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In 2012, Tetrodotoxin (TTX) was identified in mussels and linked to the presence of  Prorocentrum minimum (P. minimum ) in Greece. The connexion between TTX and  P. minimum  was further studied in this paper. First, the presence of TTX-producer bacteria,  Vibrio  and  Pseudomonas  spp, was confirmed in Greek mussels. In addition these samples showed high activity as inhibitors of sodium currents (I Na ).  P. minimum  was before associated with neurotoxic symptoms, however, the nature and structure of toxins produced by this dinoflagellate remains unknown. Three  P. minimum  strains, ccmp1529, ccmp2811 and ccmp2956, growing in different conditions of temperature, salinity and light were used to study the production of toxic compounds. Electrophysiological assays showed no effect of ccmp2811 strain on I Na , while ccmp1529 and ccmp2956 strains were able to significantly reduce I Na  in the same way as TTX. In these samples two new compounds,  m/z  265 and  m/z  308, were identified and characterized by liquid chromatography tandem high-resolution mass spectrometry. Besides, two TTX-related bacteria,  Roseobacter  and  Vibrio  sp, were observed. These results show for the first time that  P. minimum  produce TTX-like compounds with a similar ion pattern and C9-base to TTX analogues and with the same effect on I Na .

The association of bacterial C9-based TTX-like compounds with Prorocentrum minimum opens new uncertainties about shellfish seafood safety