ATLANTOX
Advanced Test about New toxins appeared in Atlantic Area
Scientific results:
In the ATLANTOX project framework, several samples of shellfish from the Atlantic coast were analyzed. Most of them were positive for the presence of spirolids, tetrodotoxin, or cyanotoxins. Toxic episodes are a major public health problem whose impact is moved to touristic areas and, consequently, there is a reduction on consumption of seafood. Therefore, the HABs cause significant economic losses due to the accumulation of toxins in shellfish.Spirolids are the main group of cyclic imines toxins; their appearance is increasingly around the world. The most representative compound of this group is 13-desmethyl spirolide C (13-desMeC). This project contributed to the elucidation of the mechanism of action of 13-desMeC. Muscarinic acetylcholine receptors were identified as the cellular target of 13-desMeC, these receptors were synthesized and their binding were studied. Information on the identification and characterization of the hazard of these new toxins has been reviewed and presented throughout the ATLANTOX project. The literature on risk assessment of marine biotoxins in the EU was reviewed. It is also concluded that a revision of the present regulation of marine biotoxins in the EU legislation could be considered, as some regulated toxins do not show a risk for EU's population (as yessotoxin) and some non regulated toxins have been shown to be harmful and/or to occur in the EU (as tetrodotoxin, palytoxin, and some cyclic imines) while they are not regulated. In the ATLANTOX project, studies of toxicity and identification of the targets for marine toxins which causes problems in the food security have been developed. Also, fast and alternative methods for the toxin control based in functional antibodies were also developed.
Sensible and specific functional detection methods for spirolides, tetrodotoxins and palitoxins with good limits of quantification were developed. Some of these were validated and therefore, it can be used as official methods in the possible regulation of these toxins. The method for cyclic imine toxins detection is based on the competition of spirolide with the alfa-bungarotoxin for the binding of the acetilcholine receptors enriched by Torpedo marmorata membranes. This assay uses the fluorescence polarization technique and detects spirolides with high especificity, sensibility and good toxin recovery. Using the same Torpedo marmorata membranes, the first direct detection method using fluorescence polarization was developed. A functional method to detect cyclic imines using chemiluminescence, fluorescence and colorimetry was also developed. For tetrodotoxin, a method based on the biosensor technique was developed. The validation was carried out according to European Decision 2002/657/EC guidelines along with the IUPAC (International Union of Pure and Applied Chemistry) and AOAC. Several issues as decision limit, detection limit, recovery, selectivity, repeatability, method robustness were addressed. The effect of the matrix in samples of Charonia lampas from Portugal were checked. For tetrodotoxin, a analytical method using Liquid-chromatography-electrospray ionization tandem-mass spectrometry (LC-ESI-CID-MS/MS) technique was developed. This method has a low detection limit 0.08 µg/g and it was employed to analyze samples of puffer fish (sceleratus Lagocephalus) from Europe coast (Aegean Sea, Greece). Tetrodotoxin and 3 analogs (5,6,11-trideoxyTTX, 11-deoxyTTX y 11-norTTX-6(S)-ol) were found in this samples. For palitoxin, a method based in biosensor technique and another one based in fluorescence polarization technique was developed. Also, a detection and quantification method for palitoxin using phage libraries which contain antibodies was developed. In summary, good detection methods to be used in toxin marine control were developed in the framework of ATLANTOX project.
International scope and diffusion of results
Participation in conferences, like 8th International Conference in Molluscan Shellfish Safety. Charlottetown (Canada), 12th-17th June. Presentation of the first results of methods pre-validation for toxin detection, developed throughout ATLANTOX: Microplate-based cyclic imines detection kit. ASE-UPLC-MS/MS for tetrodoxin. SPR biosensor for tetrodoxin
-Dissemination for attendees to events.
-Updating of project website with new contents
- Updating of internet database on marine biotoxins
The database on marine biotoxins contains now information about the main groups of marine biotoxins, including references of relevant scientific article. A new category containing links to relevant scientific and dissemination articles or other sources of information has been created. General information about marine biotoxins has been disseminated through the webpage and the internet accessible marine biotoxins database.
The project has been publicized among the scientific community, relevant stakeholders and general public.
The partners of ATLANTOX project continue with the exchange of information and working for the PHARMATLANTIC project. PHARMATLANTIC is a project born on necessity to provide involved industries situated in the Atlantic seaboard with research and innovation advances in prevention of mental diseases and cancer already developed by the most specialized centres in this field at EU level.
