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• W2035353931 abstract "Tetrodotoxin (TTX) was accumulated at high concentrations in sediment samples from 2 inner water stations and 1 deep-sea station in the western Pacific off the coast of Japan. TTX was identified using tissue culture bioassay and HPLC. It is suggested that bacteria inhabiting the sediment are responsible for the production of toxin. The widespread occurrence of TTX in marine sediments is predicted; such large concentrations of TTX may affect animals inhabiting these sediments. Tetrodotoxin (TTX), known as 'puffer fish toxin', is a potent neurotoxin which blocks the sodium channel of neuronal cells. It has recently been established that this toxin is present in diverse groups of animals, especially those of marine environments (Mosher & Fuhrman 1984, Thuesen et al. 1988). Although mechanisms of TTX accumulation are still obscure, the widespread occurrence of marine bacteria which produce tetrodotoxin (Simidu et al. 1987) suggests that bacteria are responsible for the production of TTX in natural environments. Since marine sediments generally contain large bacterial populations, we investigated the presence of TTX or related sodium channel blocking agents in these sediments and consider the possible accumulation of TTX in the environment. Sediment samples, including one from 4033 m depth, were analyzed by a newly developed tissue culture assay method (Kogure et al. 1988) and by high performance liquid chromatography (HPLC). Sediment samples were taken using a Phleger corer from 2 stations in Tokyo Bay (Stns A and B), which is highly polluted, and 1 station in an open ocean deep area (Stn C), during the KT-87-17 cruise of the RV 'Tansei Maru', Ocean Research Institute, University of Tokyo, in November 1987 (Fig. 1). Sediment samples were separated into layers on board immediately after collection. A portion of them were used to enumerate heterotrophic bacteria, while the rest were kept at -20 C until toxin extraction in the laboratory. Bacterial colonies appearing on ZoBell's 2216E agar plates were counted after 3 wk incubation at 20°C. For the extraction of toxins, sediment samples were thawed, mixed with 10 times their volume of 0.1 % acetic acid, and boiled for 20 min. After centrifugation a t 25 000 X g, the supernatant was freeze-dried and reconstituted in a small amount of distilled water. Samples for HPLC analysis were further purified using SEP-PAK CI8 cartridges (Waters Associates). Tissue culture bioassay (Kogure et al. 1988) was used for the detection and quantitative measurement of sodium channel blocking toxin in the samples. In brief, the alkaloid toxin veratridine causes sodium influx in the mouse neuroblastoma cell line Neuro 2A (ATCC, CCL 131), when the function of Naf-K+ATPase is inhi1 . , ol~eci by uudbdi~ l . This causes celiuiar sweiling and subsequent death; TTX and similar sodium channel blocking agents counteract this effect and enable cells to continue growing. The ratio of living to dead cells can be used to estimate the concentration of toxin in" @default.
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• W2035353931 date "1988-01-01" @default.
• W2035353931 modified "2023-09-27" @default.
• W2035353931 title "Accumulation of tetrodotoxin in marine sediment" @default.
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• W2035353931 doi "https://doi.org/10.3354/meps045303" @default.
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