Many pufferfish of the family Tetraodontidae possess a potent neurotoxin, tetrodotoxin (TTX). In marine pufferfish species, toxicity is generally high in the liver and ovary, whereas in brackish water and freshwater species, toxicity is higher in the skin. In 1964, the toxin of the California newt was identified as TTX as well, and since then TTX has been detected in a variety of other organisms. TTX is produced primarily by marine bacteria, and pufferfish accumulate TTX via the food chain that begins with these bacteria. Consequently, pufferfish become non-toxic when they are fed TTX-free diets in an environment in which the invasion of TTX-bearing organisms is completely shut off. Although some researchers claim that the TTX of amphibians is endogenous, we believe that it also has an exogenous origin, i.e., from organisms consumed as food. TTX-bearing animals are equipped with a high tolerance to TTX, and thus retain or accumulate TTX possibly as a biologic defense substance. There have been many cases of human intoxication due to the ingestion of TTX-bearing pufferfish, mainly in Japan, China, and Taiwan, and several victims have died. Several cases of TTX intoxication due to the ingestion of small gastropods, including some lethal cases, were recently reported in China and Taiwan, revealing a serious public health issue.

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Tetrodotoxin – Distribution and Accumulation in Aquatic Organisms, and Cases of Human Intoxication

Tetrodotoxin (TTX) is a naturally occurring toxin that has been responsible for human intoxications and fatalities. Its usual route of toxicity is via the ingestion of contaminated puffer fish which are a culinary delicacy, especially in Japan. TTX was believed to be confined to regions of South East Asia, but recent studies have demonstrated that the toxin has spread to regions in the Pacific and the Mediterranean. There is no known antidote to TTX which is a powerful sodium channel inhibitor. This review aims to collect pertinent information available to date on TTX and its analogues with a special emphasis on the structure, aetiology, distribution, effects and the analytical methods employed for its detection.

Tetrodotoxin: Chemistry, Toxicity, Source, Distribution and Detection

Four strains of tetrodotoxin-producing bacteria isolated from a red alga and from pufferfish were characterized. Two of these strains are members of the genus Listonella MacDonell and Colwell. The phenotypic characteristics, guanine-plus-cytosine contents, and base sequences of the 16S rRNAs of these organisms indicated that they are members of Listonella pelagia (Vibrio pelagius) biovar II. The other two strains are members of the genus Alteromonas Baumann et al. and the genus Shewanella MacDonell and Colwell. These two strains are mutually distinct and distinct from the previously described Alteromonas and Shewanella species and therefore are placed in new species. The names Shewanella alga and Alteromonas tetraodonis are proposed for these organisms; the type strains are strains OK-1 and GFC, respectively.

Taxonomy of four marine bacterial strains that produce tetrodotoxin.

On the origins and biosynthesis of tetrodotoxin

TTX accumulation in pufferfish.

A number of type strains of marine bacteria, including members of the family Vibrionaceae, were cultured and examined for tetrodotoxin productivity by high-performance liquid chromatography and gas chromatography-mass spectrometry. Most of the Vibrionaceae strains produced tetrodotoxin, anhydrotetrodotoxin, or both.

Marine bacteria which produce tetrodotoxin.

The occurrence of tetrodotoxin (TTX) in marine sinking particles was investigated. Sinking particles were collected in 1991 using sediment traps in the coastal area of Aburatsubo Inlet, Japan. TTX and related substances were analyzed by tissue culture bioassay, high performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS). TTX and related substances were detected from six samples. The concentrations were estimated to be ca. 200 to 1000 ng g-1 by the HPLC analyses. To our knowledge, this is the first report on the occurrence of TTX in particles in an aquatic environment. The present results indicate that sinking particles are one of the sources of TTX in the marine environment and that these particles play a role in the toxification of marine organisms.

Tetrodotoxin in sinking particles from coastal waters

The occurrence of tetrodotoxin (TTX) in sediment from Lake Suwa, Japan, was confirmed by a tissue culture assay, high-performance liquid chromatography, and gas chromatography-mass spectrometry. Altogether, 17 TTX-producing bacteria belonging to five genera were isolated from Lake Suwa and Pond Inokasira. Our results indicate that TTX and TTX-producing bacteria occur even in freshwater environments.

Presence of tetrodotoxin and tetrodotoxin-producing bacteria in freshwater sediments.

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Marine bacteria which produce tetrodotoxin.

Tetrodotoxin in sinking particles from coastal waters

Presence of tetrodotoxin and tetrodotoxin-producing bacteria in freshwater sediments.