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Non-toxic scallops Patinopecten yessoensis were reared for 5 days with the toxic dinoflagellate Dinophysis fortii sampled from the sea. The total cell number ingested by five scallops during the feeding experiment was approximately 682 x 10(3). Okadaic acid (OA), dinophysistoxin-1 (DTX1) and esterified toxins of OA and DTX1 in D. fortii and scallop extracts were determined by liquid chromatography fluorescence detection (LC-FLD) as their 9-anthryldiazomethane (ADAM) derivatives. Only DTX1 was detected in D. fortii extracts used for the feeding experiment. The content of esterified DTXI in scallops fed on D. fortii was significantly higher than that of DTX1. Identification of esterified DTX1, 7-O-palmitoyl DTX1 (7-O-16:0 DTX1), in D. fortii and scallop extracts was carried out by LC equipped with an atmospheric pressure electrospray ionization-mass spectrometry (ESI-MS). Only extracts of scallops fed on D. fortii yielded a mass spectrum exhibiting abundant [M-H]- at m/z 1055 corresponding to 7-O-16:0 DTX1. These findings are the first direct evidence of the transformation of DTX1 to 7-O-acyl DTX1 in scallop tissues.

Solid-phase extraction of pectenotoxin-2 (PTX2) and pectenotoxin-6 (PTX6) from seawater samples using a nonpolar cartridge column Sep-Pak C18 was investigated for determination of PTXs in toxic phytoplankton. PTX2 and PTX6 were almost completely recovered from the seawater samples. Determinations of PTX2 from the toxic phytoplankton and scallops were carried out by liquid chromatography (LC) on UV trace equipped with an atmospheric pressure electrospray ionization mass spectrometry (ESI-MS). PTX2 obtained from the toxic phytoplankton and scallops yielded a mass spectra exhibiting abundant [M+H]+, [M+NH4]+ and [M+Na]+ indicating that ESI-MS is useful for the identification of PTX2. Determinations of PTX6 from the toxic phytoplankton and scallops were carried out by LC fluorescence detection. A significantly higher content of PTX6 compared to PTX2 in scallops was observed, suggesting that a transformation of PTX2 to PTX6 in tissues of the scallops occurs rapidly.

Determination of free fatty acids (FFA) in hepatopancreas of mussel, Mytilus coruscus, contaminated by diarrhetic shellfish poisoning (DSP) toxins, was carried out by high-performance liquid chromatography-fluorometry as their 9-anthryldiazomethane derivatives. The FFA in the hepatopancreas of mussels during May 30-June 14 were 443-666 micrograms/g. A marked increase in FFA content was observed on June 20 (2197 micrograms/g hepatopancreas) and June 27 (6322 micrograms/g hepatopancreas). The proportion of polyunsaturated fatty acids (PUFA), including 20:4n-6, 20:5n-3, and 22:6n-3, in mussel FFA increased during the experimental period. Diatoms almost completely dominated the phytoplankton community at the experimental site, suggesting that diatoms were the origin of the PUFA accumulated in mussels. The toxicity of FFA in mussel hepatopancreas on June 27 was sufficiently high as to interfere with the mouse bioassay by injection of an extract of shellfish, the official method for DSP toxin analysis.

This study clarified the spawning mechanism of female rose bitterling, Rhodeus ocellatus ocellatus, with special emphasis on the physical role of urine. Ovulated females ingested a significantly greater quantity of water by drinking than non-ovulated fish. The body weight of ovulated females increased about 1.3% while it decreased 1.0% in non-ovulated females. Urine volume in the urinary bladder increased rapidly before spawning, synchronized with the spawning cycle. On the other hand, little urine remained in specimens which had completed spawning. Histological observation demonstrated that the oviduct and the ureter joined at the proximal part of the ovipositor. On the basis of these anatomical and physiological results, it is proposed that urine plays the physical role of pushing the ovulated eggs through the elongated ovipositor during spawning.