Search

In recent decades, more than 130 potentially toxic metabolites originating from dinoflagellate species belonging to the genus Karenia or metabolized by marine organisms have been described. These metabolites include the well-known and large group of brevetoxins (BTXs), responsible for foodborne neurotoxic shellfish poisoning (NSP) and airborne respiratory symptoms in humans. Karenia spp. also produce brevenal, brevisamide and metabolites belonging to the hemi-brevetoxin, brevisin, tamulamide, gymnocin, gymnodimine, brevisulcenal and brevisulcatic acid groups. In this review, we summarize the available knowledge in the literature since 1977 on these various identified metabolites, whether they are produced directly by the producer organisms or biotransformed in marine organisms. Their structures and physicochemical properties are presented and discussed. Among future avenues of research, we highlight the need for more toxin occurrence data with analytical techniques, which can specifically determine the analogs present in samples. New metabolites have yet to be fully described, especially the groups of metabolites discovered in the last two decades (e.g tamulamides). Lastly, this work clarifies the different nomenclatures used in the literature and should help to harmonize practices in the future.

En France, de nombreuses toxines non réglementées sont suivies dans le cadre du réseau de veille d’émergence des biotoxines marines dans les coquillages (EMERGTOX) dont le comité de pilotage est présidé par la DGAL et inclut la DGS, la Direction des pêches maritimes et del'aquaculture (DPMA), Santé publique France, l’Anses, l’Ifremer et la Direction de l’eau et de la biodiversité (DEB).Les brévétoxines sont incluses dans ce réseau depuis janvier 2018.Elles ont été mises en évidence pour la première fois en France en novembre 2018 dans des moules en Corse (117 μg/kg de glande digestive). Le prélèvement d’eau associé pour le suivi du phytoplancton a montré la présence de Karenia spp. (genre de microalgues dont certaines espèces produisent des brévétoxines). Il n’existe pas de limite maximale pour ces toxines dans le règlement (CE) n°853/2004. Les USA, l’Australie/Nouvelle-Zélande et le Mexique appliquent un seuil à 800 μg d'équivalent BTX-2/kg de chair de coquillages. Dans la norme CODEXSTAN 292–2008 (rev.2015) du Codex alimentarius, la limite maximale pour les brévétoxines est de 200 unités souris ou équivalent par kg de chair de mollusque vivant.Dans ce contexte, il a été demandé à l’Anses de fournir des éléments de réponse aux questions suivantes :Question 1. Quelles sont les données toxicologiques disponibles pour les brévétoxines ?Permettent-elles à l’Anses de proposer des valeurs toxicologiques de référence aiguë et chronique par voie orale ?Question 2. Sur la base des données toxicologiques identifiées par l’Anses, est-il possible de proposer une valeur guide dans les coquillages au-delà de laquelle des investigations complémentaires seraient demandées dans le cadre du réseau EMERGTOX ?Question 3. Quelles modalités de suivi des brévétoxines dans le milieu marin pourraient être recommandées dans le cadre du réseau EMERGTOX ? Quelles investigations complémentaires seraient à réaliser en cas de dépassement de la valeur guide ?Question 4. Compte tenu des données toxicologiques et au vu du contexte, existe-t-il une préoccupation de santé publique aux niveaux de contamination identifiés dans certaines zones marines françaises liée à la consommation de coquillages ?Question 5. Existe-t-il une préoccupation sanitaire liée à l’exposition aux brévétoxines par un contact direct avec de l’eau contaminée notamment en cas de baignade ou de pratique d’activités nautiques ou en cas d’exposition par inhalation à des embruns ?

Nontarget data acquisition for target analysis (nDATA) workflows using liquid chromatography-high-resolution accurate mass (LC-HRAM) spectrometry, spectral screening software, and a compound database have generated interest because of their potential for screening of pesticides in foods. However, these procedures and particularly the instrument processing software need to be thoroughly evaluated before implementation in routine analysis. In this work, 25 laboratories participated in a collaborative study to evaluate an nDATA workflow on high moisture produce (apple, banana, broccoli, carrot, grape, lettuce, orange, potato, strawberry, and tomato). Samples were extracted in each laboratory by quick, easy, cheap, effective, rugged, and safe (QuEChERS), and data were acquired by ultrahigh-performance liquid chromatography (UHPLC) coupled to a high-resolution quadrupole Orbitrap (QOrbitrap) or quadrupole time-of-flight (QTOF) mass spectrometer operating in full-scan mass spectrometry (MS) data-independent tandem mass spectrometry (LC-FS MS/DIA MS/MS) acquisition mode. The nDATA workflow was evaluated using a restricted compound database with 51 pesticides and vendor processing software. Pesticide identifications were determined by retention time (tR, ±0.5 min relative to the reference retention times used in the compound database) and mass errors (δM) of the precursor (RTP, δM ≤ ±5 ppm) and product ions (RTPI, δM ≤ ±10 ppm). The elution profiles of all 51 pesticides were within ±0.5 min among 24 of the participating laboratories. Successful screening was determined by false positive and false negative rates of

In 2015, tetrodotoxins (TTXs) were considered a potential threat in Europe since several studies had shown the presence of these toxins in European bivalve molluscs. In this study, we investigated the occurrence of TTXs in 127 bivalve samples (mussels and oysters) and in 66 gastropod samples (whelks) collected all along the French mainland coasts in 2017 and 2018. Analyses were carried out after optimization and in-house validation of a performing hydrophilic interaction liquid chromatography associated with tandem mass spectrometry (HILIC-MS/MS) method. The concentration set by European Food Safety Authority (EFSA) not expected to result in adverse effects (44 &micro, g TTX equivalent/kg) was never exceeded, but TTX was detected in three mussel samples and one whelk sample (1.7&ndash, 11.2 &micro, g/kg). The tissue distribution of TTX in this whelk sample showed higher concentrations in the digestive gland, stomach and gonads (7.4 &micro, g TTX/kg) than in the rest of the whelk tissues (below the limit of detection of 1.7 &micro, g TTX/kg). This is the first study to report the detection of TTX in French molluscs.

International audience; Pinnatoxins (PnTXs) are a group of emerging marine biotoxins produced by the benthic dinoflagellate Vulcanodinium rugosum, currently not regulated in Europe or in any other country in the world. In France, PnTXs were detected for the first time in 2011, in mussels from the Ingril lagoon (South of France, Mediterranean coast). Since then, analyses carried out in mussels from this lagoon have shown high concentrations of PnTXs for several months each year. PnTXs have also been detected, to a lesser extent, in mussels from other Mediterranean lagoons and on the Atlantic and Corsican coasts. In the French data, the main analog is PnTX G (low levels of PnTX A are also present in some samples). No cases of PnTXs poisoning in humans have been reported so far in France or anywhere else in the world. In mice, PnTXs induce acute neurotoxic effects, within a few minutes after oral administration. Clinical signs of toxicity include decreased mobility, paralysis of the hind legs, tremors, jumps and breathing difficulties leading to death by respiratory arrest at high doses. The French agency for food safety (ANSES) recently conducted a review of the state of knowledge related to PnTXs and V. rugosum. Based on (i) the clinical signs of toxicity in mice, (ii) the mode of action of PnTXs as nicotinic acetylcholine receptor competitive antagonists and (iii) knowledge on drugs and natural toxins with PnTX-related pharmacology, potential human symptoms have been extrapolated and proposed. In this work, a provisional acute benchmark value for PnTX G of 0.13 μg/kg bw per day has been derived from an oral acute toxicity study in mice. Based on this value and a large shellfish meat portion size of 400g, a concentration lower than 23 μg PnTX G/kg shellfish meat is not expected to result in adverse effects in humans. ANSES recommends taking into account PnTXs in the French official monitoring program for shellfish production and identified data gaps to refine health risk assessment.

International audience; Consumers generally considered organic products to be healthier and safer but data regarding the contamination of organic products are scarce. This study evaluated the impact of the farming system on the levels of ochratoxin A (OTA) in the tissues of French pigs (muscle and liver) reared following three different types of production (organic, Label Rouge and conventional). Because OTA is present at trace levels in animal products, a sensitive ultra-high performance liquid chromatography-tandem mass spectrometry method using stable isotope dilution assay was developed and validated. OTA was detected or quantified (LOQ of 0.10 mu g kg(-1)) in 67% (n = 47) of the 70 pig liver samples analysed, with concentrations ranging from < 0.10 to 3.65 mu g kg(-1). The maximum concentration was found in a sample from organic production but there were no significant differences in the content of OTA between farming systems. OTA was above the LOQ in four out of 25 samples of the pork muscles. A good agreement was found between OTA levels in muscle and liver (liver concentration = 2.9 x OTA muscle concentration, r = 0.981).

International audience; This study assessed the transfer of fumonisins B1 and B2 (FBs), zearalenone (ZEA) and zearalenone metabolites alpha-zearalanol (alpha-ZAL) and alpha-zearalenol (alpha-ZEL) to poultry tissues. Two experimental groups of 9 male chickens each were exposed for 8 weeks to a contaminated diet (FB group: 12.7 mg FB1 + FB2 kg(-1), ZEA group: 0.40 mg ZEA kg(-1)). To measure the carry-over from feed to animal tissues (liver and muscle), a sensitive and accurate SIDA-UHPLC-MS/MS method was developed and validated. For all mycotoxins, the limit of detection (LOD) was 0.3 mu g kg(-1) and the limit of quantification (LOQ) was 1.0 mu g kg(-1). Recoveries ranged from 92% to 107% and the intermediate precision coefficients of variation (CVIP) were between 4.3% and 13%. ZEA and alpha-ZAL were not detected in livers, whereas alpha-ZEL was detected in five out of eight samples at levels between LOD and LOQ. FBs were detected and quantified in the livers of all animals exposed to the contaminated diet (mean 30.3 mu g FB1 kg(-1) and 2.3 mu g FB2 kg(-1)). A significant correlation between the FB1 and the FB2 contents in the liver was demonstrated and FB carry-over factors (CFs) from feed to liver were determined (CFFB1: 0.003 and CFFB2: 0.001). Filet muscles from the same animals were also analysed. FB1 was quantified at trace levels in eight samples out of nine (mean 2.0 mu g kg(-1)) and was only detected in the remaining sample. FB2 was detected in only one muscle sample.

From January 2011 to March 2018, 26 patients aged from 20 to 80 years old reported being sick in France after eating sea figs of the genus Microcosmus. The patients had symptoms evoking a cerebellar syndrome: blurred or double vision, ataxia and dizziness, asthenia, headache, muscle cramps, paresthesia and digestive disorders (nausea, vomiting and diarrhea). Three of the 18 food poisoning events recorded by the Poison Control Center in Marseille and involving four patients were further investigated as the meal leftovers were collected and analyzed. A previous study ruled out the presence of the regulated lipophilic marine toxins after high-resolution mass spectrometry, but further analyses were required to look for hydrophilic cyanotoxins. The sea fig leftovers from food poisoning case Numbers 1 (January 2011), 6 (December 2012) and 17 (March 2018) of this published case series were analyzed by hydrophilic interaction liquid chromatography coupled to low- and high-resolution mass spectrometry to investigate the presence of hydrophilic cyanotoxins. The sea fig samples showed anatoxin-a (ATX-a) concentrations ranging from 193.7 to 1240.2 &micro, g/kg. The sea fig control sample analyzed was also contaminated with ATX-a but in a much smaller concentration (22.5 &micro, g/kg). To the best of our knowledge, this is the first report of human food poisoning involving ATX-a as the possible causative toxin where the cyanotoxin could be unequivocally identified.

International audience; When considering the geographical expansion of marine toxins, the emergence of new toxins and the associated risk for human health, there is urgent need for versatile and efficient analytical methods that are able to detect a range, as wide as possible, of known or emerging toxins. Current detection methods for marine toxins rely on a priori defined target lists of toxins and are generally inappropriate for the detection and identification of emerging compounds. The authors describe the implementation of a recent approach for the non-targeted analysis of marine toxins in shellfish with a focus on a comprehensive workflow for the acquisition and treatment of the data generated after liquid chromatography coupled with high resolution mass spectrometry (LC-HRMS) analysis. First, the study was carried out in targeted mode to assess the performance of the method for known toxins with an extended range of polarities, including lipophilic toxins (okadaic acid, dinophysistoxins, azaspiracids, pectenotoxins, yessotoxins, cyclic imines, brevetoxins) and domoic acid. The targeted method, assessed for 14 toxins, shows good performance both in mussel and oyster extracts. The non-target potential of the method was then challenged via suspects and without a priori screening by blind analyzing mussel and oyster samples spiked with marine toxins. The data processing was optimized and successfully identified the toxins that were spiked in the blind samples. Key Contribution: (1) Efficient liquid chromatography coupled with high resolution mass spectrometry (LC-HRMS) targeted quantitative method developed and characterized for the analysis of many marine toxins of different polarities, including lipophilic toxins and domoic acid. (2) Optimized strategy for non-targeted analysis of marine toxins, including suspect and without a priori screening. (3) Combination of targeted and non-targeted strategies allowing for good characterization of analyzed shellfish samples.

International audience; The chemical contamination levels of both conventional and organic meats were assessed. The objective was to provide occurrence data in a context of chronic exposure. Environmental contaminants (17 polychlorinated dibenzodioxins/dibenzofurans, 18 polychlorinated biphenyls (PCBs), 3 hexabromocyclododecane (HBCD) isomers, 6 mycotoxins, 6 inorganic compounds) together with chemical residues arising from production inputs (75 antimicrobials, 10 coccidiostats and 121 pesticides) have been selected as relevant compounds. A dedicated sampling strategy, representative of the French production allowed quantification of a large sample set (n=266) including both conventional (n=139) and organic (n=127) raw meat from three animal species (bovine, porcine, poultry). While contamination levels below regulatory limits were measured in all the samples, significant differences were observed between both species and types of farming. Several environmental contaminants (Dioxins, PCBs, HBCD, Zn, Cu, Cd, Pb, As) were measured at significantly higher levels in organic samples.