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1.
PLoS Negl Trop Dis ; 14(3): e0008060, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32163415

RESUMO

The northeast (NE) region of Brazil commonly goes through drought periods, which favor cyanobacterial blooms, capable of producing neurotoxins with implications for human and animal health. The most severe dry spell in the history of Brazil occurred between 2012 and 2016. Coincidently, the highest incidence of microcephaly associated with the Zika virus (ZIKV) outbreak took place in the NE region of Brazil during the same years. In this work, we tested the hypothesis that saxitoxin (STX), a neurotoxin produced in South America by the freshwater cyanobacteria Raphidiopsis raciborskii, could have contributed to the most severe Congenital Zika Syndrome (CZS) profile described worldwide. Quality surveillance showed higher cyanobacteria amounts and STX occurrence in human drinking water supplies of NE compared to other regions of Brazil. Experimentally, we described that STX doubled the quantity of ZIKV-induced neural cell death in progenitor areas of human brain organoids, while the chronic ingestion of water contaminated with STX before and during gestation caused brain abnormalities in offspring of ZIKV-infected immunocompetent C57BL/6J mice. Our data indicate that saxitoxin-producing cyanobacteria is overspread in water reservoirs of the NE and might have acted as a co-insult to ZIKV infection in Brazil. These results raise a public health concern regarding the consequences of arbovirus outbreaks happening in areas with droughts and/or frequent freshwater cyanobacterial blooms.


Assuntos
Morte Celular/efeitos dos fármacos , Microcefalia/patologia , Envenenamento/complicações , Envenenamento/patologia , Saxitoxina/toxicidade , Infecção por Zika virus/complicações , Infecção por Zika virus/patologia , Animais , Toxinas Bacterianas/análise , Toxinas Bacterianas/toxicidade , Encéfalo/patologia , Brasil/epidemiologia , Células Cultivadas , Modelos Animais de Doenças , Surtos de Doenças , Feminino , Humanos , Incidência , Toxinas Marinhas/análise , Toxinas Marinhas/toxicidade , Camundongos Endogâmicos C57BL , Microcistinas/análise , Microcistinas/toxicidade , Modelos Teóricos , Neurotoxinas/análise , Neurotoxinas/toxicidade , Saxitoxina/análise , Água/química
2.
Artigo em Inglês | MEDLINE | ID: mdl-31638869

RESUMO

There are two official PSP detection methods (mouse bioassay and HLPC-FLD) and a number of alternative methods. Ethical considerations have led to regulations being adopted in some countries that limit or prohibit the application of mouse bioassay. Analytical methodologies (e.g. HPLC-FLD or LC-MSMS) have the disadvantages of not being able to detect new toxins or analogues or reflecting the overall toxicity of the sample. In addition, they require highly trained personnel and expensive equipment, which are not always available. In this work, we have evaluated a method based on the Neuro-2a cell-based assay to detect substances that inhibit voltage-dependent sodium channels (Manger's method). We tested PSP standards and natural samples contaminated with PSP. Here we demonstrate that the adapted Manger's method is suitable for calculating Toxicity Equivalency Factors (TEF) for STX-analogues. The method was shown to be useful for screening contaminated natural samples in concentrations above the regulatory limit for these toxins (80 µg STX equivalents/100 g shellfish). We were able to detect PSP in 19 natural mollusc samples from South Chile despite the presence of other marine toxins. These preliminary results suggest that the method could be used as a first step in screening programmes.


Assuntos
Análise de Alimentos , Contaminação de Alimentos/análise , Saxitoxina/análise , Saxitoxina/toxicidade , Alimentos Marinhos/análise , Alimentos Marinhos/toxicidade , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Chile , Relação Dose-Resposta a Droga , Camundongos , Frutos do Mar , Intoxicação por Frutos do Mar
3.
Aquat Toxicol ; 218: 105371, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31790939

RESUMO

Harmful algal bloom (HAB) toxins have severe negative impacts on marine mammals, particularly for Florida bottlenose dolphins (Tursiops truncatus) which frequently experience mass mortality events. Dolphins on the Florida Atlantic coast inhabit a region endemic to two HAB species, Karenia brevis and Pyrodinium bahamense, which produce the neurotoxins brevetoxin (PbTx) and saxitoxin (STX), respectively. Although toxic HABs and associated dolphin mortality events have been reported from this region, there is a lack of available data necessary for comparing toxin exposure levels between bloom ('exposed') conditions and non-bloom ('baseline') conditions. Here we present a 10-year dataset of PbTx and STX concentrations detected in dolphins stranding in this region, and compare the toxin loads from HAB-exposed dolphins to those detected in dolphins recovered in the absence of a HAB. We analyzed liver tissue samples from dead-stranded dolphins (n = 119) recovered and necropsied between 2002-2011, using an enzyme-linked immunosorbent assay (ELISA) modified for use with mammalian tissues. For dolphins recovered during baseline conditions, toxin-positive samples ranged in concentration from 0.27 to 1.2 ng/g for PbTx and from 0.41 to 1.9 ng/g for STX. For K. brevis-exposed dolphins, concentrations of up to 12.1 ng PbTx/g were detected, and for P. bahamense-exposed dolphins, concentrations of up to 9.9 ng STX/g were detected. Baseline PbTx values were similar to those reported in other regions where K. brevis blooms are more frequent and severe, but HAB-exposed PbTx values were considerably lower relative to these other regions. Since no baseline STX dolphin data exist for any region, our data serve as a first step towards establishing reference STX values for potential dolphin mortality events associated with STX-producing blooms in the future. This study demonstrates that although HABs in eastern Florida are only infrequently associated with dolphin mortalities, the presence of toxins in these animals may pose significant health risks in this region.


Assuntos
Golfinho Nariz-de-Garrafa/metabolismo , Monitoramento Ambiental/métodos , Proliferação Nociva de Algas , Toxinas Marinhas/análise , Oxocinas/análise , Saxitoxina/análise , Poluentes Químicos da Água/análise , Animais , Dinoflagelados/crescimento & desenvolvimento , Ensaio de Imunoadsorção Enzimática , Florida , Fígado/química , Fígado/metabolismo , Toxinas Marinhas/toxicidade , Oxocinas/toxicidade , Rios/química , Saxitoxina/toxicidade , Poluentes Químicos da Água/toxicidade
4.
Mar Drugs ; 17(12)2019 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-31766477

RESUMO

Paralytic shellfish toxins (PSTs) are the major neurotoxic contaminants of edible bivalves in Japan. Tetrodotoxin (TTX) was recently detected in bivalve shellfish around the world, drawing widespread attention. In Japan, high levels of TTX were reported in the digestive gland of the scallop, Patinopecten yessoensis, in 1993; however, no new data have emerged since then. In this study, we simultaneously analyzed PSTs and TTX in scallops cultured in a bay of east Japan using hydrophilic interaction chromatography (HILIC)-MS/MS. These scallops were temporally collected from April to December 2017. The highest concentration of PSTs (182 µmol/kg, total congeners) in the hepatopancreas was detected in samples collected on May 23, lined to the cell density of the dinoflagellate, Alexandrium tamarense, in seawater around the scallops, whereas the highest concentration of TTX (421 nmol/kg) was detected in samples collected on August 22. Contrary to the previous report, temporal variation of the PSTs and TTX concentrations did not coincide. The highest concentration of TTX in the entire edible tissues was 7.3 µg/kg (23 nmol/kg) in samples obtained on August 22, which was lower than the European Food Safety Authority (EFSA)-proposed threshold, 44 µg TTX equivalents/kg shellfish meat. In addition, 12ß-deoxygonyautoxin 3 was firstly identified in scallops.


Assuntos
Dinoflagelados/química , Pectinidae/química , Saxitoxina/análogos & derivados , Alimentos Marinhos/análise , Tetrodotoxina/análise , Animais , Aquicultura , Baías , Cromatografia Líquida de Alta Pressão , Japão , Saxitoxina/análise , Saxitoxina/toxicidade , Estações do Ano , Água do Mar/microbiologia , Intoxicação por Frutos do Mar/etiologia , Intoxicação por Frutos do Mar/prevenção & controle , Espectrometria de Massas em Tandem , Tetrodotoxina/toxicidade , Fatores de Tempo
5.
Toxicon ; 171: 62-65, 2019 Dec 05.
Artigo em Inglês | MEDLINE | ID: mdl-31614156

RESUMO

Algal toxins (domoic acid, saxitoxin, okadaic acid) were monitored at seven locations off southern California using Solid Phase Adsorption Toxin Tracking. At least two types of toxins were found at all locations, with co-occurrence of two and three toxins in 12% and 10% of samples, respectively. This study expands our limited understanding of the simultaneous presence of multiple algal toxins along the coast and raises questions regarding the potential health ramifications of such co-occurrences.


Assuntos
Ácido Caínico/análogos & derivados , Ácido Okadáico/análise , Saxitoxina/análise , Adsorção , California , Monitoramento Ambiental , Proliferação Nociva de Algas , Ácido Caínico/análise , Toxinas Marinhas/análise , Água do Mar/química
6.
Chemosphere ; 236: 124404, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31545201

RESUMO

Tetrodotoxin (TTX) is a potent neurotoxin responsible for countless human intoxications and deaths around the world. The distribution of TTX and its analogues is diverse and the toxin has been detected in organisms from both marine and terrestrial environments. Increasing detections seafood species, such as bivalves and gastropods, has drawn attention to the toxin, reinvigorating scientific interest and regulatory concerns. There have been reports of TTX in 21 species of bivalves and edible gastropods from ten countries since the 1980's. While TTX is structurally dissimilar to saxitoxin (STX), another neurotoxin detected in seafood, it has similar sodium channel blocking action and potency and both neurotoxins have been shown to have additive toxicities. The global regulatory level for the STX group toxins applied to shellfish is 800 µg/kg. The presence of TTX in shellfish is only regulated in one country; The Netherlands, with a regulatory level of 44 µg/kg. Due to the recent interest surrounding TTX in bivalves, the European Food Safety Authority established a panel to assess the risk and regulation of TTX in bivalves, and their final opinion was that a concentration below 44 µg of TTX per kg of shellfish would not result in adverse human effects. In this article, we review current knowledge on worldwide TTX levels in edible gastropods and bivalves over the last four decades, the different methods of detection used, and the current regulatory status. We suggest research needs that will assist with knowledge gaps and ultimately allow development of robust monitoring and management protocols.


Assuntos
Bivalves/química , Contaminação de Alimentos/análise , Gastrópodes/química , Frutos do Mar/análise , Tetrodotoxina/análise , Animais , Contaminação de Alimentos/legislação & jurisprudência , Inocuidade dos Alimentos , Humanos , Países Baixos , Neurotoxinas/análise , Neurotoxinas/farmacocinética , Saxitoxina/análise , Tetrodotoxina/farmacocinética
7.
Toxicon ; 167: 76-81, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31170406

RESUMO

To understand phycotoxin contamination in shellfish in the sub-Arctic and Arctic areas, scanning for the presence of 13 hydrophilic and lipophilic toxin components each was by liquid chromatography tandem quadrupole mass spectrometry analysis in shellfish samples collected from the Northern Bering Sea and the Chukchi Sea in 2014. The results showed that shellfish collected in both areas werecontaminated to different extents. Saxitoxin (STX), decarbamoylsaxitoxin (dcSTX) and decarbamoylneosaxitoxin (dcNEO) were the most frequently detected hydrophilic components, with maximum concentrations of 90.1 µg/kg, 112.25 µg/kg and 23.09 µg/kg, respectively. Although gonyautoxins (GTXs) were only detected in 3 samples, they were the main contributors to overall toxicity of high-latitude samples, especially GTX1. For lipophilic toxins, spirolide-1 (SPX1) and yessotoxin (YTX) were present in all samples at low levels (< 7 µg/kg and < 50 µg/kg, respectively). Only 5 samples showed evidence of okadaic acid (OA) and dinophysistoxin-2 (DTX-2) at low concentrations, ranging from 0.42 µg/kg to 7.23 µg/kg and 3.03 µg/kg to 30.59 µg/kg, respectively. Notably, a high level of pectenotoxin-1 (PTX-1) at 467.40 µg/kg was found in the shellfish collected at the northernmost station, exceeding the safety regulation standard by nearly 3 times. For both lipophilic and hydrophilic toxins, contamination in shellfish in the sub-Arctic and the Arctic area may be much more widespread and severe than was previously thought. This study highlighted the need to monitor toxins in a wider variety of shellfish, especially economic or commercial species, and across a wider range of sub-Arctic and Arctic waters, as well as the potential sources of these toxins.


Assuntos
Contaminação de Alimentos/análise , Saxitoxina/análise , Frutos do Mar , Regiões Árticas , Cromatografia Líquida , Saxitoxina/análogos & derivados , Saxitoxina/química , Espectrometria de Massas em Tandem
8.
Toxins (Basel) ; 11(5)2019 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-31096619

RESUMO

Seafood is an emerging health food, and interest in improving the quality of seafood is increasing. Saxitoxin (STX) is a neurotoxin produced by marine dinoflagellates that is accumulated in seafood. It can block the neuronal transmission between nerves and muscle cell membranes, resulting in the disturbance of neuromuscular transmission and subsequent voluntary muscle paralysis. Here, we developed a new aptamer for the detection of STX using graphene oxide-systematic evolution of ligands by exponential enrichment (GO-SELEX). Furthermore, we confirmed sensitivity and selectivity of the developed aptamer specific to STX using a localized surface plasmon resonance (LSPR) sensor. The sensing chip was fabricated by fixing the new STX aptamer immobilized on the gold nanorod (GNR) substrate. The STX LSPR aptasensor showed a broad, linear detection range from 5 to 10,000 µg/L, with a limit of detection (LOD) of 2.46 µg/L (3σ). Moreover, it was suitable for the detection of STX (10, 100, and 2000 µg/L) in spiked mussel samples and showed a good recovery rate (96.13-116.05%). The results demonstrated that the new STX aptamer-modified GNR chip was sufficiently sensitive and selective to detect STX and can be applied to real samples as well. This LSPR aptasensor is a simple, label-free, cost-effective sensing system with a wide detectable range.


Assuntos
Bivalves/química , Neurotoxinas/análise , Saxitoxina/análise , Animais , Aptâmeros de Nucleotídeos/química , Contaminação de Alimentos/análise , Ouro/química , Grafite/química , Nanotubos/química , Alimentos Marinhos/análise , Ressonância de Plasmônio de Superfície
9.
J Chromatogr A ; 1599: 66-74, 2019 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-30961962

RESUMO

Cyanobacteria can form dense blooms under specific environmental conditions, and some species produce secondary metabolites known as cyanotoxins, which present significant risks to public health and the environment. Identifying toxins produced by cyanobacteria present in surface water and fish is critical to ensuring high quality food and water for consumption, and protectionn of recreational uses. Current analytical screening methods typically focus on one class of cyanotoxins in a single matrix and rarely include saxitoxin. Thus, a cross-class screening method for microcystins, nodularin, anatoxin-a, cylindrospermopsin, and saxitoxin was developed to examine target analytes in environmental water and fish tissue. This was done, due to the broad range of cyanotoxin physicochemical properties, by pairing two extraction and separation techniques to improve isolation and detection. For the first time a zwitterionic hydrophilic interaction liquid chromatography column was evaluated to separate anatoxin-a, cylindrospermopsin, and saxitoxin, demonstrating greater sensitivity for all three compounds over previous techniques. Further, the method for microcystins, nodularin, anatoxin-a, and cylindrospermopsin were validated using isotopically labeled internal standards, again for the first time, resulting in improved compensation for recovery bias and matrix suppression. Optimized extractions for water and fish tissue can be extended to other congeners in the future. These improved separation and isotope dilution techniques are a launching point for more complex, non-targeted analyses, with preliminary targeted screening.


Assuntos
Cromatografia Líquida , Monitoramento Ambiental/métodos , Peixes , Toxinas Marinhas/análise , Espectrometria de Massas em Tandem , Água/química , Animais , Toxinas Bacterianas/análise , Cianobactérias/química , Isótopos/análise , Microcistinas/análise , Peptídeos Cíclicos/análise , Saxitoxina/análise , Tropanos/análise , Uracila/análogos & derivados , Uracila/análise
10.
Toxins (Basel) ; 11(1)2019 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-30650549

RESUMO

Butterfield Lake is a mesotrophic lake in New York State where residents and pets have experienced unexplained health issues. Microseira wollei (basionym Lyngbya wollei) was found at two of 15 sites in Butterfield Lake and analyzed for microcystins, anatoxins, cylindrospermopsins, and paralytic shellfish poisoning toxins (PSTs). Only PSTs and trace levels of anatoxin-a were detected in these samples. This is the first published report of PSTs within a New York State lake. To evaluate the environmental and temporal drivers leading to the observed toxicity, PST content at the two sites was examined in detail. There were distinct differences in the total PST content, filament nutrient, filament chlorophyll, and relationship to environmental drivers between the sites, as well as distinct differences in the total PST content measured using different analytical techniques. A multivariate model containing site, temperature, and filament chlorophyll explained 85% of the variation in PSTs observed over the growing season. This work emphasizes the importance of proper site selection and choice of analytical technique in the development of monitoring programs to protect lake users from the occurrence of benthic cyanobacteria toxins.


Assuntos
Toxinas Bacterianas/análise , Cianobactérias/isolamento & purificação , Lagos/análise , Microcistinas/análise , Saxitoxina/análise , Tropanos/análise , Uracila/análogos & derivados , Poluentes da Água/análise , Cianobactérias/genética , Monitoramento Ambiental , New York , RNA Ribossômico 16S , Intoxicação por Frutos do Mar , Uracila/análise
11.
Harmful Algae ; 80: 158-170, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30502809

RESUMO

Strains of the freshwater filamentous, benthic cyanobacterium Scytonema crispum Agardh isolated from six sites in subtropical south-east Queensland were characterised using a combination of phenotypic and genetic traits. Morphologically, the strains were consistent with the description of Scytonemataceae sensu stricto, and the description of Scytonema crispum. However, phylogenetic analysis of the 16S rRNA gene, the 16S-23S rRNA operon, and the nifH gene revealed that these strains and three others from outside Australia formed a monophyletic clade distinct from Scytonema and other species in the Scytonemataceae. Collectively, this data suggests this group is sufficiently evolutionarily distinct to be placed in a new family, Heteroscytonemataceae fam. nov. Accordingly, the taxon previously known as S. crispum has been transferred to a new genus Heteroscytonema gen nov., as H. crispum. Some strains of H. crispum exhibited facultative production of paralytic shellfish toxins (PSTs). The concentration of PSTs produced by individual strains varied widely, from 2.7 µg g-1 to 171.3 µg g-1, and included C toxins, decarbamoyl saxitoxin (dcSTX), gonyautoxins (GTX2, GTX3 and GTX5), saxitoxin (STX) and uncharacterised PSTs. The majority of the Australian strains produced dcSTX as the dominant saxitoxin analogue, a significant finding given that dcSTX has approximately half the relative toxicity of STX. The PST profile varied within and between Australian strains of H. crispum and in strains collected from New Zealand and the United States. The sxtA gene, one of the determinants for the production of PSTs, was present in all strains in which PSTs were detected. The discovery of PST-producing H. crispum in the headwaters of a major drinking water reservoir presents a serious risk for potential human and animal exposure to these neurotoxic compounds and further highlights the importance of monitoring benthic cyanobacteria populations for potentially toxigenic species.


Assuntos
Toxinas Bacterianas/metabolismo , Cianobactérias/classificação , Toxinas Marinhas/metabolismo , Microcistinas/metabolismo , Filogenia , Biodiversidade , Cianobactérias/metabolismo , Queensland , RNA Ribossômico 16S/química , Saxitoxina/análise , Saxitoxina/química , Análise de Sequência de DNA
12.
Toxins (Basel) ; 10(10)2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-30275378

RESUMO

An area of discolored water 50 m wide and 30 m long was found in September 2017 close to the dam of the Irkutsk hydroelectric power station. Water from this spot was sampled for investigation in the present study. Microscopic analysis revealed that the suspended matter in the sample was composed of clumps of filaments, vegetative cells, akinetes and heterocysts that formed short filaments and solitary cells. This matter was found to consist of partially degraded cells of the cyanobacterium Dolichospermum lemmermannii. Nucleotide sequencing of DNA isolated from the biomass revealed the presence of the sxtA gene which is involved in the synthesis of saxitoxin. Water from the polluted area contained 600 ± 100 µg L-1 saxitoxin as measured by HPLC-MS with pre-column modification of the toxin with 2,4-dinitrophenylhydrazine. Immunoassay analysis (ELISA) showed a concentration of saxitoxins in the water of 2900 ± 900 µg L-1. Hydrochemical and microbiological analyses suggested the contaminated area appeared as a result of a D. lemmermannii bloom, followed by its decay and release of saxitoxin and nutrients. The present paper describes the results of a case study. Better understanding of the phenomenon will depend on the possibility to perform implementation of a large-scale monitoring program.


Assuntos
Lagos/química , Saxitoxina/análise , Poluentes Químicos da Água/análise , Abastecimento de Água , Cianobactérias/genética , Cianobactérias/isolamento & purificação , DNA Bacteriano/análise , Monitoramento Ambiental , Lagos/microbiologia , Centrais Elétricas , Saxitoxina/genética , Sibéria
13.
Mar Pollut Bull ; 135: 1198-1204, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30301019

RESUMO

The North Yellow Sea is a major aquaculture production area for the scallop Patinopecten yessoensis. In this study, the temporal and spatial variation of phycotoxins in scallops, phytoplankton, and their cysts were analyzed during a survey conducted from June 2011 to April 2012 around Zhangzi Island. The study area is a semi-enclosed epicontinental sea surrounded by the Shandong Peninsula, the Liaodong Peninsula and the Korean Peninsula. The three main results of the study were as follows: (1) The saxitoxin-group toxins, okadaic acid and analogues, and pectenotoxins were the major phycotoxin residues found in scallops; (2) Six kinds of toxic microalgae were identified, Protoperidinium spp., Gonyaulax spp., and Alexandrium spp. were the dominant taxa; Seven types of potential marine toxin-producing dinoflagellates, A. tamarense, A. catenella, Dinophysis fortii, G. catenatum, Gambierdiscus toxicus, Azadinium poporum, and Pseudo-nitzschia pungen were identified as the primary source of phycotoxins and were present at relatively high density from June to October; and (3) azaspiracids and domoic acid might be new potential sources of toxin pollution. This study represents the first assessment to phycotoxins around Zhangzi Island in the North Yellow Sea.


Assuntos
Contaminação de Alimentos/análise , Toxinas Marinhas/análise , Pectinidae/química , Fitoplâncton , Frutos do Mar/análise , Animais , Aquicultura/métodos , China , Diatomáceas , Dinoflagelados , Microalgas , Oceanos e Mares , Fitoplâncton/química , Saxitoxina/análise , Estações do Ano , Análise Espaço-Temporal , Compostos de Espiro/análise
14.
Toxins (Basel) ; 10(11)2018 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-30373104

RESUMO

Paralytic shellfish poisoning (PSP) is a severe food-borne illness, caused by the ingestion of seafood containing paralytic shellfish toxins (PST), which are naturally produced by marine dinoflagellates and accumulate in shellfish during algae blooms. Novel PST, designated as hydroxybenzoate analogues (also known as GC toxins), was relatively recently discovered in Gymnodinium catenatum strains worldwide. However, to date, there have been no studies examining their accumulation in shellfish. In this study, mussels (Mytilus galloprovincialis) were exposed to G. catenatum for five days and then exposed to a non-toxic diet for 24 h, to investigate the toxin's accumulation/elimination dynamics. As determined by UHPLC-HILIC-MS/MS, the hydroxybenzoate analogues, GC1 to GC6, comprised 41% of the algae toxin profile and only 9% in mussels. Elimination of GC toxins after 24 h was not evident. This study highlights that a relevant fraction of PST in mussels are not routinely analysed in monitoring programs and that there is a need to better understand the toxicological potential of the hydroxybenzoate analogues, in order to properly address the risk of G. catenatum blooms.


Assuntos
Dinoflagelados , Hidroxibenzoatos/análise , Mytilus/metabolismo , Saxitoxina/análogos & derivados , Saxitoxina/análise , Animais , Cromatografia Líquida de Alta Pressão , Hidroxibenzoatos/metabolismo , Saxitoxina/metabolismo , Intoxicação por Frutos do Mar , Espectrometria de Massas em Tandem
15.
Toxins (Basel) ; 10(9)2018 09 06.
Artigo em Inglês | MEDLINE | ID: mdl-30200645

RESUMO

Paralytic shellfish toxins (PSTs) are potent alkaloids of microalgal and cyanobacterial origin, with worldwide distribution. Over the last 20 years, the number of poisoning incidents has declined as a result of the implementation of legislation and monitoring programs based on bivalves. In the summer of 2012 and 2013, we collected a total of 98 samples from 23 different species belonging to benthic and subtidal organisms, such as echinoderms, crustaceans, bivalves, and gastropods. The sampling locations were Madeira, São Miguel Island (Azores archipelago), and the northwestern coast of Morocco. The samples were analyzed using post-column oxidation liquid chromatography with a fluorescence detection method. Our main goal was to detect new vectors for these biotoxins. After reporting a total of 59 positive results for PSTs with 14 new vectors identified, we verified that some of the amounts exceeded the limit value established in the EU. These results suggest that routine monitoring of saxitoxin and its analogs should be extended to more potential vectors other than bivalves, including other edible organisms, for a better protection of public health.


Assuntos
Invertebrados , Saxitoxina/análogos & derivados , Saxitoxina/análise , Poluentes da Água/análise , Animais , Oceano Atlântico , Monitoramento Ambiental , Peixes , Intoxicação por Frutos do Mar
16.
Anal Sci ; 34(8): 893-900, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30101883

RESUMO

Okadaic acid (OA) and saxitoxin (STX) are typical toxins of diarrhetic shellfish poisoning (DSP) and paralytic shellfish poisoning (PSP), respectively, which are highly toxic marine toxins threatening human health and environmental safety. OA is a potent inhibitor of serine/threonine protein phosphatases that can cause cellular death, while STX is an inhibitor of sodium channel that can lead to neurological damage. In this work, a dual functional cardiomyocyte-based biosensor was proposed to detect DSP and PSP toxins by monitoring the viability and electrophysiology of cardiomyocytes. The results showed that the viability of cardiomyocytes was sensitive to the OA and STX, resulting in significant changes of the electrophysiological properties, including amplitude, firing rate and duration of the extracellular field potential (EFP). The detection limits of the hybrid-biosensor are as low as 7.16 ng/mL for OA and 5.19 ng/mL for STX. In summary, all of the results indicate that the dual functional cardiomyocyte-based hybrid-biosensor will be a promising and utility tool for shellfish toxin detection.


Assuntos
Técnicas Biossensoriais , Miócitos Cardíacos/metabolismo , Ácido Okadáico/análise , Saxitoxina/análise , Intoxicação por Frutos do Mar/diagnóstico , Animais , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Miócitos Cardíacos/citologia , Ácido Okadáico/farmacologia , Ratos , Ratos Sprague-Dawley , Saxitoxina/farmacologia , Frutos do Mar
17.
J Environ Radioact ; 192: 485-490, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30103169

RESUMO

Paralytic shellfish poisoning (PSP) events occur regularly along the Mediterranean and Atlantic coast of Morocco, and have been responsible for several severe cases of human intoxication. Along the southern Atlantic coast of Morocco, aquaculture and intensive artisanal fishing practices have recently been particularly heavily impacted, and toxic species have been observed in increasing intensity and frequency. In the 1990's a regulatory monitoring program was established for the coastal waters off Morocco by the National Institute of Fisheries Research (INRH), to reduce the risk of intoxication with biotoxins. The regulatory monitoring is conducted weekly and includes toxic phytoplankton enumeration and identification, as well as saxitoxin (STX) analysis in seafood using the mouse bioassay (MBA). Animal testing remains the most widely used screening method for PSP toxin detection, yet its use is being reconsidered for animal-related ethical issues, as well as for practical considerations. To be able to better evaluate alternatives to animal testing, the performance of a nuclear-based radioligand-receptor binding assay (RBA) for paralytic shellfish toxins was assessed and compared with the MBA using four commercially important shellfish matrices, including cockles Cerastoderma edule, razor shells Solen marginatus, oysters Crassostrea gigas, and mussels Perna perna. Over 50 samples were collected and analysed as part of the regulatory monitoring framework including a suite of monthly samples from 2017 and all samples identified as toxic by MBA since 2011. Testing of reference material and evaluation of assay-critical parameters (e.g. slope of calibration curve, internal quality control QC and IC50) confirmed the robustness of the RBA methodology. With this RBA method, STX-like activity detected in shellfish samples ranged from 33 to 8500 µg STX equivalents per kg. RBA data were significantly correlated (P < 0.0001, Pearson r = 0.96) with the MBA-derived dataset. Importantly, the RBA method allowed for the detection and quantification of PSP toxins at levels not detectable by using the mouse bioassay. The limits of quantification of the RBA was calculated and found to be 10-fold lower than that of the MBA, respectively 35.24 ±â€¯5.99 and 325 µg STX equivalents per kg of tissue. In addition, the RBA was easier to use and produced reliable results more rapidly than the MBA and without use of live animals. Considering the increasing risks associated with harmful algal blooms, globally and in Morocco, together with the increased development of aquaculture production and seafood consumption and the difficulties of live animal testing, these findings indicate that the RBA method is a reliable and effective alternative to the MBA method.


Assuntos
Contaminação de Alimentos/análise , Ensaio Radioligante/métodos , Saxitoxina/análise , Animais , Humanos , Camundongos , Marrocos , Perna (Organismo)
18.
ACS Sens ; 3(7): 1233-1245, 2018 07 27.
Artigo em Inglês | MEDLINE | ID: mdl-29974739

RESUMO

Due to increasing stringency of water legislation and extreme consequences that failure to detect some contaminants in water can involve, there has been a strong interest in developing electrochemical biosensors for algal toxin detection during the past decade, evidenced by literature increasing from 2 journal papers pre-2009 to 24 between 2009 and 2018. In this context, this review has summarized recent progress of successful algal toxin detection in water using electrochemical biosensing techniques. Satisfactory detection recoveries using real environmental water samples and good sensor repeatability and reproducibility have been achieved, along with some excellent limit-of-detection (LOD) reported. Recent electrochemical biosensor literature in algal toxin detection is compared and discussed to cover three major design components: (1) biorecognition elements, (2) electrochemical read-out techniques, and (3) sensor electrodes and signal amplification strategy. The recent development of electrochemical biosensors has provided one more step further toward quick in situ detection of algal toxins in the contamination point of the water source. In the end, we have also critically reviewed the current challenges and research opportunities regarding electrochemical biosensors for algal toxin detection that need to be addressed before they attain commercial viability.


Assuntos
Toxinas Bacterianas/análise , Técnicas Biossensoriais/instrumentação , Cianobactérias/química , Dinoflagelados/química , Técnicas Eletroquímicas/instrumentação , Toxinas Marinhas/análise , Microcistinas/análise , Poluentes Químicos da Água/análise , Água/análise , Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/métodos , Monitoramento Ambiental/instrumentação , Monitoramento Ambiental/métodos , Desenho de Equipamento , Neurotoxinas/análise , Saxitoxina/análise
19.
Spectrochim Acta A Mol Biomol Spectrosc ; 204: 180-187, 2018 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-29933153

RESUMO

Aptamers could be used to construct simple and effective biosensor because the conformational switch of aptamer upon target binding is easy to be transferred to optical or electrochemical signals. Nevertheless, we found that the binding between saxitoxin (STX) and aptamer (M-30f) is not accompanied with conformational switch. Here, the circular dichroism spectra, fluorophore and quencher labeled aptamer, and crystal violet-based assays were used to identify the binding way between STX and aptamer. The results show that the conformation of aptamer is stabilized in PBS buffer (10 mM phosphate buffer, 2.7 mM KCl, 137 mM NaCl, pH 7.4) and this conformation may provide an exactly suitable cave for STX binding. Through the analysis of UV-melting curves and circular dichroism-melting curves, it is found that different concentrations of STX produce different unfolding extents of the aptamer under high temperature. Then, a simple temperature-assisted "turn-on" fluorescent aptasensor was developed to detect STX and the application in real sample detection demonstrates its feasibility. The proposed method provides not only an alternative for STX detection but also a strategy for simple aptasensor design using aptamers that do not switch conformation upon targets binding.


Assuntos
Aptâmeros de Nucleotídeos/metabolismo , Técnicas Biossensoriais/métodos , Corantes Fluorescentes/metabolismo , Saxitoxina/análise , Saxitoxina/metabolismo , Aptâmeros de Nucleotídeos/química , Corantes Fluorescentes/química , Limite de Detecção , Modelos Lineares , Reprodutibilidade dos Testes , Saxitoxina/química
20.
Food Res Int ; 108: 274-279, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29735058

RESUMO

Paralytic shellfish poisoning is caused by saxitoxin and its analogues. The paralytic shellfish toxins (PSTs) are produced by marine dinoflagellates and can be accumulated in filter feeding shellfish, such as mussel, clam, oyster and ark shell. The worldwide regulatory limits for PSTs in shellfish are set at 80 µg STX eq./100 g meat and this is widely accepted as providing adequate public health protection. In this study, we have determined five individual PSTs (STX, GTX1, GTX2, GTX3 and GTX4) in shellfish using LC-MS/MS and assessed the human acute and chronic exposures to PSTs through shellfish consumption. Food consumption data was obtained from the Korea National Health and Nutrition Examination Survey (KNHANES 2010-2015). The acute exposure using a large portion size of 88 g/day (95th percentile for consumers only) with maximum toxin level of 198.7 µg/kg was 0.30 µg/kg bw. Even though we estimated the acute exposure with a conservative manner, it was below the ARfDs (0.5 or 0.7 µg STX eq./kg bw) proposed by the international organizations, representing 43-60% of the ARfDs. The chronic exposures using mean consumption data for whole population with mean concentration of PSTs were ranged from 0.002 to 0.026 µg STX eq./kg bw/day. For consumers only, the chronic exposures were in the range of 0.012-0.128 µg STX eq./kg bw/day.


Assuntos
Toxinas Bacterianas/análise , Exposição Dietética , Análise de Alimentos/métodos , Saxitoxina/análise , Alimentos Marinhos/análise , Intoxicação por Frutos do Mar/etiologia , Frutos do Mar/análise , Adolescente , Adulto , Idoso , Toxinas Bacterianas/efeitos adversos , Criança , Pré-Escolar , Cromatografia Líquida , Exposição Dietética/efeitos adversos , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Inquéritos Nutricionais , República da Coreia , Medição de Risco , Fatores de Risco , Saxitoxina/efeitos adversos , Saxitoxina/análogos & derivados , Alimentos Marinhos/efeitos adversos , Frutos do Mar/efeitos adversos , Intoxicação por Frutos do Mar/diagnóstico , Espectrometria de Massas em Tandem , Fatores de Tempo , Adulto Jovem
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