Ciguatoxin Research Articles

Tropical fish poisonings in Germany 2012–2017 - What is ciguatera?

Purpose: Ciguatera fish poisoning (CFP) is the most common fish intoxication worldwide caused by ciguatoxin(CTX)-containing tropical reef fish. CFP is dominated by long-lasting neurologic symptoms, whereas gastro-intestinal symptoms may be present in the acute phase, only. In continental Europe, CFP may occur when CTX-contaminated fish is consumed during or after trips to endemic areas, or when it is exported to non-endemic countries. At the subtropical European Canary Islands and Madeira, CFP has been reported since the early 2000s. CTX-producing microalgae have been recently found in the Mediterranean Sea. In Germany, a CFP outbreak occurred in each of the years 2012 to 2017 caused by snappers imported from India, Vietnam and Indonesia. As outbreak investigations showed, German physicians are not aware of CFP, and patients were unsatisfied with medical care. Clinical and epidemiological characteristics of this series of CFP outbreaks are demonstrated here. Channels to report CFP cases and outbreaks to Public Health authorities without an established notification system will be shown. Methods & Materials: Reports by physicians according to §16e of the German Chemicals Act were analyzed in order to retrieve CFPs based on the case definition of the EuroCigua-project. The degree of severity was evaluated on the bases of the Poison Severity Score (PSS). CFP cases were clustered according to the place where the fish was purchased. Results: Between 2012 and 2017, six outbreaks with 65 CFP cases were registered in Germany. All patients suffered from neurologic disorders. Three patients showed severe symptoms like bradycardia <40 min-1 or morphine resistant abdominal cramps. Nearly 20% of the patients were hospitalized. All outbreak cases were caused by snappers from India, Indonesia and Vietnam. Lutjanus bohar was the fish species present in all German CFP outbreaks. Conclusion: Ciguatera caused by imported tropical fish is an emerging disease in Germany. Physicians should acquaint knowledge of CFP symptoms and therapeutic options, and are requested to report CFP to the German Federal Institute for Risk Assessment (BfR). CFP patients should be reminded to provide fish leftovers to local Public Health authorities for laboratory investigations. DRG code T 61.0 should be used when the CFP case definition of the EuroCigua project is applicable.

The Incidence of Marine Toxins and the Associated Seafood Poisoning Episodes in the African Countries of the Indian Ocean and the Red Sea.

The occurrence of Harmful Algal Blooms (HABs) and bacteria can be one of the great threats to public health due to their ability to produce marine toxins (MTs). The most reported MTs include paralytic shellfish toxins (PSTs), amnesic shellfish toxins (ASTs), diarrheic shellfish toxins (DSTs), cyclic imines (CIs), ciguatoxins (CTXs), azaspiracids (AZTs), palytoxin (PlTXs), tetrodotoxins (TTXs) and their analogs, some of them leading to fatal outcomes. MTs have been reported in several marine organisms causing human poisoning incidents since these organisms constitute the food basis of coastal human populations. In African countries of the Indian Ocean and the Red Sea, to date, only South Africa has a specific monitoring program for MTs and some other countries count only with respect to centers of seafood poisoning control. Therefore, the aim of this review is to evaluate the occurrence of MTs and associated poisoning episodes as a contribution to public health and monitoring programs as an MT risk assessment tool for this geographic region.

New Insights into the Occurrence and Toxin Profile of Ciguatoxins in Selvagens Islands (Madeira, Portugal).

Ciguatoxins (CTXs), endemic from tropical and subtropical regions of the Pacific and Indian Ocean and the Caribbean Sea, have caused several human poisonings during the last decade in Europe. Ciguatera fish poisonings (CFP) in Madeira and Canary Islands appear to be particularly related with consumption of fish caught close to Selvagens Islands, a Portuguese natural reserve composed of three small islands that harbor high fish biomass. In this study, fish specimens considered as potential vectors of CTXs were caught in Madeira and Selvagens archipelagos for toxins determination via sensitive liquid chromatography with tandem mass spectrometry detection (LC–MS/MS). CTXs were found in most of the fish samples from Selvagens and none from Madeira. Caribbean ciguatoxin-1 (C-CTX1) was the only toxin congener determined, reaching the highest value of 0.25 µg C-CTX1 kg−1 in a 4.6 kg island grouper (Mycteroperca fusca). This study indicates that a diversity of fish from different trophic levels contains CTXs, Selvagens appear to be one of the most favorable locations for CTXs food web transfer and finally, this study highlights the need of further research based on intensive environmental and biological sampling on these remote islands.

Gambierdiscus jejuensis sp. nov., an epiphytic dinoflagellate from the waters of Jeju Island, Korea, effect of temperature on the growth, and its global distribution

The genus Gambierdiscus produces ciguatera toxins, which has led to extensive studies of this genus. Epiphytic dinoflagellate cells were isolated from coralline macroalgae collected from the coastal waters of Jeju Island, Korea, and two clonal cultures were established. The morphology of these cells was examined using light and scanning electron microscopy, and the sequences of the small subunit (SSU), large subunit (LSU), and internal transcribed spacer (ITS) region of rDNA were determined. The phylogenetic trees based on SSU, LSU (D1-D3), and LSU (D8-D10) rDNA showed the two Korean isolates forming a clade with unidentified strains named Gambierdiscus sp. type 2. This clade showed a clear divergence from the two closest clades containing of the G. caribaeus and G. carpenteri strains. Morphologically, the Korean isolates had an anteroposteriorly compressed cell shape having a smooth theca ornamentation. Growth of the Korean isolates ceased when water temperatures were <20 °C or >30 °C with an optimum at 26 °C. This temperature preference may account for why Gambierdiscus sp. type 2 has only been isolated from the temperate waters of Korea and Japan. However, the two most closely related species G. caribaeus and G. carpenteri have broader temperature ranges tolerating as high as 34–36 °C and are globally distributed. Based on these results, the Korean isolates are proposed as a new species in the genus Gambierdiscus and named Gambierdiscus jejuensis sp. nov. Furthermore, it is suggested that the unidentified strains belonging to the Gambierdiscus sp. type 2 be transferred to the new species, G. jejuensis.

Consumption of barracuda in the Caribbean Sea linked to ciguatera fish poisoning among Filipino seafarers.

IntroductionCiguatera fish poisoning (CFP) is common in tropical and subtropical waters. On 13 November 2015, eight Filipino seafarers from a cargo ship sailing in the Caribbean Sea experienced a range of symptoms after consuming a barracuda. Upon their return to the Philippines, an investigation was conducted to describe the cases.MethodsA case-series was conducted. A CFP case was defined as a previously well individual on the ship who developed at least one gastrointestinal symptom and at least one neurologic manifestation after eating barracuda on 13 November 2015. All cases were admitted to hospital in Manila, Philippines and were interviewed using a standard questionnaire. Urine and serum samples of cases were collected for ciguatoxin (CTX) testing by radiological and receptor-binding assay.ResultsEight of the 25 seafarers on the ship ate the barracuda; all eight met the CFP case definition. The age of cases ranged from 37 to 58 years (median: 47 years) and all were males. Onset of symptoms ranged from 1 to 3 hours (median: 2 hours) from the time of ingestion of the barracuda. All cases experienced gastrointestinal (nausea, vomiting, diarrhoea) and neurologic (temperature allodynia, itchiness) symptoms but no cardiovascular manifestations. Urine and serum specimens of all eight cases showed CTX below the detection limit.DiscussionThe Philippines Epidemiology Bureau recommended that the Philippine Maritime Authority include CTX poisoning and its health risks in seafarers’ training to prevent future cases of CFP. The Event-based Surveillance and Response system will continue to provide a mechanism for the reporting and appropriate management of CFP cases.

Design of experiments for the optimization of electrospray ionization in the LC-MS/MS analysis of ciguatoxins.

Liquid chromatography (LC) coupled to mass spectrometry (MS) is being widely applied as an analytical tool in the field of marine biotoxins both for regulated and for new and emerging compounds. LC-MS/MS recently became the reference method for the control of lipophilic toxins in the European Union, and new methods are being developed and optimized to extend the applicability of this technique to other toxin groups. In this work, conditions for the analysis of ciguatoxins (CTXs) by LC-MS/MS were investigated using standard solutions of CTX1B and CTX3C, which are structurally representative compounds for the rest of the main congeners of Pacific group toxins (P-CTXs). Preliminary studies were carried out for the selection of precursor and product ions used for multiple reaction monitoring. Two transitions based on the chemical structures of CTXs were set up, and mass spectrometer parameters were adjusted for selected reactions monitored. The electrospray ionization source has been carefully optimized through a design of experiments that consisted of a two-level fractional factorial design of resolution IV for the screening of adequate source conditions and of response surface designs for optimization of the main interactions between factors. The statistical approach allowed maximizing the sensitivity on the MS analyzer that provides a good specificity in P-CTX detection, which can be also used for confirmation purposes.

Investigation of ciguatoxins in invasive lionfish from the greater caribbean region: Implications for fishery development.

Lionfish, native to reef ecosystems of the tropical and sub-tropical Indo-Pacific, were introduced to Florida waters in the 1980s, and have spread rapidly throughout the northwestern Atlantic, Caribbean Sea and the Gulf of Mexico. These invasive, carnivorous fish significantly reduce other fish and benthic invertebrate biomass, fish recruitment, and species richness in reef ecosystems. Fisheries resource managers have proposed the establishment of a commercial fishery to reduce lionfish populations and mitigate adverse effects on reef communities. The potential for a commercial fishery for lionfish is the primary reason to identify locations where lionfish accumulate sufficient amounts of ciguatoxin (CTX) to cause ciguatera fish poisoning (CFP), the leading cause of non-bacterial seafood poisoning associated with fish consumption. To address this issue, an initial geographic assessment of CTX toxicity in lionfish from the Caribbean and Gulf of Mexico was conducted. Lionfish samples (n = 293) were collected by spearfishing from 13 locations (74 sampling sites) around the Caribbean and Gulf of Mexico between 2012 and 2015. The highest frequencies of lionfish containing measurable CTX occurred in areas known to be high-risk regions for CFP in the central to eastern Caribbean (e.g., 53% British Virgin Islands and 5% Florida Keys). Though measurable CTX was found in some locations, the majority of the samples (99.3%) contained CTX concentrations below the United States Food and Drug Administration guidance level of 0.1 ppb Caribbean ciguatoxin-1 (C-CTX-1) equivalents (eq.). Only 0.7% of lionfish tested contained more than 0.1 ppb C-CTX-1 eq. As of 2018, there has been one suspected case of CFP from eating lionfish. Given this finding, current risk reduction techniques used to manage CTX accumulating fish are discussed.

Assessment of cytotoxicity in ten strains of Gambierdiscus australes from Macaronesian Islands by neuro-2a cell-based assays

Within the dinoflagellate genus Gambierdiscus several species are well-known producers of ciguatoxins (CTXs) and maitotoxins (MTXs). These compounds are potent marine toxins that accumulate through the food chain, leading to a foodborne disease known as ciguatera when contaminated fish is consumed. Given the threat that the presence of these toxins in seafood may pose to human health and fisheries, there is an evident necessity to assess the potential toxicity of Gambierdiscus sp. in a particular area. Thus, the purpose of this work was to evaluate the production of CTX and MTX of 10 strains of Gambierdiscus australes isolated from the Selvagem Grande Island (Madeira, Portugal) and El Hierro Island (Canary Islands, Spain). The strains were first characterized by light microscopy and species were confirmed by molecular biology, being identified as G. australes. Following the species identification, CTX and MTX production of G. australes extracts was evaluated at the exponential growth phase using neuro-2a cell-based assays. Additionally, the production of MTX was also investigated in two of the G. australes strains collected at the stationary growth phase. Interestingly, 9 out of 10 strains were found to produce CTX-like compounds, ranging from 200 to 697 fg equiv. CTX1B cell−1. None of the G. australes strains showed MTX-like activity at the exponential phase, but MTX production was observed in two strains at the stationary growth phase (227 and 275 pg equiv. MTX cell−1). Therefore, the presence of G. australes strains potentially producing CTX and MTX in these Macaronesian Islands was confirmed.

Experimental evidence of dietary ciguatoxin accumulation in an herbivorous coral reef fish

Ciguatoxins (CTXs) are potent algal toxins that cause widespread ciguatera poisoning and are found ubiquitously in coral reef food webs. Here we developed an environmentally-relevant, experimental model of CTX trophic transfer involving dietary exposure of herbivorous fish to the CTX-producing microalgae Gambierdiscus polynesiensis. Juvenile Naso brevirostris were fed a gel-food embedded with microalgae for 16 weeks (89 cells g−1 fish daily, 0.4 μg CTX3C equiv kg−1 fish). CTXs in muscle tissue were detectable after 2 weeks at levels above the threshold for human intoxication (1.2 ± 0.2 μg CTX3C equiv kg−1). Although tissue CTX concentrations stabilized after 8 weeks (∼3 ± 0.5 μg CTX3C equiv kg−1), muscle toxin burden (total μg CTX in muscle tissue) continued to increase linearly through the end of the experiment (16 weeks). Toxin accumulation was therefore continuous, yet masked by somatic growth dilution. The observed CTX concentrations, accumulation rates, and general absence of behavioural signs of intoxication are consistent with field observations and indicate that this method of dietary exposure may be used to develop predictive models of tissue-specific CTX uptake, metabolism and depuration. Results also imply that slow-growing fish may accumulate higher CTX flesh concentrations than fast-growing fish, which has important implications for global seafood safety.

Highly Sensitive and Practical Fluorescent Sandwich ELISA for Ciguatoxins.

Ciguatera fish poisoning (CFP) caused by the consumption of fish that have accumulated ciguatoxins (CTXs) affects more than 50000 people annually. The spread of CFP causes enormous damage to public health, fishery resources, and the economies of tropical and subtropical endemic regions. The difficulty in avoiding CFP arises from the lack of sensitive and reliable analytical methods for the detection and quantification of CTXs in contaminated fish, along with the normal appearance, smell, and taste of fish contaminated with the causative toxins. Thus, an accurate, sensitive, routine, and portable detection method for CTXs is urgently required. We have successfully developed a highly sensitive fluorescent sandwich ELISA, which can detect, differentiate, and quantify four major CTX congeners (CTX1B, CTX3C, 51-hydroxyCTX3C, and 54-deoxyCTX1B) with a detection limit of less than 1 pg/mL. The ELISA protocol, using one microtiter plate coated with two mAbs (10C9 and 3G8), and ALP-linked 8H4, can detect any of the four CTX congeners in a single operation. CTX1B spiked into fish at the FDA guidance level of 0.01 ppb CTX1B equivalent toxicity in fish from Pacific regions was also proven to be reliably detected by this ELISA. Furthermore, the efficiency of extraction/purification procedures and the matrix effect of contaminants in fish were evaluated in detail, since pretreatment and matrix effects are critical for ELISA analysis.

Tissue Distribution and Elimination of Ciguatoxins in Tridacna maxima (Tridacnidae, Bivalvia) Fed Gambierdiscus polynesiensis.

Ciguatera is a foodborne disease caused by the consumption of seafood contaminated with ciguatoxins (CTXs). Ciguatera-like poisoning events involving giant clams (Tridacna maxima) are reported occasionally from Pacific islands communities. The present study aimed at providing insights into CTXs tissue distribution and detoxification rate in giant clams exposed to toxic cells of Gambierdiscus polynesiensis, in the framework of seafood safety assessment. In a first experiment, three groups of tissue (viscera, flesh and mantle) were dissected from exposed individuals, and analyzed for their toxicity using the neuroblastoma cell-based assay (CBA-N2a) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses. The viscera, flesh, and mantle were shown to retain 65%, 25%, and 10% of the total toxin burden, respectively. All tissues reached levels above the safety limit recommended for human consumption, suggesting that evisceration alone, a practice widely used among local populations, is not enough to ensure seafood safety. In a second experiment, the toxin content in contaminated giant clams was followed at different time points (0, 2, 4, and 6 days post-exposure). Observations suggest that no toxin elimination is visible in T. maxima throughout 6 days of detoxification.

Toxicological Investigations on the Sea Urchin Tripneustes gratilla (Toxopneustidae, Echinoid) from Anaho Bay (Nuku Hiva, French Polynesia): Evidence for the Presence of Pacific Ciguatoxins.

The sea urchin Tripneustes gratilla (Toxopneustidae, Echinoids) is a source of protein for many islanders in the Indo-West Pacific. It was previously reported to occasionally cause ciguatera-like poisoning; however, the exact nature of the causative agent was not confirmed. In April and July 2015, ciguatera poisonings were reported following the consumption of T. gratilla in Anaho Bay (Nuku Hiva Island, Marquesas archipelago, French Polynesia). Patient symptomatology was recorded and sea urchin samples were collected from Anaho Bay in July 2015 and November 2016. Toxicity analysis using the neuroblastoma cell–based assay (CBA-N2a) detected the presence of ciguatoxins (CTXs) in T. gratilla samples. Gambierdiscus species were predominant in the benthic assemblages of Anaho Bay, and G. polynesiensis was highly prevalent in in vitro cultures according to qPCR results. Liquid chromatography–tandem mass spectrometry (LC-MS/MS) analyses revealed that P-CTX-3B was the major ciguatoxin congener in toxic sea urchin samples, followed by 51-OH-P-CTX-3C, P-CTX-3C, P-CTX-4A, and P-CTX-4B. Between July 2015 and November 2016, the toxin content in T. gratilla decreased, but was consistently above the safety limit allowed for human consumption. This study provides evidence of CTX bioaccumulation in T. gratilla as a cause of ciguatera-like poisoning associated with a documented symptomatology.

Immune effects of the neurotoxins ciguatoxins and brevetoxins

Ciguatoxins (CTXs) and brevetoxins (PbTxs) are phycotoxins that can accumulate along the marine food chain and thus cause seafood poisoning in humans, namely “ciguatera fish poisoning” (CFP) and “neurotoxic shellfish poisoning” (NSP), respectively. CFP is characterized by early gastrointestinal symptoms and typical sensory disorders (paraesthesia, pain, pruritus and cold dysaesthesia), which can persist several weeks and, in some cases, several months or years. NSP is considered a mild form of CFP with similar but less severe symptoms. After inhaled exposure, PbTxs can also cause respiratory tract irritation in healthy subjects and asthma exacerbations in predisposed subjects, whose respiratory functions may be disrupted for several days following PbTx inhalation. Mechanistically, it is well established that CTX- or PbTx-induced disturbances are primarily mainly due to voltage-gated sodium channel activation in sensory and motor peripheral nervous system. However, little is known about the pathophysiology or a potential individual susceptibility to long lasting effects of CFP/NSP. In addition to their action on the nervous system, PbTxs and CTXs were also shown to exert effects on the immune system. However, their role in the pathophysiology of syndromes induced by CTX or PbTx exposure is poorly documented. The aim of this review is to inventory the literature thus far on the inflammatory and immune effects of PbTxs and CTXs.

Pacific Ciguatoxin Induces Excitotoxicity and Neurodegeneration in the Motor Cortex Via Caspase 3 Activation: Implication for Irreversible Motor Deficit.

Consumption of fish containing ciguatera toxins or ciguatoxins (CTXs) causes ciguatera fish poisoning (CFP). In some patients, CFP recurrence occurs even years after exposure related to CTXs accumulation. Pacific CTX-1 (P-CTX-1) is one of the most potent natural substances known that causes predominantly neurological symptoms in patients; however, the underlying pathogenies of CFP remain unknown. Using clinically relevant neurobehavioral tests and electromyography (EMG) to assess effects of P-CTX-1 during the 4months after exposure, recurrent motor strength deficit occurred in mice exposed to P-CTX-1. We detected irreversible motor strength deficits accompanied by reduced EMG activity, demyelination, and slowing of motor nerve conduction, whereas control unexposed mice fully recovered in 1month after peripheral nerve injury. Finally, to uncover the mechanism underlying CFP, we detected reduction of spontaneous firing rate of motor cortical neurons even 6months after exposure and increased number of glial fibrillary acidic protein (GFAP)-immunoreactive astrocytes. Increased numbers of motor cortical neuron apoptosis were detected by dUTP-digoxigenin nick end labeling assay along with activation of caspase 3. Taken together, our study demonstrates that persistence of P-CTX-1 in the nervous system induces irreversible motor deficit that correlates well with excitotoxicity and neurodegeneration detected in the motor cortical neurons.

Ciguatoxins activate the Calcineurin signalling pathway in Yeasts: Potential for development of an alternative detection tool?

Ciguatoxins (CTXs) are lipid-soluble polyether compounds produced by dinoflagellates from the genus Gambierdiscus spp. typically found in tropical and subtropical zones. This endemic area is however rapidly expanding due to environmental perturbations, and both toxic Gambierdiscus spp. and ciguatoxic fishes have been recently identified in the North Atlantic Ocean (Madeira and Canary islands) and Mediterranean Sea. Ciguatoxins bind to Voltage Gated Sodium Channels on the membranes of sensory neurons, causing Ciguatera Fish Poisoning (CFP) in humans, a disease characterized by a complex array of gastrointestinal, neurological, neuropsychological, and cardiovascular symptoms. Although CFP is the most frequently reported non bacterial food-borne poisoning worldwide, there is still no simple and quick way of detecting CTXs in contaminated samples. In the prospect to engineer rapid and easy-to-use CTXs live cells-based tests, we have studied the effects of CTXs on the yeast Saccharomyces cerevisiae, a unicellular model which displays a remarkable conservation of cellular signalling pathways with higher eukaryotes. Taking advantage of this high level of conservation, yeast strains have been genetically modified to encode specific transcriptional reporters responding to CTXs exposure. These yeast strains were further exposed to different concentrations of either purified CTX or micro-algal extracts containing CTXs. Our data establish that CTXs are not cytotoxic to yeast cells even at concentrations as high as 1μM, and cause an increase in the level of free intracellular calcium in yeast cells. Concomitantly, a dose-dependent activation of the calcineurin signalling pathway is observed, as assessed by measuring the activity of specific transcriptional reporters in the engineered yeast strains. These findings offer promising prospects regarding the potential development of a yeast cells-based test that could supplement or, in some instances, replace current methods for the routine detection of CTXs in seafood products.

Toxicology of Gambierdiscus spp. (Dinophyceae) from Tropical and Temperate Australian Waters.

Ciguatera Fish Poisoning (CFP) is a human illness caused by the consumption of marine fish contaminated with ciguatoxins (CTX) and possibly maitotoxins (MTX), produced by species from the benthic dinoflagellate genus Gambierdiscus. Here, we describe the identity and toxicology of Gambierdiscus spp. isolated from the tropical and temperate waters of eastern Australia. Based on newly cultured strains, we found that four Gambierdiscus species were present at the tropical location, including G. carpenteri, G. lapillus and two others which were not genetically identical to other currently described species within the genus, and may represent new species. Only G. carpenteri was identified from the temperate location. Using LC-MS/MS analysis we did not find any characterized microalgal CTXs (P-CTX-3B, P-CTX-3C, P-CTX-4A and P-CTX-4B) or MTX-1; however, putative maitotoxin-3 (MTX-3) was detected in all species except for the temperate population of G. carpenteri. Using the Ca2+ influx SH-SY5Y cell Fluorescent Imaging Plate Reader (FLIPR) bioassay we found CTX-like activity in extracts of the unidentified Gambierdiscus strains and trace level activity in strains of G. lapillus. While no detectable CTX-like activity was observed in tropical or temperate strains of G. carpenteri, all species showed strong maitotoxin-like activity. This study, which represents the most comprehensive analyses of the toxicology of Gambierdiscus strains isolated from Australia to date, suggests that CFP in this region may be caused by currently undescribed ciguatoxins and maitotoxins.

First steps to develop the Surveillance Net in Macaronesian Detection of Ciguatera in species of fishing interest.

First steps to develop the Surveillance Net in Macaronesian Detection of Ciguatera in species of fishing interest.

Incidence of Ciguatoxin fish poisoning in Trivandrum, India

Ciguatoxin (CTX) is a visibly unidentifiable, colourless, odourless, heat stable and lipid soluble polyether marine biotoxin associated with human illness. Marine dinoflagellates under the genus Gambierdiscus are responsible for producingciguatoxins (CTX). The ciguatoxin gets accumulated in herbivorous fishes, gets biotransformed in carnivorous fishes and finally reach fish consumers. In January 2016, individuals who consumed red snapper in Trivandrum, Kerala, India weresuspected to be intoxicated with ciguatera based on characteristic symptoms as assessed by medical team from Trivandrum Medical College, Kerala, India. The red snapper species was identified and confirmed as Lutjanus bohar by DNA barcoding. Mouse bioassay was carried out to detect the presence of ciguatoxin and the tested mice showed symptoms related to suspected CTX toxicity. Significantly higher level of ciguatoxin lethal dose was estimated which was found equivalent to 16.25 ng of CTX-1 which led to 13% of weight loss in tested mice. Medical professionals also investigated clinical manifestations of suspected toxicity in hospitalised individuals. This study indicated that there is a need for regular surveillance of seafood landed across the coast and consumer’s awareness for their safety.

Tectus niloticus (Tegulidae, Gastropod) as a Novel Vector of Ciguatera Poisoning: Detection of Pacific Ciguatoxins in Toxic Samples from Nuku Hiva Island (French Polynesia).

Ciguatera fish poisoning (CFP) is a foodborne disease caused by the consumption of seafood (fish and marine invertebrates) contaminated with ciguatoxins (CTXs) produced by dinoflagellates in the genus Gambierdiscus. The report of a CFP-like mass-poisoning outbreak following the consumption of Tectus niloticus (Tegulidae, Gastropod) from Anaho Bay on Nuku Hiva Island (Marquesas archipelago, French Polynesia) prompted field investigations to assess the presence of CTXs in T. niloticus. Samples were collected from Anaho Bay, 1, 6 and 28 months after this poisoning outbreak, as well as in Taiohae and Taipivai bays. Toxicity analysis using the neuroblastoma cell-based assay (CBA-N2a) detected the presence of CTXs only in Anaho Bay T. niloticus samples. This is consistent with qPCR results on window screen samples indicating the presence of Gambierdiscus communities dominated by the species G. polynesiensis in Anaho Bay. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses revealed that P-CTX-3B was the major congener, followed by P-CTX-3C, P-CTX-4A and P-CTX-4B in toxic samples. Between July 2014 and November 2016, toxin content in T. niloticus progressively decreased, but was consistently above the safety limit recommended for human consumption. This study confirms for the first time T. niloticus as a novel vector of CFP in French Polynesia.

Application of solid phase adsorption toxin tracking (SPATT) devices for the field detection of Gambierdiscus toxins

Ciguatera fish poisoning is a food-borne illness caused by the consumption of seafood contaminated with ciguatoxins (CTXs) produced by dinoflagellates in the Gambierdiscus genus. Since most surveillance programs currently rely on the survey of Gambierdiscus cell densities and species composition, supplementary toxin-based methods allowing the time- and spatially integrated sampling of toxins in ciguateric environments are needed for a more reliable assessment and management of the risks associated with Gambierdiscus proliferation. Solid Phase Adsorption Toxin Tracking (SPATT) filters use porous synthetic resins capable of adsorbing toxins directly from the water column. To assess the ability of these passive monitoring devices to retain Gambierdiscus toxins, SPATT bags filled with 10g of HP20 resin were deployed for 48h in two French Polynesian locations at high (Nuku Hiva Island) vs. low to moderate (Kaukura Atoll) risk of ciguatera. CTXs could be detected in SPATT bags extracts from Nuku Hiva Island, as assessed by the mouse neuroblastoma cell-based assay (CBA-N2a) and liquid chromatography – tandem mass spectrometry (LC–MS/MS) analyses. Results of in vitro experiments suggest that the saturation limit of CTXs on HP20 resin, for a deployment time of 48h, is ≃ 55ng P-CTX-3C equiv. g−1 resin. Despite the non detection of maitotoxin (MTX), LC–MS/MS analyses showed that two other compounds also produced by Gambierdiscus species were retained on SPATT bags, i.e. iso-P-CTX-3B/C and a putative MTX analogue, known as MTX-3. This study, the first to demonstrate the suitability of SPATT technology for the in situ monitoring of Gambierdiscus toxins, highlights the potential application of this tool for routine ciguatera risk assessment and management programs.