Pierre Masselin

Short-time cytotoxicity of mussel extracts: A new bioassay for okadaic acid detection

Okadaic acid (OA), the main toxin responsible for diarrhoeic shellfish poisoning (DSP) has high cytotoxicity for KB cell cultures (apparent after 3 hr of contact), facilitating rapid detection in contaminated mussels. We developed a method to determine the minimal active concentration (MAC) based on direct microscopic study of toxin-induced changes in cell morphology. A high correlation was found between the MAC of tested extracts and corresponding OA concentrations in mussel hepatopancreas as measured by high performance liquid chromatography. This technique is rapid and reproducible and does not require the use of living animals.

High performance liquid chromatography analysis of diarrhetic toxins inDinophysis spp. from the French coast

Diarrhetic shellfish poisoning has been a recurrent problem along the Brittany coast (France) since 1983. Okadaic acid (O.A.), the main toxin detected in mussels, is generally associated with the presence ofDinophysis cells in sea water. We report here the results of okadaic acid analyses by high performance liquid chromatography, on planktonic samples collected during the summer of 1991.D. sacculus, the majorDinophysis species, throughout this period, showed low okadaic acid content in raw extract, whereas toxic levels were 3- to 12-fold higher in sorted samples than in raw ones. A maximum O.A. concentration of 29.6 pg cell-1 was found inD. sacculus/D. cf. acuminata extract. Similarly, higher O.A. levels were noted in raw samples when these two species were associated. Generally speaking, variations in raw samples were similar to those in sorted samples. Nevertheless, whenD. rotundata reached a concentration equal to that ofD. sacculus in sorted samples, the O.A. level was lower.

Paralytic shellfish poison accumulation yields and feeding time activity in the Pacific oyster (Crassostrea gigas) and king scallop (Pecten maximus)

Pacific oysters and king scallops placed individually in a recirculating flume were fed for 2 weeks with a constant concentration (120 cell ml ‐1 ) of a toxic strain of Alexandrium minutum. Fluorescence at the outlet of each experimental unit was measured continuously, and biodeposits were recovered twice daily to evaluate feeding time activity (FTA) and rates of organic filtration (OFR), ingestion (OIR) and organic absorption (OAR). Ion-pairing high performance liquid chromatography (IP-HPLC) was performed concurrently to quantify paralytic phycotoxins both (i) individually in shellfish at the end of the contamination period and (ii) in A. minutum cultures to estimate cellular toxin concentration. These data allowed comparison of the actual tissue toxin concentration (TOX) with the theoretical toxin accumulation rate (TAR) calculated from the linear relations between OAR, cell number, fluorescence and cell toxicity. The results show high FTA/TAR and FTA/TOX correlations (R 2 = 0.78) for both oysters and scallops. The TAR/TOX relation, though not yet clearly defined, suggests the minimum quantity of absorbed toxin at which the accumulation process is induced.