhibited a lower prevalence (50%) than those received from fish markets (91%). Virulence genes such as ctxA, tdh, or trh were not detected in the respective Vibrio species. Nevertheless, the high prevalence underscores the need and urgency of continuous seafood surveillance.