First Toxicity Report of Tetrodotoxin and 5,6,11-TrideoxyTTX in the Trumpet Shell Charonia lampas lampas in Europe

Abstract

Tetrodotoxin (TTX) is one of the most potent toxins already isolated, which occurs in a wide variety of animals. In this work, the occurrence of TTX and analogues was examined using mass spectrometry, confocal microscopy, liquid chromatography-mass spectrometry (LC-MS), and mouse bioassay in a trumpet shell (Charonia lampas lampas) and in the fluids of a patient poisoned by consuming this shell. Retention time data in the LC-MS system within the enhanced mass spectrum (EMS) mode indicated the presence of TTX and the analogue 5,6,11-trideoxyTTX; the enhanced product ion (EPI) mode confirmed the existence of both toxins with the formation of characteristic daughter ions from the fragment pattern of each molecule. TTX and 5,6,11-trideoxyTTX were only detected in the digestive gland of the trumpet shell and also in the urine and serum of the patient. The concentration of 5,6,11-trideoxyTTX checked in the samples by LC-MS was 3 times higher than TTX. However, the results obtained by mouse bioassay showed that the analogue is much less toxic than TTX. In vitro toxicity was checked using cerebellar cells; in these experiments the trumpet shell sample showed high toxicity, but the level was lower than in vivo results probably due to some competition between analogues. This paper shows for first time the presence and toxicity of TTX and 5,6,11-trideoxyTTX in a trumpet shell collected in the European coasts. The LC-MS method is a useful tool to confirm the presence of TTX and the further identification of TTX analogues.

New saxitoxin analogues in the marine environment: developments in toxin chemistry, detection and biotransformation during the 2000s

“…Another situation occurred recently in southern Europe, where a serious human poisoning after eating whelks (Charonia lampas) was initially reported as a PSP outbreak in the EU Rapid Alert System for Food and Feed (RASFF Alert Notification: 2007.0752) after testing meal remnants by MBA. Chemical analysis of meal remnants attributed instead this poisoning to the presence of tetrodotoxin analogues, although the exact capture location of the whelks was questionable (Rodríguez et al 2008). Although STX has been reported to occur in several gastropods (Deeds et al 2008), chemical methods are very important tools in order to correctly discriminate between STXs and tetrodotoxins and help to understand the correct source of the contamination.…”

Toxins of Pufferfish—Distribution, Accumulation Mechanism, and Physiologic Functions

“…As described later, a poisoning due to the similar marine snail "kinshibai" Nassarius glans also occurred in Kyushu, Japan, in 2007, respectively (Taniyama et al 2009a. In Spain, the European carnivorous trumpet shell Charonia lampas lampus caused human TTX intoxication in the same year (2007) (Noguchi et al 2011a;Radriguez et al 2008). In New Zealand, 15 dogs were suddenly poisoned at the beaches adjacent to Hauraki Gulf in 2009, all exhibiting similar symptoms, and 5 of them died.…”

Study of the matrix effects of tetrodotoxin and its content in cooked seafood by liquid chromatography with triple quadrupole mass spectrometry

“…TTX is found in a number of marine species, including species of pufferfish [3][4][5][6], gastropods (e.g. nassarius [7][8][9][10], trumpet shell [11], and sea slug [12,13]), xanthid crabs [14,15], red-spotted newts [16,17], and anuran family brachycephalidae [18]. TTX is primarily produced by bacteria, such as actinomyces, aeromonas, alteromonas, bacillus, pseudomonas, vibrio, shewanella algae, and shewanella putrefaciens [19].…”

“…The experimental methods used for the analysis of TTX include LC with a fluorescent detector (FLD) and postcolumn derivatization [23][24][25][26], and LC-MS/MS [3,11,13,[27][28][29][30]. TTX analogs can also be simultaneously screened by both techniques [18,31].…”