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An aquarium hobbyist poisoning: Identification of new palytoxins in Palythoa cf. toxica and complete detoxification of the aquarium water by activated carbon
Abstract
Palytoxin (PLTX) is a lethal natural toxin often found in Palythoa zoantharians that, together with its congeners, may induce adverse effects in humans after inhalation of toxic aerosols both in open-air and domestic environments, namely in the vicinity of public and private aquaria. In this study, we describe a poisoning of an aquarium hobbyist who was hospitalized after handling a PLTXs-containing zoantharian hexacoral. Furthermore, we provide evidence for water detoxification. The zoantharian was morphologically and genetically identified as Palythoa cf. toxica (Cnidaria: Anthozoa). Palytoxin itself and two new PLTX congeners, a hydroxyPLTX and a deoxyPLTX, were detected and structurally identified by liquid chromatography high resolution multiple stage mass spectrometry (LC-HRMS(n), n = 1, 2). Total and individual toxins were quantified by LC-HRMS and sandwich ELISA both in the zoantharian (93.4 and 96.80 μg/g, respectively) and in the transport water (48.3 and 42.56 μg/mL, respectively), with an excellent mean bias of 1.3% between the techniques. Activated carbon adsorbed 99.7% of PLTXs contained in the seawater and this represents a good strategy for preventing aquarium hobbyist poisonings.
Supplementary resource (1)
... 3,8,22,23 To our knowledge, there are fourteen published reports of inhalational PTX exposure comprising forty-four cases. 3,9,22,[24][25][26][27][28][29][30][31][32][33] This represents the most reported exposure route in the literature. Of these, nearly all are associated with inhalation of steam or vapor after attempting to clean or remove zoanthid or palythoa from home aquariums with the use of hot or boiling water. ...
... The cases occur worldwide, but the majority of reported cases have occurred in the United States. The presenting symptoms are most commonly dyspnea, 3,9,22,24,25,27,28,[30][31][32] chest pain, 3,22,27 cough, 24,25,27,28,30,32,34 tachycardia, 3,9,24,27,28,30,33 nausea and/or vomiting, 3,27,29,30,33 headaches, 3,30 fever, 9,24,25,[28][29][30][31][32][33] and myalgia or weakness. 9,25,26,28,29,31,32 Since there is no antitoxin for PTX all patients received symptomatic treatment and all recovered within one month with three exceptions (one patient recovered in three months, one had recurring symptoms after two years, and one had permanent neurologic damage and respiratory symptoms after seven years of follow-up). ...
... The cases occur worldwide, but the majority of reported cases have occurred in the United States. The presenting symptoms are most commonly dyspnea, 3,9,22,24,25,27,28,[30][31][32] chest pain, 3,22,27 cough, 24,25,27,28,30,32,34 tachycardia, 3,9,24,27,28,30,33 nausea and/or vomiting, 3,27,29,30,33 headaches, 3,30 fever, 9,24,25,[28][29][30][31][32][33] and myalgia or weakness. 9,25,26,28,29,31,32 Since there is no antitoxin for PTX all patients received symptomatic treatment and all recovered within one month with three exceptions (one patient recovered in three months, one had recurring symptoms after two years, and one had permanent neurologic damage and respiratory symptoms after seven years of follow-up). ...
Palytoxin is one of the most lethal natural toxins ever discovered. This molecule has been isolated from various marine animals, including zoanthid corals. This popular organism is commonly found in many home saltwater aquariums due to its beauty and survivability. As a result of an increase in popularity, an increased number of individuals are at risk for exposure to this potentially deadly toxin. Affected patients may experience various symptoms based on the route of exposure (ie, cutaneous contact, inhalation of aerosolized toxin, ocular exposure, or ingestion). Ocular exposure can occur in various ways (eg, contact with contaminated water, rubbing the eye with a dirtied hand, or direct spraying into the eye), and incidence rates have dramatically risen in recent years. In this review, we discuss a case of systemic toxicity from inhalation and ocular exposure to presumed palytoxin on a zoanthid coral which resulted in an intensive care unit (ICU) stay, and corneal perforation which required a corneal transplant. Additionally, we review what is known about the mechanism of action of this toxin, propose a comprehensive hypothesis of its effects on corneal cells, and discuss the prognosis and clinical management of patients with systemic symptoms secondary to other routes of exposure.
... The threat of PTX poisoning is evident from numerous cases reported around the world (3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14). Its toxicity depends on the route of entry into the body. ...
... Contact an aquarium specialist and/or special services dealing with the removal of hazardous substances in your area for further instructions. Some sources claim that one of the most effective ways to remove almost all PTX from water is to use activated carbon (14). If you are working with an aquarium or its equipment you should always stay in a well-ventilated area and, if possible, open the windows to cause a draught. ...
... Schulz M, et al. Inhalation poisoning with palytoxin from aquarium coral: case description and safety advice Arh Hig Rada Toksikol 2019;70:[14][15][16][17] ...
Palythoa spp. corals and some other marine organisms contain one of the most poisonous substances ever known-palytoxin (PTX). Due to their modest life requirements and ease of breeding, these corals are popular in home aquariums. Here we refer to a case of PTX poisoning of a middle-aged woman who inhaled poisonous vapours while brushing the corals from live rock and compare it with the available literature. As the case revealed that the symptoms of PTX poisoning are not specific and neither is treatment, our aim was to give a brief tabulated review of the symptoms that may indicate such poisoning. Cases of palytoxin poisoning have been reported worldwide, and severe ones (mostly due to ingestion of contaminated sea food) can end in death. As it appears, most (if not all) poisonings result from unawareness of the risk and reckless handling by aquarists. This is one of the first articles which provides some practical advice about the use of personal protection equipment, including gloves, masks, eyewear, and other clothing during any coral manipulation to minimise the risk. We also draw attention to the lack of marketing/trading regulations for dangerous coral species and/ or regulations or instructions dealing with their removal and health protection.
... In addition to palytoxin, a series of its congeners has been identified so far. They differ from palytoxin for additional and/or missing hydroxyl and/or methyl groups, or for chiralities, which sometimes influence their toxic potency (Tartaglione et al. 2016). Only few palytoxin analogs have been studied under a chemical and/or biological point of view. ...
... Based on all animal studies, Riobó et al. (2006) reported the estimated dose of palytoxin for humans to be between 2.3 and 31.5 μg, and an ARfD was suggested to be 64 μg for a person with an average weight of 60 kg (Rhodes et al. 2008). Recent evidence also suggests that humans are poisoned by palytoxin and its congeners through inhalation and dermal routes (Majlesi et al. 2008;Nordt et al. 2009;Hall et al. 2015;Tartaglione et al. 2016). The most common signs after inhalation and cutaneous/systemic exposures are respiratory distress, bronchoconstriction, mild dyspnea, rhinorrhea, cough, fever, and a small incidence of dermatitis and conjunctivitis (Thakur and Jha 2017). ...
Palytoxin, isolated from a zoanthid of the genus Palythoa, is the most potent marine toxin known. Intoxication by palytoxin leads to vasoconstriction, hemorrhage, ataxia, muscle weakness, ventricular fibrillation, pulmonary hypertension, ischemia and death. Palytoxin and its numerous derivatives (congeners) may enter the food chain and accumulate mainly in fishes and crabs, causing severe human intoxication and death following ingestion of contaminated products. Furthermore, toxic effects in individuals exposed via inhalation or skin contact to marine aerosol in coincidence with Ostreopsis blooms, have been reported. Blooms of the benthic dinoflagellate Ostreopsis cf. ovata are a concern in the Mediterranean Sea, since this species produces a wide range of palytoxin-like compounds listed among the most potent marine toxins. Thus, the formerly unsuspected broad distribution of the benthic dinoflagellate Ostreopsis spp. has recently posed a problem of risk assessment for human health. Palytoxin has a strong potential for toxicity in humans and animals, and currently this toxin is of great concern worldwide. This review summarized and discussed the pharmacology and toxicology data of palytoxin and its congeners, including their cytotoxicity, human and animal toxicities. Moreover, the risk assessment and their control strategies including prevention and treatment assays were evaluated.
... Des antécédents respiratoires et/ou cardiovasculaires pourraient être un facteur de risque de gravité lors d'une intoxication à la palytoxine. La symptomatologie présentée par les patients de cette étude en fonction des voies d'exposition correspond à ce qui est retrouvé dans la littérature pour les voies respiratoire [2,11,15,[19][20][21][22][23][24], cutanée [1,8,14,25], et oculaire [5,[26][27][28]. Enfin, comme vu plus haut, que ce soit dans notre série de cas ou la littérature, les manipulateurs de coraux mous des aquariums d'eau de mer ne portent pas d'EPI, ou bien seulement de façon partielle, tout comme les personnes à proximité de la manipulation. ...
... • masque complet équipé de filtre(s) anti-aérosols P3 (norme EN149, R ou NR) [19,22,29,30] ; • gants enduits avec manchettes, résistants contre les risques chimiques (norme EN374) et de coupure (norme EN388) [31] ; • combinaison de protection de type 4 avec capuche. ...
Background: Following a French cases report publication (2016) concerning palytoxin poisoning during cnidarians manipulation (Palythoa, Zoanthus), the French Agency for Food, Environmental and Occupational Health & Safety (ANSES) took interest. Objective: Identification and description of palytoxin exposure cases, during cnidarian manipulation in an aquarium, brought to the french anti-poisoning centers in purpose to advise sanitary authorities and suggest preventive measures. Method: Retrospective descriptive study of palytoxin exposure cases brought to the French anti-poisoning centers between the 2000/01/01 and the 2017/12/31, following the objective. Results: In all, 23 cases, in 14 files, were included. Exposure concerned 16 private individuals and 7 professional individuals. A multi-organ exposure was predominant: respiratory; cutaneous and/or ocular. All the cases were symptomatic, with multi-organs affection: no specific medical signs (17 cases), neurologic signs (14 cases), digestive signs (11 cases), respiratory signs (10 cases), ocular signs (8 cases) and/or cutaneous signs (7 cases). Severity of illness was minor for 14 cases, moderate for 8 cases and major for 1 case (corneal ulcer with sequela). Only four professional individuals have worn personal protective equipment, partially. Conclusion: Palytoxin exposure affects private and professional individuals of fishkeeping sector, with multi-organs exposure. Severity is minor in most of the cases, but can be major with sequela. It is significant to advise individuals about risks during cnidarians manipulation and protective measures that exist.
... Dès lors, la PLTX et les PLTX-like ont été retrouvées dans divers coraux mous des genres Palythoa et Zoanthus. Leur présence a également été prouvée dans des dinoflagellés du genre Ostreopsis, dans certaines cyanobactéries et au sein de poissons, mollusques, échinodermes et crustacés susceptibles d'accumuler dans leurs tissus les PLTX-like [1][2][3]. La PLTX a une structure non protéique, d'environ 2600 Da [1]. Elle est thermostable et est 10 à 100 fois plus toxique que la tétrodotoxine, toxine responsable du FUGU [4]. ...
... Le risque associé à la manipulation des coraux d'aquarium est connu en Amérique du Nord [10], mais il ne semble pas y avoir à ce jour aux États-Unis de contrôle particulier ou de message de prévention associé à leur vente. En Europe, les aquariophiles et les revendeurs d'équipement d'aquarium ne semblent pas être sensibilisés à cette problématique et à notre connaissance il n'y a aucune régulation particulière lors de l'importation ou la revente de ces cnidaires ornementaux [1,3]. ...
... margaritae, P. canariensis, P. heliodiscus and P. aff. clavata/sakurajimensis. 2,6,17,[31][32][33][34][35][36][37][38][39] The concentration of palytoxin (32) in P. aff. clavata/sakurajimensis is one of the highest reported in the literature, with 2.22 mg g À1 of wet sample, followed by P. heliodiscus reported by Deeds et al., with a concentration of 1 mg g À1 wet sample, while deoxypalytoxin (36) was found at 3.51 mg g À1 wet sample. ...
Covering: up to the end of 2018
Zoantharians, also improperly known as zoanthids or colonial anemones, are well known by aquarists because of their ease of use in aquaria but also because of their splendid colours. However, high concentrations of the highly toxic palytoxin found in some species of zoantharians maintained in reef aquaria has raised some issues recently, unveiling at the same time a rather unknown chemical diversity hidden in these marine beauties. Herein, we report the structure of the metabolites described in all species of zoantharians up to the end of 2018 and their associated biological activities. As sessile invertebrates, zoantharians harbour a rich diversity of micro-organisms that can play a role in the biosynthesis of these natural products and we detail the current hypotheses on the metabolic pathways leading to the identified ecdysteroids, zoanthoxanthins, zoanthamines, palytoxins and others. Finally, we assess the possible use of these metabolites in the systematics of such a complex group of marine invertebrates and we discuss their possible ecological roles. Altogether, this review brings some insights into the rich chemical diversity of zoantharians and their potential for marine biodiscovery and marine ecology.
... Ocular exposure to palytoxin is rare. Aquarium water filtered through activated charcoal reduces palytoxin levels by 99.4% [12], and ocular exposure occurs after handling coral above water, by auto-contamination or direct ocular contact with water jets siphoned by coral [13][14][15][16]. ...
... This lack of information poses serious threats to aquarists because it is unknown what species are currently being distributed through the aquarium trade, their potential toxicity, or their human health risks. There have been multiple reports of accidental poisoning through the marine aquarium trade, however inconsistent identification and taxonomic uncertainty of Palythoa species impede our understanding of the risks to aquarium hobbyists (Hoffmann et al. 2008;Deeds and Schwartz 2010;Deeds et al. 2011;Tartaglione et al. 2016). Due to the current uncertain state of Palythoa taxonomy, it is unclear which species are being exported and imported around the world, along with the distribution of PTX among these species. ...
Zoantharians (Cnidaria: Hexacorallia: Zoantharia) of the genus Palythoa are ubiquitous species that occupy reef habitats in every tropical ocean. Disagreements among classifications based on morphology, reproductive traits, and molecular techniques have generated taxonomic challenges within this group. Molecular studies provide limited phylogenetic resolution between species, and discordance is frequently attributed to slow mitochondrial rates and lack of resolution among molecular markers. Here we conducted the first phylogenomic survey of Palythoa, using a reduced representation genomic approach (ezRAD) to resolve relationships among eight described and four putative Palythoa species (N = 22 plus two outgroups) across the Pacific and Atlantic Oceans. We constructed nearly complete mitochondrial genomes and assembled transcriptome loci datasets by reference mapping. A de novo assembly was performed for the holobiont dataset, and we compared a range of filtering strategies from unfiltered data down to 136 unlinked high-quality biallelic SNPs shared by all samples to resolve evolutionary lineages within Palythoa. Across all these datasets, the resulting Bayesian and ML trees revealed six highly concordant and well-supported clades, however, the phylogenomic data were inconclusive in resolving species relationships within the clades. We detected putative species complexes within two well sampled Palythoa clades (clades I and II), but species delimitation results were inconsistent in whether these clades contain multiple nominal species or represent a single variable species. Polyphyly in the broadly distributed species Palythoa tuberculosa and P. mutuki highlight the need for additional study. Consistency among nuclear and mitogenomic datasets points to a lack of biological understanding of species boundaries among these zoantharians rather than limitations of the molecular markers. More complete taxonomic sampling of nominal species across the geographic ranges of distribution is necessary to resolve species boundaries and evolutionary histories among members of this genus.
... The bright and fluorescence colors displayed by these sessile invertebrates attracts anyone's attention for decoration of aquariums. However, most of the species being sold are not assess for their toxin content and several cases of palytoxin intoxication through steam inhalation, ocular exposure or skin contact while cleaning aquariums have already been recorded [117]. Deeds et al., reported high concentrations of the toxin in four different zoantharian species sold in aquarium stores clearly demonstrating the high health-related risk of having these organisms at home [119]. ...
This thesis describes the isolation and identification of marine natural products from five zoantharian species collected off the Marine Protected Area El Pelado located in Santa Elena, Ecuador. A new family of 2-aminoimidazole alkaloids named terrazoanthines A-C were identified from Terrazoanthus patagonichus. Terrazoanthines A and B are characterized by the presence of the 2-aminoimidazole ring fused to a cyclohexane. Additionally, two zoanthoxanthin derivatives named zoamides E and F were isolated from the sister species Terrazoanthus cf. patagonichus. Then, four ecdysteroids derivatives named ecdysonelactones were obtained from Antipathozoanthus hickmani. These compounds feature a γ-lactone ring fused to ring A of the ecdysteroid skeleton. Further investigations on this organism led to the identification of four halogenated tyrosine derivatives named valdiviamides A-D and characterized by the presence of iodine and bromine atoms in the phenol ring. The bioactivity study revealed valdiviamide B to have moderate activity against the liver cancer cell line (HepG2) with an IC50 value of 7.8 µM. Additionally, two halogenated tyramine derivatives containing iodine and bromine atoms were identified from Parazoanthus darwini. The chemical investigation of Zoanthus cf. pulchellus allowed the identification of two members of the bioactive family of zoanthamine alkaloids. These compounds revealed neuroinflammatory activity in microglia BV-2 cells with high inhibitory effects in reactive oxygen species (ROS) and nitric oxide (NO) generation.
... En conséquence, la palytoxine provoque un large spectre d'actions pharmacologiques secondaires telles que la dépolarisation des membranes excitables et l'activation secondaire des canaux Ca 2+ (Ibrahim and Shier, 1987;Wu, 2009 (Biré et al., 2013;Lenoir et al., 2004;Onuma et al., 1999) reconnus comme principaux producteurs. Kerbrat et al.(2011) (Farooq et al., 2017;Pelin et al., 2016;Schmitt et al., 2018;Tartaglione et al., 2016a;Tartaglione et al., 2016b). ...
Face à l’expansion géographique des biotoxines marines, à l’émergence de nouvelles toxines et compte tenu du risque avéré pour la santé humaine, il est essentiel de disposer d’outils suffisamment versatiles et performants pour détecter une gamme, la plus large possible, de toxines connues ou émergentes de manière à garantir la sécurité des consommateurs. Cette thèse s’inscrit dans la démarche de surveillance de la qualité sanitaire des produits de la pêche. Elle a pour finalité de contribuer à l’évolution du dispositif de veille d’émergence par le développement d’une approche non ciblée reposant sur l’utilisation de la spectrométrie de masse haute résolution comme alternative à la pratique controversée du bio-essai sur souris.Les travaux entrepris ont permis dans un premier temps de développer et caractériser une méthode par chromatographie liquide couplée à la spectrométrie de masse haute résolution pour l'analyse ciblée de 32 toxines marines avec une gamme étendue de polarités, utilisant un spectromètre de masse haute résolution. Deux types de séparations chromatographiques, en phase inverse et à interactions hydrophiles, ont été mises en place pour la séparation des toxines lipophiles et hydrophiles. Ensuite une stratégie décrivant les différentes étapes d’une approche non ciblée allant de l’acquisition au traitement des données par des outils chimiométriques a été développée. Le traitement des données acquises en mode non ciblé a été réalisé au moyen de deux types de logiciels différents : une suite logicielle commerciale (Sciex) et un logiciel open source (XCMS). Cette stratégie a été testée avec succès dans le cadre d’une preuve de concept sur des échantillons d’huîtres et de moules supplémentés avec certaines toxines et analysés en aveugle. Elle a ensuite été appliquée sur des échantillons impliqués dans des cas de toxi-infections alimentaires collectives liés à la consommation de violets du genre Microcosmus, selon les deux approches différentes, le suspect screening et l’analyse sans a priori.
... The increment in toxic blooms of PLTX in Europe raised high concern and the urgent need to obtain toxicity data useful for the future regulation of this toxin in marine products captured in European coasts. Besides causing human intoxications, after consumption of marine crustaceans and fish [14][15][16][17], toxic effects of different severity have been attributed to PLTX after human inhalation of marine aerosols or skin contact during Ostreopsis blooms [18][19][20] and even after dermal or inhalatory exposition to zoanthid corals in aquariums [21,22]. ...
Palytoxin (PLTX) is one of the most poisonous substances known to date and considered as an emergent toxin in Europe. Palytoxin binds to the Na+-K+ ATPase, converting the enzyme in a permeant cation channel. This toxin is known for causing human fatal intoxications associated with the consumption of contaminated fish and crustaceans such as crabs, groupers, mackerel, and parrotfish. Human intoxications by PLTX after consumption of contaminated fishery products are a serious health issue and can be fatal. Different reports have previously explored the acute oral toxicity of PLTX in mice. Although the presence of palytoxin in marine products is currently not regulated in Europe, the European Food Safety Authority expressed its opinion on PLTX and demanded assessment for chronic toxicity studies of this potent marine toxin. In this study, the chronic toxicity of palytoxin was evaluated after oral administration to mice by gavage during a 28-day period. After chronic exposure of mice to the toxin, a lethal dose 50 (LD50) of 0.44 µg/kg of PLTX and a No-Observed-Adverse-Effect Level (NOAEL) of 0.03 µg/kg for repeated daily oral administration of PLTX were determined. These results indicate a much higher chronic toxicity of PLTX and a lower NOAEL than that previously described in shorter treatment periods, pointing out the need to further reevaluate the levels of this compound in marine products.
... During the last decades, benthic dinoflagellates of the genus Ostreopsis, usually detected in tropical areas, have expanded their distribution to temperate waters and were recently identified in some European countries such as France, Greece, Italy, Cyprus, Spain and Portugal (Accoroni et al., 2016;Aligizaki et al., 2008;Ciminiello et al., 2006;Del Favero et al., 2013;Fraga et al., 2017;Santos et al., 2019) causing human intoxications associated with the consumption of contaminated fish and crustaceans as well as to exposure to marine aerosols (Patocka et al., 2018;Pelin et al., 2016;Walsh et al., 2017) and even contact with corals in aquariums (Hoffmann et al., 2008;Tartaglione et al., 2016). ...
Palytoxin is an emergent toxin in Europe and one of the most toxic substances know to date. The toxin disrupts the physiological functioning of the Na+/K+-ATPase converting the enzyme in a permeant cation channel. Human intoxications by PLTX after consumption of contaminated fishery products are a serious health issue and can be fatal. Several reports have previously investigated the oral and intraperitoneal toxicity of PLTX in mice. However, in all cases short observation periods (24 and 48 h) after toxin administration were evaluated. In this work, single oral or intraperitoneal doses of PLTX were administered to healthy mice and surviving animals were followed up for 96 h. The data obtained here allowed us to calculate the oral and intraperitoneal lethal doses 50 (LD50) which were in the range of the values previously described. Surprisingly, the oral NOAEL for PLTX was more than 10 times lower than that previously described, a fact that indicates the need for the reevaluation of the levels of the toxin in edible fishery products.
... This lack of information poses serious threats to aquarists because it is unknown what species are currently being distributed through the aquarium trade, their potential toxicity, or their human health risks. There have been multiple reports of accidental poisoning through the marine aquarium trade, however inconsistent identification and taxonomic uncertainty of Palythoa species impede our understanding of the risks to aquarium hobbyists (Hoffmann et al. 2008;Deeds and Schwartz 2010;Deeds et al. 2011;Tartaglione et al. 2016). Due to the current uncertain state of Palythoa taxonomy, it is unclear which species are being exported and imported around the world, along with the distribution of PTX among these species. ...
Zoantharians (Cnidaria: Hexacorallia: Zoan-tharia) of the genus Palythoa are ubiquitous species that occupy reef habitats in every tropical ocean. Disagreements among classifications based on morphology, reproductive traits, and molecular techniques have generated taxonomic challenges within this group. Molecular studies provide limited phylogenetic resolution between species, and dis-cordance is frequently attributed to slow mitochondrial rates and lack of resolution among molecular markers. Here we conducted the first phylogenomic survey of Pa-lythoa, using a reduced representation genomic approach (ezRAD) to resolve relationships among eight described and four putative Palythoa species (N = 22 plus two out-groups) across the Pacific and Atlantic Oceans. We constructed nearly complete mitochondrial genomes and assembled transcriptome loci datasets by reference mapping. A de novo assembly was performed for the holobiont dataset, and we compared a range of filtering strategies from unfiltered data down to 136 unlinked high-quality biallelic SNPs shared by all samples to resolve evolutionary lineages within Palythoa. Across all these datasets, the resulting Bayesian and ML trees revealed six highly con-cordant and well-supported clades, however, the phyloge-nomic data were inconclusive in resolving species relationships within the clades. We detected putative species complexes within two well sampled Palythoa clades (clades I and II), but species delimitation results were inconsistent in whether these clades contain multiple nominal species or represent a single variable species. Polyphyly in the broadly distributed species Palythoa tuberculosa and P. mutuki highlight the need for additional study. Consistency among nuclear and mitogenomic data-sets points to a lack of biological understanding of species boundaries among these zoantharians rather than limitations of the molecular markers. More complete taxonomic sampling of nominal species across the geographic ranges of distribution is necessary to resolve species boundaries and evolutionary histories among members of this genus.
Marine biotoxins, mainly produced by microalgae, show a wide range of structural diversity from simple amino acids to some of the largest and most complex natural products identified to date. Moreover, they display remarkable biological activities, and some compounds are even considered the most potent nonproteogenic venoms. This chapter presents the structures, analogues and structure-activity relationships of most marine natural products biosynthesized by microalgae with toxic properties towards humans and/or marine organisms. The chapter includes excitatory amino acids (domoic acid and derivates, kainic acid and β-N-methylamino-alanine), alkaloids (saxitoxins and tetrodotoxins) and four main groups of polyketides: linear and macrocycled polyethers (okadaic acid and derivates, pectenotoxins, azaspiracids and goniodomins), polyether ladders (yessotoxins, brevetoxins, brevisulcenals, brevisulcatic acids, gymnocins, ciguatoxins, maitotoxins, gambieric acids, gambierol, gambieroxide, gambierone and prymesins), cyclic imines (spirolides, gymnodimines, pinnatoxins and pteriatoxins, prorocentrolides, spiro-prorocentrimine, symbioimine and portimine) and long carbon-chain polyols (palytoxins, ostreocins, mascarenotoxins, ovatoxins, ostreol, amphidinols and karlotoxins).
Anemone-like animals belonging to the order Zoantharia are common anthozoans widely distributed from shallow to deep tropical and subtropical waters. Some species are well-known because of their high toxicity due to the presence of palytoxin (PLTX) in their tissues. PLTX is a large polyhydroxylated compound and one of the most potent toxins known. Currently, the PLTX biosynthetic pathway in zoantharians and the role of the host or the putative symbiotic organism(s) involved in this pathway are entirely unknown. To better understand the presence of PLTX in some Zoantharia, twenty-nine zoantharian colonies were analysed in this study. All zoantharian samples and their endosymbiotic dinoflagellates (Symbiodiniaceae = Zooxanthellae) were identified using DNA barcoding and phylogenetic reconstructions. Quantification of PLTX and its analogues showed that the yields contained in Palythoa heliodiscus, Palythoa aff. clavata and one potentially undescribed species of Palythoa are among the highest ever found (up to > 2 mg/g of wet zoantharian). Mass spectrometry imaging was used for the first time on Palythoa samples and revealed that in situ distribution of PLTX is mainly located in ectodermal tissues such as the epidermis of the body wall and the pharynx. Moreover, high levels of PLTX have been detected in histological regions where few or no Symbiodiniaceae cells could be observed. Finally, issues such as host‐specificity and environmental variables driving biogeographical patterns of hosted Symbiodiniaceae in zoantharian lineages were discussed in light of our phylogenetic results as well as the patterns of PLTX distribution. It was concluded that (1) the variability of Symbiodiniaceae diversity may be related to ecological divergence in Zoantharia, (2) all Palythoa species hosted Cladocopium Symbiodiniaceae (formerly clade C), (3) the sole presence of Cladocopium is not sufficient to explain the presence of high concentrations of PLTX and/or its analogues, and (4) the ability to produce high levels of PLTX and/or its analogues highlighted in some Palythoa species could be a plesiomorphic character inherited from their last common ancestor and subsequently lost in several lineages.
The marine algal toxin palytoxin (PLTX) and its analogues are some of the most toxic marine compounds. Their accumulation in edible marine organisms and entrance into the food chain represent their main concerns for human health. Indeed, several fatal human poisonings attributed to these compounds have been recorded in tropical and subtropical areas. Due to the increasing occurrence of PLTX in temperate areas such as the Mediterranean Sea, the European Food Safety Authority (EFSA) has suggested a maximum limit of 30 µg PLTX/kg in shellfish meat, and has recommended the development of rapid, specific, and sensitive methods for detection and quantitation of PLTX in seafood. Thus, a novel, sensitive cell-based ELISA was developed and characterized for PLTX quantitation in mussels. The estimated limits of detection (LOD) and quantitation (LOQ) were 1.2 × 10−11 M (32.2 pg/mL) and 2.8 × 10−11 M (75.0 pg/mL), respectively, with good accuracy (bias = 2.5%) and repeatability (15% and 9% interday and intraday relative standard deviation of repeatability (RSDr), respectively). Minimal interference of 80% aqueous methanol extract allows PLTX quantitation in mussels at concentrations lower than the maximum limit suggested by EFSA, with an LOQ of 9.1 µg PLTX equivalent/kg mussel meat. Given its high sensitivity and specificity, the cell-based ELISA should be considered a suitable method for PLTX quantitation.
Zusammenfassung
Palytoxine, die in Korallen vorkommen und unter anderem durch Dinoflagellaten synthetisiert werden, wirken bei Exponierten über einen Eingriff in die Natrium-Kalium-Pumpe und führen somit zu einer Änderung des Konzentrationsgradienten zwischen intra- und extrazellulärem Raum. Die Symptomatik kann aus Atemnot, Übelkeit und Erbrechen, Kopfschmerzen, Tachykardie und EKG-Veränderungen, aber auch aus Ulzerationen der Hornhaut bestehen. Die symptomatische Therapie setzt sich zusammen aus inhalativen Bronchodilatatoren, evtl. Infusion und Schmerztherapie, Antibiotikatherapie bei Infizierung der Schnittwunden, Vorstellung beim Augenarzt und Behandlung mit Vitamin-A-haltigen sowie antibiotischen Augentropfen. Um eine Exposition mit Palytoxinen zu vermeiden, gilt es, entweder auf Korallen im Aquarium zu verzichten oder bereits vorhandene Korallen nicht aus dem Aquarium zu entfernen.
Palytoxin has been found in several soft coral species which are popular for in-home reef aquariums. Although a limited number of case reports describing acute respiratory distress after inhalational exposure to palytoxin have been reported, there have been no reports regarding the potential long-term effects of inhalational exposure to palytoxin in the literature. This case follows an aquatic specialist in the U.S. over a period of seven years after a single intense occupational exposure to the aerosolized toxin from cleaning of a residential aquarium.
Palytoxin (PLTX) is a complex marine toxin produced by Zoanthids (Palyhtoa), dinoflagellates (Ostreopsis) and cyanobacteria (Trichodesmium). Contact with PLTX-like compounds present in aerosols or marine organisms has been associated with adverse effects on humans. The worldwide distribution of producer species and seafood contaminated with PLTX-like molecules illustrates the global threat to human health. The identification of species capable of palytoxin production is critical for human safety. We studied the presence of PLTX analogues in Palythoa canariensis, a coral species collected in the Atlantic Ocean never described as a PLTX-producer before. Two methodologies were used for the detection of these toxins: a microsphere-based immunoassay that offered an estimation of the content of PLTX-like molecules in a Palythoa canariensis extract and an ultra-high pressure liquid chromatography coupled to ion trap with time of flight mass spectrometer (UPLC-IT-TOF-MS) that allowed the characterization of the toxin profile. The results demonstrated the presence of PLTX, hydroxy-PLTX and, at least, two additional compounds with PLTX-like profile in the Palythoa canariensis sample. The PLTX content was estimated in 0.27 mg/g of lyophilized coral using UPLC-IT-TOF-MS. Therefore, this work demonstrates that Palythoa canariensis produces a mixture of PLTX-like molecules. This is of special relevance to safeguard human health considering Palythoa species are commonly used for decoration by aquarium hobbyists.
Palytoxin (PLTX), one the most potent marine toxins, and/or its analogs, have been identified in different marine organisms, such as Palythoa soft corals, Ostreopsis dinoflagellates, and Trichodesmium cyanobacteria. Although the main concern for human health is PLTXs entrance in the human food chain, there is growing evidence of adverse effects associated with inhalational, cutaneous, and/or ocular exposure to aquarium soft corals contaminated by PLTXs or aquaria waters. Indeed, the number of case reports describing human poisonings after handling these cnidarians is continuously increasing. In general, the signs and symptoms involve mainly the respiratory (rhinorrhea and coughing), skeletomuscular (myalgia, weakness, spasms), cardiovascular (electrocardiogram alterations), gastrointestinal (nausea), and nervous (paresthesia, ataxia, tremors) systems or apparates. The widespread phenomenon, the entity of the signs and symptoms of poisoning and the lack of control in the trade of corals as aquaria decorative elements led to consider these poisonings an emerging sanitary problem. This review summarizes literature data on human poisonings due to, or ascribed to, PLTX-containing soft corals, focusing on the different PLTX congeners identified in these organisms and their toxic potential.
Inhalational exposure to palytoxin is an extremely rare cause of respiratory distress. This little-known marine toxin has the potential to cause significant morbidity and mortality. Toxicity has been best documented in cases of ingestion but has also been seen in cases of dermal exposure and inhalation of vapors. Palytoxin has been found in several coral species, some of which are favored by home aquarium enthusiasts and are commercially available. We report a case of a family who were exposed to the aerosolized toxin following the cleaning of a coral in their home aquarium. It is important that clinicians be aware of this source of toxic exposure to provide necessary care to these patients.
Palytoxins from Ostreopsis cf. ovata (a putative palytoxin and ovatoxins) are emerging toxins in the Mediterranean basin and are not yet regulated, although there is evidence that they can accumulate in seafood and thus enter the human food chain. This poses serious concerns for human health, because palytoxin itself is among the most potent marine toxins known. In 2009, the European Food Safety Authority (EFSA) announced the need for optimization of efficient analytical methods for detecting palytoxins and for preparing standards. Herein, we propose a procedure including a one-step extraction, solid-phase-extraction (SPE) clean-up, and liquid chromatography-high resolution mass spectrometry (LC–HRMS) detection of individual palytoxins in mussels. The method enabled efficient chromatographic separation of individual compounds, including structural isomers, with good sensitivity, reproducibility, and linearity in a large dynamic range (14–1000 ng mL−1 in matrix). As a result, the putative palytoxin from Ostreopsis cf. ovata was identified as an isomer of palytoxin itself and re-named isobaric palytoxin. The whole procedure (sample preparation and LC–HRMS analysis) proved able to detect palytoxins in both spiked and natural mussel samples at levels as low as 70 μg kg−1 in crude mussel extracts and 15 μg kg−1 after SPE clean-up. Although full validation of the method is currently prevented by the unavailability of palytoxin(s) certified standards and reference material, this study constitutes a first step towards achieving this.
Despite the Central Indo-Pacific (CIP) and the Indonesian Archipelago being a well-known region of coral reef biodiversity, particularly in the 'Coral Triangle', little published information is available on its zoantharians (Cnidaria: Hexacorallia: Zoantharia). In order to provide a basis for future research on the Indo-Pacific zoantharian fauna and facilitate comparisons between more well-studied regions such as Japan and the Great Barrier Reef, this report deals with CIP zoantharian specimens in the Naturalis collection in Leiden, the Netherlands; 106 specimens were placed into 24 morpho-species and were supplemented with 88 in situ photographic records from Indonesia, the Philippines, and Papua New Guinea. At least nine morpho-species are likely to be undescribed species, indicating that the region needs more research in order to properly understand zoantharian diversity within the CIP. The Naturalis' zoantharian specimens are listed by species, as well as all relevant collection information, and in situ images are provided to aid in future studies on zoantharians in the CIP.
Since the late 1990's, a respiratory syndrome has been repetitively observed in humans concomitant with Ostreopsis spp. blooms (mainly O. cf. ovata) in the Mediterranean area. Previous studies have demonstrated that O. cf. ovata produces analogues of palytoxin (ovatoxins and a putative palytoxin), one of the most potent marine toxins. Based on the observed association between O. cf. ovata blooms, respiratory illness in people, and detection of palytoxin complex in algal samples, toxic aerosols - containing Ostreopsis cells and/or the toxins they produce - were postulated to be the cause of human illness. A small scale monitoring study of marine aerosol carried out along the Tuscan coasts (Italy) in 2009 and 2010 is reported. Aerosols were collected concurrently O. cf. ovata blooms and they were analyzed by both PCR assays and LC-HRMS technique. The results, besides confirming the presence of O. cf. ovata cells, demonstrated for the first time the occurrence of ovatoxins in the aerosol at levels of 2.4 pg of ovatoxins per air liter. Given the lack of toxicological data on palytoxins by inhalation exposure, our results are only a first step towards a more comprehensive understanding of the Ostreopsis-related respiratory syndrome.
The genus Protopalythoa Verrill, 1900 (family Sphenopidae) is defined and its status discussed. As with other zoanthid genera, species are difficult to separate by traditional methods. We have developed a rigorous approach to quantitative data derived from measurements and meristic characters. In support of genetic evidence, we have utilised the relationship between number of septa and polyp column diameter, and the capsule length of nematocysts (holotrichs and p-mastigophores), together with the presence or absence of basitrichs in the cnidom. Using this approach, the species Pr. mutuki (Haddon & Shackleton, 1891b) and Pr. heliodiscus, sp. nov. are separated, described and illustrated. Protopalythoa mutuki is interpreted as a normal micro-carnivore with autotrophic capability, while Pr. heliodiscus sp. nov. is an obligate autotroph with little ability to capture prey. The latter is the only zoanthid known to have vertical transmission of zooxanthellae.
Introduction:
Palytoxin (PTX) is considered a severe marine toxin. Although rare, reports of human exposure from consumption of PTX have described significant morbidity and mortality. PTX is the suspected agent in Haff disease, in which rhabdomyolysis occurs within 24 h of eating contaminated fish such as buffalo fish. PTX is primarily present in soft corals or in dinoflagellates, and it can contaminate crustaceans and other fish as it bioaccumulates up the food chain. Only 23 cases have been reported in the USA, including two recent cases in New York City. Reports of inhalational exposure to PTX are uncommon.
Case reports:
We describe a case series of six patients, including four adults and two children, with inhalational exposure to PTX aerosolized from Palythoa corals. Their symptoms included some degree of respiratory involvement, myalgias, paresthesias, low-grade fevers, and gastrointestinal symptoms. Fortunately, there were no serious outcomes and all patients survived without sequelae.
Discussion:
Although rare, exposure to palytoxin is not restricted to people visiting marine environments because of Palythoa coral in some home aquariums. Routes of exposure go beyond consumption of fish that feed on the coral and include dermal as well as inhalational exposure. Palytoxin exposure should be considered in the differential diagnosis of patients who own or work with fish tanks and present with symptoms that include respiratory complaints, myalgias, neuromuscular dysfunction, hemolysis, and cardiac toxicity. There is no known antidotal therapy and treatment should focus on meticulous supportive care.
Palytoxin (PLTX) is the reference compound for a group of potent marine biotoxins, for which the molecular target is Na+/K+-ATPase. Indeed, ouabain (OUA), a potent blocker of the pump, is used to inhibit some PLTX effects in vitro. However, in an effort to explain incomplete inhibition of PLTX cytotoxicity, some studies suggest the possibility of two different binding sites on Na+/K+-ATPase. Hence, this study was performed to characterize PLTX binding to intact HaCaT keratinocytes and to investigate the ability of OUA to compete for this binding. PLTX binding to HaCaT cells was demonstrated by immunocytochemical analysis after 10 min exposure. An anti-PLTX monoclonal antibody-based ELISA showed that the binding was saturable and reversible, with a K(d) of 3 × 10-10 M. However, kinetic experiments revealed that PLTX binding dissociation was incomplete, suggesting an additional, OUA-insensitive, PLTX binding site. Competitive experiments suggested that OUA acts as a negative allosteric modulator against high PLTX concentrations (0.3-1.0 × 10-7 M) and possibly as a non-competitive antagonist against low PLTX concentrations (0.1-3.0 × 10-9 M). Antagonism was supported by PLTX cytotoxicity inhibition at OUA concentrations that displaced PLTX binding (1 × 10-5 M). However, this inhibition was incomplete, supporting the existence of both OUA-sensitive and -insensitive PLTX binding sites.
Zoantharia (or Zoanthidea) is the third largest order of Hexacorallia, characterised by two rows of tentacles, one siphonoglyph
and a colonial way of life. Current systematics of Zoantharia is based exclusively on morphology and follows the traditional
division of the group into the two suborders Brachycnemina and Macrocnemina, each comprising several poorly defined genera
and species. To resolve the phylogenetic relationships among Zoantharia, we have analysed the sequences of mitochondrial 16S
and 12S rRNA genes obtained from 24 specimens, representing two suborders and eight genera. In view of our data, Brachycnemina
appears as a monophyletic group diverging within the paraphyletic Macrocnemina. The macrocnemic genus Epizoanthus branches as the sister group to all other Zoantharia that are sampled. All examined genera are monophyletic, except Parazoanthus, which comprises several independently branching clades and individual sequences. Among Parazoanthus, some groups of species can be defined by particular insertion/deletion patterns in the DNA sequences. All these clades show
specificity to a particular type of substrate such as sponges or hydrozoans. Substrate specificity is also observed in zoantharians
living on gorgonians or anthipatharians, as in the genus Savalia (Gerardia). If confirmed by further studies, the substrate specificity could be used as reliable character for taxonomic identification
of some Macrocnemina.
A community ecology approach to the study of the most common group of zooxanthellae, dinoflagellates in the genus Symbiodinium, was applied to symbiotic invertebrate assemblages on coral reefs in the western Caribbean, off the Yucatan peninsula (Puerto Morelos, Mexico) and over 1000 km away in the northeastern Caribbean, at Lee Stocking Island, Bahamas. Sequence differences and intragenomic variation, as determined by denaturing gradient gel electrophoresis and sequencing of the internal transcribed spacer 2 (ITS 2) region, were used to classify these symbionts. Twenty-eight genetically distinct Symbiodinium types were identified, eleven of which were found in hosts from both Caribbean locations. A single symbiont population was detected in 72% of hosts from the Yucatan and 92% of hosts from the Bahamas. The reef-wide community distribution of these symbionts is dominated by a few types found in many different host taxa, while numerous rare types appear to have high specificity for a particular host species or genus. Clade or lineage A Symbiodinium spp. was restricted to compatible hosts located within 3-4 m of the surface, while Symbiodinium spp. types from other lineages displayed differences in vertical zonation correlated with ITS type but were independent of clade designation. A comparison of the symbiont types found in field-collected hosts with types previously cultured from these hosts indicates the existence of low density or "background"-symbiont populations and cryptic, potentially non-mutualistic types in some hosts.
Palytoxin is a potent marine toxin and one of the most complex natural compounds ever described. A number of compounds identified as palytoxin congeners (e.g., ovatoxins, mascarenotoxins, ostreocins, etc.) have not been yet structurally elucidated due to lack of pure material in quantities sufficient to an NMR-based structural investigation. In this study, the complex fragmentation pattern of palytoxin in its positive high resolution liquid chromatography tandem mass spectra (HR LC-MS(n)) was interpreted. Under the used conditions, the molecule underwent fragmentation at many sites of its backbone, and a large number of diagnostic fragment ions were identified. The natural product itself was used with no need for derivatization. Interestingly, most of the fragments contained calcium in their elemental formula. Evidence for palytoxin tendency to form adduct ions with calcium and other divalent cations in its mass spectra was obtained. Fragmentation pattern of palytoxin was used as template to gain detailed structural information on ovatoxin-a, the main toxin produced by Ostreopsis ovata, (observe correct font) a benthic dinoflagellate that currently represents the major harmful algal bloom threat in the Mediterranean area. Either the regions or the specific sites where ovatoxin-a and palytoxin structurally differ have been identified.
Marine pelagic diazotrophic cyanobacteria of the genus Trichodesmium (Oscillatoriales) are widespread throughout the tropics and subtropics, and are particularly common in the waters of New Caledonia. Blooms of Trichodesmium are suspected to be a potential source of toxins in the ciguatera food chain and were previously reported to contain several types of paralyzing toxins. The toxicity of water-soluble extracts of Trichodesmium spp. were analyzed by mouse bioassay and Neuroblastoma assay and their toxic compounds characterized using liquid chromatography coupled with tandem mass spectrometry techniques. Here, we report the first identification of palytoxin and one of its derivatives, 42-hydroxy-palytoxin, in field samples of Trichodesmium collected in the New Caledonian lagoon. The possible role played by Trichodesmium blooms in the development of clupeotoxism, this human intoxication following the ingestion of plankton-eating fish and classically associated with Ostreopsis blooms, is also discussed.
Zoanthids (Anthozoa, Hexacorallia) are colonial anemones that contain one of the deadliest toxins ever discovered, palytoxin (LD(50) in mice 300 ng/kg), but it is generally believed that highly toxic species are not sold in the home aquarium trade. We previously showed that an unintentionally introduced zoanthid in a home aquarium contained high concentrations of palytoxin and was likely responsible for a severe respiratory reaction when an individual attempted to eliminate the contaminant colonies using boiling water. To assess the availability and potential exposure of palytoxin to marine aquarium hobbyists, we analyzed zoanthid samples collected from local aquarium stores for palytoxin using liquid chromatography and high resolution mass spectrometry and attempted to identify the specimens through genetic analysis of 16S and cytochrome c oxidase 1 (COI) markers. We found four specimens of the same apparent species of zoanthid, that we described previously to be responsible for a severe respiratory reaction in a home aquarium, to be available in three aquarium stores in the Washington D.C. area. We found all of these specimens (n = 4) to be highly toxic with palytoxin or palytoxin-like compounds (range 0.5-3.5 mg crude toxin/g zoanthid). One of the most potent non-protein compounds ever discovered is present in dangerous quantities in a select species of zoanthid commonly sold in the home aquarium trade.
This paper gives descriptions of zoanthids collected from coral reefs and shores of the Hawaiian Islands between June 1965 and June 1967. Isaurus elongatusVerrill (1928) and Palythoa tuberculosaEsper (1791) are redescribed and Zoanthus vestitusVerrill (1928) is reclassified as Palythoa vestitus. Zoanthus confertusVerrill (1928) and Z. nitidusVerrill (1928) are combined under the new species name Zoanthus pacificus. New species described are P. psammophilia, P. toxica, and Z. kealakekuaensis. A list of zoanthid species from the south-western Pacific Ocean is given.
Describes a marine toxin found in Palythoa toxica, a 'soft coral', occurring in a single small tidepool. Toxicity and pharmacological properties, and its potential use in studying diseases of the heart and possibly in cancer research, are outlined. -J.Harvey
This study provides the first evaluation of the cytotoxic effects of the recently-identified palytoxin (PLTX) analog, ovatoxin-a (OVTX-a), the major toxin produced by Ostreopsis cf. ovata in the Mediterranean Sea. Its increasing detection during Ostreopsis blooms and in seafood highlights the need to characterize its toxic effects and to set up appropriate detection methods. OVTX-a is about 100 fold less potent than PLTX in reducing HaCaT cells viability (EC50=1.1x10-9 M vs 1.8x10-11 M, MTT test) in agreement with a reduced binding affinity (Kd=1.2x10-9 vs 2.7x10-11 M, saturation experiments on intact cells). Similarly, OVTX-a hemolytic effect is lower than that of the reference PLTX compound. Ost-D shows the lowest cytotoxicity toward HaCaT keratinocytes, suggesting the lack of a hydroxyl group at C44 as a critical feature for PLTXs cytotoxic effects. A sandwich ELISA developed for PLTX detects also OVTX-a in a sensitive (LOD=4.2 and LOQ=5.6 ng/ml) and accurate manner (Bias=0.3%), also in O. cf. ovata extracts and contaminated mussels. Although in vitro OVTX-a appears less toxic than PLTX, its cytotoxicity at nanomolar concentrations after short exposure time rises some concern for human health. The sandwich ELISA can be a viable screening method for OVTXs detection in monitoring program.
Anecdotal reports exist of aquarium hobbyists that experienced severe respiratory distress and/or skin injury following cleaning operation of home aquaria containing Palythoa sp. soft corals. Hundreds of cases of respiratory illness and/or dermatitis have been recorded in proximity to the sea concomitantly with algal blooms of Ostreopsis spp. in the Mediterranean area. Both Palythoa spp. and Ostreopsis spp. contain congeners of palytoxin, a highly potent toxin whose inhalation hazard is however unknown. In this study, we demonstrate the presence of high levels of palytoxins (palytoxin and hydroxypalytoxin) in both soft coral and seawater from a home marine aquarium involved in the poisoning of a whole family. Due to the high toxin levels found in seawater, a procedure for a rapid and efficient determination of palytoxin in seawater was set-up. A comparison of symptoms of Palythoa- and Ostreopsis-related inhalatory poisonings showed many similarities including fever, respiratory distress, nausea, and flu-like symptoms. From the chemical and symptomatological data reported herein it is reasonable to hold palytoxins responsible for respiratory disorders following inhalation. Although the exact mechanism through which palytoxin congeners exert their inhalatory toxicity is still unknown, this represents a step toward demonstrating that palytoxin congeners exert toxic effects through inhalation both in natural environments and in the surroundings of private and public aquaria.
Although frequently observed in domestic saltwater aquariums, literature on exposure to palytoxin (PTX) of encrusting anemones (Zoanthidea) kept in aquariums is rare. Handling these animals for propagation purposes or during cleaning work can lead to dermal, ocular or respiratory contact with the PTX generated by some Zoanthids. The present study describes a case of ocular exposure to liquid from a Zoanthid, which led to corneal ulcers. The patient also suffered from systemic symptoms of dyspnea and shivering and a suspected rhabdomyolysis, which required monitoring in the Intensive Care Unit. After symptomatic treatment provided insufficient results, the corneal ulcers improved with an amniotic membrane transplantation. A review of the literature regarding ocular exposures to this diverse order of Hexacorallia reveals that severe and systemic symptoms can develop with minimal contact.
On August 12, 2014, an Anchorage hospital notified the Alaska Section of Epidemiology (SOE) that a middle-aged male resident of Anchorage (patient A) had arrived in the emergency department with possible palytoxin exposure. Patient A complained of a bitter metallic taste, fever, weakness, cough, and muscle pain 7-8 hours after introduction of live zoanthid coral into his home aquarium. Palytoxin, a potent toxin known to produce the reported effects, is contained in zoanthid marine corals.
This study is the first review of Zoantharia species in the South China Sea and Gulf of Thailand. In addition to past literature records, new field observations are added from previously unexamined countries and regions. In total 16 species are listed, 15 of which belong to suborder Brachycnemina, and only one to suborder Macrocnemina. Two species are undescribed. The lack of Macrocnemina species is not likely indicative of a low diversity of this suborder in the South China Sea and Gulf of Thailand, but instead of an absence of research below shallow subtidal depths. As the majority of the new records from this study were randomly compiled by researchers who are not experts of Zoantharia, specific surveys by experts are needed in these two marine regions. The present list should provide a solid basis for such future research.
The presence and diversity of free-living dinoflagellates belonging to the endosymbiotic genus Symbiodinium were explored in seawater samples collected above coral reefs in Kāne'ohe Bay, O'ahu, Hawai'i, and Puerto Morelos, Quintana Roo, Mexico. Two genetic markers were used to assess Symbiodinium diversity in the water column: the internal transcribed spacer 2 region of the nuclear ribosomal array (ITS2), and a hypervariable region in domain V of the large subunit (23S) of the chloroplast ribosomal array (cp23S-HVR). Sequencing of cloned gene fragments reveals that clades B, C, D, and H Symbiodinium are detectable in the seawater samples. In addition to the previously described types Bl, C3, C15, C21, and D1, novel Symbiodinium sequences belonging to clades B and C were also retrieved. The majority of Symbiodinium sequences recovered from Kāne'ohe Bay belonged to clade C and those from Puerto Morelos to clade B, a pattern that reflects the dominant types of Symbiodinium found in endosymbiosis with scleractinian corals in these two areas. This study represents the first direct assessment of Symbiodinium diversity in waters adjacent to coral reefs located in the Caribbean and the Pacific and confirms the presence of Symbiodinium in this compartment of the ecosystem. These data provide context for future studies examining spatial and temporal patterns in the availability of Symbiodinium in the water column, work that will ultimately promote a greater understanding of the interactions between symbiotic dinoflagellates and their environmentally sensitive benthic hosts.
Harmful Algal Blooms (HABs) are massive proliferations of toxic algal species that occur under appropriate climatic and environmental conditions. HABs can exert their adverse impacts both on the environment and on particular living organisms through the production of marine biotoxins. Consumption of contaminated seafood or direct exposure to marine biotoxins (swimming, aerosols etc.) can result in human illness or even death. Thousands of cases of marine-toxin-related human poisoning are reported every year on a global basis. In Italy, research on marine biotoxins has shown a continuously changing toxin profile, with most of the toxin classes known so far being found at some point in seafood and seawater. Starting in the late 1980s, with the first finding of a marine biotoxin, okadaic acid, along the Italian coastline, a number of other classes of marine toxins – namely, yessotoxins, pectenotoxins, saxitoxins, domoic acid, spirolides, palytoxins and azaspiracids – have been found. Here we provide an overview, in chronological order, of the many classes of marine biotoxins that have infested Italian seawater over the past 25 years.
Palytoxin ranks among the most potent marine biotoxins. Its lethality was well known to native Hawaiians that used to smear a "moss" containing the toxin on their spears to cause instant death to their victims. Human intoxications due to exposure to palytoxin and to its many congeners have been reported worldwide. Currently, palytoxins constitute the main threat to public health across the Mediterranean Sea. In the present work we report on the isolation and stereostructural determination of a new palytoxin analogue from a Hawaiian Palythoa tuberculosa sample. This new toxin is a stereoisomer of 42-hydroxypalytoxin isolated from Palythoa toxica. The whole absolute configuration of this latter toxin is also reported in the paper. Interestingly, the two 42-hydroxypalytoxins do not share the same biological activity. The stereoisomer from P. tuberculosa showed cytotoxicity toward skin HaCaT keratinocytes approximately 1 order of magnitude lower than that of 42-hydroxypalytoxin from P. toxica and about 2 orders of magnitude lower than that of palytoxin itself. This finding holds the prospect of interesting structure-activity relationship evaluations in the future.
The complete structure of palytoxin (1) was elucidated by us in 1982.1 Our continuous interests in palytoxin led us to examine minor constituents of Okinawan Palythoa tuberculosa. In this paper, we describe successful isolation and structural elucidation of four minor toxins, which were named homopalytoxin (2), bishomopalytoxin (3), neopalytoxin (4) and deoxypalytoxin (5).
Shallow water zooxanthellate zoanthids are a common component of the coral reef ecosystems of the Caribbean. Despite this, their species diversity remains poorly understood. In this study, collected Palythoa, Zoanthus, Isaurus, and Terrazoanthus specimens from the waters of Florida were phylogenetically examined to obtain a better understanding of zoanthid species diversity in the Caribbean. Surprisingly, the results from analyses utilizing three DNA markers (mitochondrial 16S ribosomal DNA, cytochrome oxidase subunit I, and the internal transcribed spacer of ribosomal DNA) showed the presence of at least eleven species, of which up to four appear undescribed. Additionally, the presence of the genus Terrazoanthus in the Caribbean was confirmed for the first time. Attempts to match phylogenetic species or clades with original literature were hampered by vague and short original descriptions, and it is clear that for Atlantic Palythoa and Zoanthus species an in-depth and multidisciplinary investigation is needed to reconcile recent phylogenetic results such as in this study with traditional taxonomy. Furthermore, most shallow water zoanthid species from Florida were observed to have close, sister-species relationships with previously investigated species in the Pacific Ocean. These results indicate that many brachycnemic zoanthid species likely had a Caribbean-Pacific distribution until the formation of the Isthmus of Panama. However, due to inadvertent redescriptions, overall species diversity in these two common genera is likely much lower than literature indicates.
The ovatoxins are palytoxin analogs of a dinoflagellate origin implicated in human intoxication. The structures of ovatoxin-a, ovatoxin-d, and ovatoxin-e produced by the IK2 strain of Ostreopsis ovata collected in Japan were elucidated using liquid chromatography/quadrupole time-of-flight mass spectrometry (LC/QTOFMS). The novel structures and a new insight into the spectral data are presented.
The structural elucidations were carried out by complementary use of positive and negative ion LC/QTOFMS. Ostreocin-D (C127 H219 N3 O53 ), another palytoxin congener previously elucidated by negative fast-atom bombardment collision-induced tandem mass spectrometry (FAB CID MS/MS), was used as a reference.
Positive ion spectra allowed deduction of hydroxyl positions based on the conjugated polyene structures produced, while the negative ion spectra allowed assignments of cleavage sites of C-C bonds. The analysis could be performed using a small sample without extensive purification.
Ovatoxin-a IK2 (C129 H223 N3 O52 ), ovatoxin-d IK2 (C129 H223 N3 O53 ), and ovatoxin-e IK2 (C129 H223 N3 O53 ) were tentatively assigned to 42-hydroxy-17,44,70-trideoxypalytoxin, 42-hydroxy-17,70-dideoxypalytoxin and 42,82-dihydroxy-17,44,70-trideoxypalytoxin, respectively. The wide applicability of the method was suggested. Copyright © 2013 John Wiley & Sons, Ltd.
Palytoxin, which is a toxin with a molecular weight of 2681 daltons isolated from a marine coelenterate, is a potent skin
irritant. However, it did not induce ornithine decarboxylase in mouse skin, or adhesion of human promyelocytic leukemia cells
(HL-60). Moreover, it did not inhibit the specific binding of (3H)12-O-tetradecanoylphorbol-13-acetate (TPA) to a mouse skin particulate fraction or activate protein kinase C isolated from mouse
brain in vitro. Since palytoxin showed strong irritation on mouse ear in one short-term screening test for a promoter, it was examined in
a two-stage carcinogenesis experiment. The incidence of tumors in a group of mice treated with 7, 12-dimethylbenz(a)anthracene
plus palytoxin was 62.5% in week 25. These tumors were identified histologically as seven papillonmas and one carcinoma. This
paper reports the potent tumor-promoting activity of palytoxin, which is classified as a non-TPA-type tumor promoter.
Palytoxins are potent marine biotoxins that have recently become endemic to the Mediterranean Sea, and are becoming more frequently associated with seafood. Due to their high toxicity, suitable methods to quantify palytoxins are needed. Thus, we developed an indirect sandwich ELISA for palytoxin and 42-hydroxy-palytoxin. An intra-laboratory study demonstrated sensitivity (limit of detection, LOD=1.1 ng/ml; limit of quantitation, LOQ=2.2 ng/ml), accuracy (bias of 2.1%), repeatability (RSDr=6% and 9% for intra- and inter-assay variability, respectively) and specificity: other common marine toxins (okadaic acid, domoic acid, saxitoxin, brevetoxin-3 and yessotoxin) don't cross-react in this assay. It performed well in three different matrices: observed LOQs were 11.0, 9.6, and 2.4 ng/ml for mussel extracts, algal net samples and seawater, respectively, with good accuracy and precision. The LOQ in seafood is 11 µg palytoxin/kg mussel meat, lower than that of the most common detection technique, LC-MS/MS.
In the last decades, massive blooms of palytoxin (PLTX)-producing Ostreopsis cf. ovata have been observed along Mediterranean coasts, usually associated to human respiratory and cutaneous problems. At the molecular level, PLTX induces a massive intracellular Na(+) influx due to the transformation of Na(+)/K(+) ATPase in a cationic channel. Recently, we have demonstrated that Na(+) overload is the crucial step in mediating overproduction of reactive oxygen species (ROS) and cell death in human HaCaT keratinocytes, tentatively explaining PLTX-induced skin irritant effects. In the present study the molecular mechanisms of ROS production induced by PLTX-mediated Na(+) intracellular overload have been investigated. In HaCaT cells, PLTX exposure caused accumulation of superoxide anion, but not of nitric oxide or peroxynitrite/hydroxyl radicals. Even if RT-PCR and western blot analysis revealed an early NOX-2 and iNOS gene and protein over-expressions, their active involvement seemed to be only partial since selective inhibitors did not completely reduce O(2)(-) production. A significant role of other enzymes (COX-1, COX-2, XO) was not evidenced. Nigericin, that counteracts Na(+)-mediated H(+)-imbalance, dissipating ΔpH across mitochondrial inner membrane, and the uncouplers DNP significantly reduced O(2)(-) production. These inhibitions were synergistic when co-exposed with complex-I inhibitor rotenone. These results suggest a novel mechanism of O(2)(-) production induced by PLTX-mediated ionic imbalance. Indeed, the H(+) intracellular overload that follows PLTX-induced intracellular Na(+) accumulation, could enhance ΔpH across mitochondrial inner membrane, that seems to be the driving force for O(2)(-) production by reversing mitochondrial electron transport.
This paper reports on the analysis of the toxin content from Palythoa tuberculosa and Palythoa toxica samples collected off of the Hawaiian coast. Our work, based on in-depth high-resolution liquid chromatography−mass spectrometry analysis along with extensive NMR study, led us to structurally characterize 42-hydroxy-palytoxin, a new palytoxin congener. This toxin and palytoxin itself appeared to be the major components of toxic extract from a P. tuberculosa sample, while 42-hydroxy-palytoxin was proven by far to be the main palytoxin derivative in P. toxica. Functional studies on this new palytoxin-like compound suggest that the new palytoxin analogue and palytoxin itself present similar biological activities.
This paper gives descriptions of zoanthids collected from coral reefs and shores of the Hawaiian Islands between June 1965 and June 1967. Isaurus elongatus Verrill (1928) and Palythoa tuberculosa Esper (1791) are redescribed and Zoanthus vestitus Verrill (1928) is reclassified as Palythoa vestitus. Zoanthus confertus Verrill (1928) and Z. nitidus Verrill (1928) are combined under the new species name Zoanthus paciflcus. New species described are P. psammophilia, P. toxica, and Z. kealakehuaensis. A list of zoanthid species from the south-western Pacific Ocean is given.
No abstract available.
Previous phylogenetic studies based on mitochondrial DNA markers have suggested that the zoanthid genus Palythoa may consist of both Palythoa species (Palythoa tuberculosa) and species formerly assigned to the genus Protopalythoa (Palythoa mutuki, Palythoa heliodiscus). In the present study various Palythoa spp. samples collected primarily from southern Japan with additional samples from the Indo-Pacific and Caribbean Sea were
examined. The nuclear internal transcribed spacer of ribosomal DNA (ITS-rDNA) was sequenced and aligned for phylogenetic analyses
to further investigate the relationship between P. tuberculosa, P. mutuki, and P. heliodiscus. ITS-rDNA analyses showed species groups forming monophylies with similar topology but with much higher resolution than seen
for mitochondrial phylogenetic analyses. The results also confirmed the very close relationship of P. tuberculosa and P. mutuki. Some specimens appeared to be a potentially undescribed Palythoa species (designated Palythoa sp. sakurajimensis). Additionally, ITS-rDNA sequences of P. mutuki and P. tuberculosa showed additive polymorphic site, demonstrating for the first time a potential history of reticulate evolution in Palythoa.
Palytoxin (PTX) was first isolated from the zoanthid Palythoa toxica. Evaluation of PTX toxicity using various animal models determined that PTX was extremely potent through intravenous, intraperitoneal, and intratracheal exposure. PTX was less potent by direct intragastric exposure. PTX also caused significant, non-lethal effects through dermal and ocular exposure. PTX and PTX-like compounds have now been found in additional zoanthid species, red alga, a sea anemone, and several dinoflagellates. PTXs are found throughout certain reef associated food webs, including in fish and crabs responsible for human illness and death. Many of the organisms found to contain PTXs in the environment are also sold in the home aquarium trade, and recent evidence suggests poisonings have occurred through exposure to these organisms. Due to co-occurrence with other seafood toxins, such as ciguatoxins, saxitoxins, and tetrodotoxin, it has been difficult to assess the true risk of PTX poisoning through seafood consumption in humans, but limited cases have been well documented, some involving human fatalities. Recent evidence also suggests that humans are negatively impacted through PTX exposure by inhalation and dermal routes. Continued research into the distribution and occurrence of PTX and PTX-like compounds both in seafood and marine organisms sold in the aquarium trade appears warranted.
Improved aquarium techniques are enabling sea aquariums to imitate reality even better. This means that they sometimes contain known and unknown marine toxins. Aquarium keepers and their families may then develop potentially severe poisoning.
A 37-year-old man presented at the Emergency Department with fever, hypotension and symptoms of a metallic taste in the mouth, nausea, headache, shivering and severe muscle cramps. Symptoms appeared after he had tried to remove a colony of Zoanthids (soft corals) from his aquarium by pouring boiling water over them. His wife (35-year-old) and two children (10-year-old twins) had also inhaled the offensive-smelling steam coming from the aquarium and had similar symptoms. On physical examination, all family members had low blood pressure and fever > 38.5 degrees Celsius. Blood analysis showed leucocytosis and an elevated CRP. We diagnosed palytoxin poisoning. All family members recovered within 48 hours after receiving supportive therapy only.
Manipulation of certain soft corals found in sea aquariums can cause emission of palytoxin. This may cause a potentially severe episode of poisoning. When working in a sea aquarium protective measures should be taken. There is no specific therapy for or antidote to palytoxin poisoning and for this reason, treatment is supportive.
A series of case reports and anecdotal references describe the adverse effects on human health ascribed to the marine toxin palytoxin (PLTX) after different exposure routes. They include poisonings after oral intake of contaminated seafood, but also inhalation and cutaneous/systemic exposures after direct contact with aerosolized seawater during Ostreopsis blooms and/or through maintaining aquaria containing cnidarian zoanthids. The symptoms commonly recorded during PLTX intoxication are general malaise and weakness, associated with myalgia, respiratory effects, impairment of the neuromuscular apparatus and abnormalities in cardiac function. Systemic symptoms are often recorded together with local damages whose intensity varies according to the route and length of exposure. Gastrointestinal malaise or respiratory distress is common for oral and inhalational exposure, respectively. In addition, irritant properties of PLTX probably account for the inflammatory reactions typical of cutaneous and inhalational contact. Unfortunately, the toxin identification and/or quantification are often incomplete or missing and cases of poisoning are indirectly ascribed to PLTXs, according only to symptoms, anamnesis and environmental/epidemiological investigations (i.e. zoanthid handling or ingestion of particular seafood). Based on the available literature, we suggest a "case definition of PLTX poisonings" according to the main exposure routes, and, we propose the main symptoms to be checked, as well as, hemato-clinical analysis to be carried out. We also suggest the performance of specific analyses both on biological specimens of patients, as well as, on the contaminated materials responsible for the poisoning. A standardized protocol for data collection could provide a more rapid and reliable diagnosis of palytoxin-poisoning, but also the collection of necessary data for the risk assessment for this family of toxins.
Palytoxin (PLTX), a marine toxin identified in Palythoa zoanthid corals and Ostreopsis dinoflagellates, represents an increasing hazard for human health. Recently, dermatological problems have been associated to cutaneous exposure to PLTX during Ostreopsis blooms arising the need for experimental data characterizing PLTX effects on the skin. This study highlights in vitro the cytotoxic effects of PLTX on human keratinocytes (HaCaT cell line). A short time exposure (4h) to PLTX reduced mitochondrial activity (MTT assay), cell mass (SRB assay) and plasma membrane integrity (LDH leakage) with different potencies: EC₅₀ values of 6.1 ± 1.3×10⁻¹¹, 4.7 ± 0.9 × 10⁻¹⁰ M and 1.8 ± 0.1 × 10⁻⁸ M, respectively. PLTX effect on mitochondrial activity was ouabain- and Na+-sensitive, but only partially sensitive to removal of Ca²+ ions. One hour exposure to the toxin also induced a Na+-dependent and Ca²+-independent superoxide anion production. These results indicate that among the chain of intracellular events following the interaction of PLTX with the Na+/K+-ATPase the first and crucial step is the increased intracellular Na+ concentration that triggers a sequence of cell dysfunction involving mitochondrial affection and oxidative stress, leading to an irreversible cell death. The PLTX concentrations inducing cytotoxicty seem to be lower than those of potential cutaneous human exposure during Ostreopsis ovata blooms, indicating the harmful potential of the toxin.