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1.
Food Chem ; 368: 130856, 2022 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-34425333

RESUMO

In this work, we present a novel dual-emissive fluoroimmunoassay for synchronous monitoring of okadaic acid (OA) and saxitoxin (STX) using multicolor fluorescent labels composed of sulfur, phosphorous co-doped graphene quantum dots (S, P-GQDs), and ovalbumin (OVA)-coated gold nanoparticles (OVA-AuNPs). The novel OVA-AuNPs were prepared by the reduction of chloroauric acid under alkaline conditions using OVA as a reducing agent. Both S, P-GQDs and OVA-AuNPs exhibit bright fluorescence, more importantly, a large emission wavelength difference (Δλ = 156 nm) under an excitation of 400 nm and relatively independent fluorescence behavior, which are essential to realizing the dual-signal marks in a directly mixing system. Using a competitive fluorescence-linked immunosorbent assay (cFLISA) format, the dual-emissive cFLISA was successfully utilized to measure OA and STX contents in Alectryonella plicatula (commonly named as fingerprint oyster) and the detection results were in good agreement with the commercial enzyme-linked immunosorbent assay (ELISA) kits.


Assuntos
Grafite , Nanopartículas Metálicas , Pontos Quânticos , Ouro , Imunoensaio , Ácido Okadáico , Saxitoxina , Frutos do Mar/análise
2.
Sci Total Environ ; 802: 149801, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34454155

RESUMO

Temperature may affect the production of saxitoxin (STX) and its derivatives (STXs); however, this is still controversial. Further, STX-biosynthesis gene regulation and the relation of its toxicity with temperature are not clearly understood. In the present study, we evaluated the effects of different temperatures (12 °C, 16 °C, and 20 °C) on the growth, toxin profiles, and expression of two core STX-biosynthesis genes, sxtA and sxtG, in the toxic dinoflagellate Alexandrium pacificum Alex05, isolated from Korean coasts. We found that temperature significantly affected cell growth, with maximum growth recorded at 16 °C, followed by 20 °C and 12 °C. HPLC analysis revealed mostly 12 of STXs from the tested cultures. Interestingly, the contents of STXs increased in the cells cultured at 16 °C and exposed to cold stress, compared to the 20 °C culture and heat stress; however, toxin components were much more diverse under heat stress. These toxin profiles generally matched with the sxtA and sxtG expression levels. Incubation at lower temperatures (12 °C and 16 °C) and exposure to cold stress increased sxtA and sxtG expressions in the cells, whereas heat stress showed little change or downregulated the transcription of both genes. Principal component analysis (PCA) showed low correlation between STXs eq and expressional levels of sxtA and sxtG in heat-stressed cells. These results suggest that temperature might be a crucial factor affecting the level and biosynthesis of STXs in marine toxic dinoflagellates.


Assuntos
Dinoflagelados , Saxitoxina , Dinoflagelados/genética , Expressão Gênica , Regulação da Expressão Gênica , Saxitoxina/toxicidade , Temperatura
3.
Toxicon ; 204: 56-63, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34742781

RESUMO

Saxitoxin and its derivatives, the paralytic shellfish toxins (PSTs), are well known to be toxic to humans, and maximum permitted levels in seafood have been established by regulatory authorities in many countries. Monitoring of PSTs is typically performed using chemical methods which quantify the concentration of the individual PST analogues, of which there are many. However, since the toxicities of analogues are different, they do not equally contribute to the overall toxicity of the sample. To account for these differences, toxicity equivalency factors (TEFs) need to be determined for each analogue and applied. Currently there are no established TEFs for decarbamoyl gonyautoxin 1&4 (dcGTX1&4), which occurs in some clam species such as Mactra chinensis contaminated with PSTs due to metabolism within the shellfish. In this study the median lethal dose of purified, equilibrated epimeric mixture of dcGTX1&4 has been determined by intraperitoneal injection (i.p.) (4.75 µmol/kg) and by feeding (34.9 µmol/kg). The most relevant route of exposure is orally with feeding being more representative of human consumption and more reliable than gavage. Based on the median lethal dose by feeding, a TEF of 0.1 is recommended for dcGTX1&4. Receptor binding activity and i.p. toxicity results showed dcGTX1&4 to be much less toxic than STX (140-170-fold). However, by feeding a much smaller difference in toxicity was observed with dcGTX1&4 being only 11-fold less toxic than STX. Analysis of the gut contents of mice dosed with dcGTX1&4 showed the presence of decarbamoyl gonyautoxin 2&3, decarbamoyl saxitoxin and decarbamoyl neosaxitoxin, all of which are of greater toxicity. This conversion of dcGTX1&4 within the digestive track to more toxic congeners may explain the high relative toxicity of dcGTX1&4 by feeding compared to that determined by i.p. and by sodium channel activity.


Assuntos
Bivalves , Intoxicação por Frutos do Mar , Animais , Camundongos , Saxitoxina/análogos & derivados , Saxitoxina/toxicidade , Frutos do Mar/análise
4.
Rev Environ Contam Toxicol ; 258: 109-150, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34622370

RESUMO

Harmful cyanobacterial blooms are increasing and becoming a worldwide concern as many bloom-forming cyanobacterial species can produce toxic metabolites named cyanotoxins. These include microcystins, saxitoxins, anatoxins, nodularins, and cylindrospermopsins, which can adversely affect humans, animals, and the environment. Different methods to assess these classes of compounds in vitro and in vivo include biological, biochemical, molecular, and physicochemical techniques. Furthermore, toxic effects not attributable to known cyanotoxins can be observed when assessing bloom material. In order to determine exposures to cyanotoxins and to monitor compliance with drinking and bathing water guidelines, it is necessary to have reliable and effective methods for the analysis of these compounds. Many relatively simple low-cost methods can be employed to rapidly evaluate the potential hazard. The main objective of this mini-review is to describe the assessment of toxic cyanobacterial samples using in vitro and in vivo bioassays. Newly emerging cyanotoxins, the toxicity of analogs, or the interaction of cyanobacteria and cyanotoxins with other toxicants, among others, still requires bioassay assessment. This review focuses on some biological and biochemical assays (MTT assay, Immunohistochemistry, Micronucleus Assay, Artemia salina assay, Daphnia magna test, Radionuclide recovery, Neutral red cytotoxicity and Comet assay, Enzyme-Linked Immunosorbent Assay (ELISA), Annexin V-FITC assay and Protein Phosphatase Inhibition Assay (PPIA)) for the detection and measurement of cyanotoxins including microcystins, cylindrospermopsins, anatoxin-a, saxitoxins, and nodularins. Although most bioassay analyses often confirm the presence of cyanotoxins at low concentrations, such bioassays can be used to determine whether some strains or blooms of cyanobacteria may produce other, as yet unknown toxic metabolites. This review also aims to identify research needs and data gaps concerning the toxicity assessment of cyanobacteria.


Assuntos
Cianobactérias , Microcistinas , Animais , Humanos , Microcistinas/toxicidade , Saxitoxina , Uracila
5.
Toxicon ; 204: 5-8, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34666135

RESUMO

The Osteoarthritis is a chronic disease characterized by a progressive deterioration of the articular cartilage producing a strong inflammatory activity and chronic pain in patients. Horses also show osteoarthritis. Since the activation and progression of the disease are similar to that of human we developed a study model in horses. In this study, we test the effect of Neosaxitoxin, a phycotoxin from Paralytic Shellfish Poison, in the remediation of osteoarthritis equine clinical symptoms such as pain (showed in lameness) and inflammation quantifying the amounts of pro-inflammatory markers like cellular infiltration, TNF-alpha and nitric oxide in the synovial fluid obtained from the horse damaged joint. The outcomes show that Neosaxitoxin blocks pain for long lasting period (average 24.7 days). Furthermore, the amounts of pro-inflammatory markers were reduced and consequently an enhanced horse's well-being was obtained. Neosaxitoxin showed to be a candidate for establishing treatment protocols for OA, being safe and effective as a pain blocker in equine osteoarthritis.


Assuntos
Doenças dos Cavalos , Osteoartrite , Venenos , Animais , Doenças dos Cavalos/induzido quimicamente , Doenças dos Cavalos/tratamento farmacológico , Cavalos , Humanos , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Inflamação/veterinária , Osteoartrite/induzido quimicamente , Osteoartrite/tratamento farmacológico , Osteoartrite/veterinária , Dor/tratamento farmacológico , Dor/veterinária , Saxitoxina/análogos & derivados , Frutos do Mar
6.
Harmful Algae ; 105: 102068, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-34303514

RESUMO

Harmful algal bloom events are increasing in a number of water bodies around the world with significant economic impacts on the aquaculture, fishing and tourism industries. As well as their potential impacts on human health, toxin exposure from harmful algal blooms (HABs) has resulted in widespread morbidity and mortality in marine life, including top marine predators. There is therefore a need for an improved understanding of the trophic transfer, and persistence of toxins in marine food webs. For the first time, the concentrations of two toxin groups of commercial and environmental importance, domoic acid (DA) and saxitoxin (including Paralytic Shellfish Toxin (PST) analogues), were measured in the viscera of 40 different fish species caught in Scotland between February and November, 2012 to 2019. Overall, fish had higher concentrations of DA compared to PSTs, with a peak in the summer / autumn months. Whole fish concentrations were highest in pelagic species including Atlantic mackerel and herring, key forage fish for marine predators including seals, cetaceans and seabirds. The highest DA concentrations were measured along the east coast of Scotland and in Orkney. PSTs showed highest concentrations in early summer, consistent with phytoplankton bloom timings. The detection of multiple toxins in such a range of demersal, pelagic and benthic fish prey species suggests that both the fish, and by extension, piscivorous marine predators, experience multiple routes of toxin exposure. Risk assessment models to understand the impacts of exposure to HAB toxins on marine predators therefore need to consider how chronic, low-dose exposure to multiple toxins, as well as acute exposure during a bloom, could lead to potential long-term health effects ultimately contributing to mortalities. The potential synergistic, neurotoxic and physiological effects of long-term exposure to multiple toxins require investigation in order to appropriately assess the risks of HAB toxins to fish as well as their predators.


Assuntos
Proliferação Nociva de Algas , Saxitoxina , Animais , Cadeia Alimentar , Humanos , Fitoplâncton , Escócia
7.
Harmful Algae ; 105: 102056, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-34303515

RESUMO

Alexandrium catenella is a harmful algal bloom (HAB)-forming dinoflagellate that causes significant damage to the cultivation and harvest of shellfish due to its synthesis of paralytic shellfish toxins.  To evaluate the potential for macroalgae aquaculture to mitigate A. catenella blooms, we determined the effects of three cultivable macroalgae - Saccharina latissima (sugar kelp), Chondrus crispus (Irish moss), and Ulva spp. - on A. catenella in culture- and field-based experiments.  Co-culture growth assays of A. catenella exposed to environmentally realistic concentrations of each macroalgae showed that all species except low levels of C. crispus caused cell lysis and significant reductions in A. catenella densities relative to control treatments of 17-74% in 2-3 days and 42-96% in ~one week (p<0.05 for all assays). In a toxin accumulation experiment, S. latissima significantly lessened (p<0.05) saxitoxin (STX) accumulation in blue mussels (Mytilus edulis), keeping levels (71.80±1.98 µg STX 100 g-1) below US closure limits (80 µg STX 100 g-1) compared to the untreated control (93.47±8.11 µg STX 100 g-1). Bottle incubations of field-collected, bloom populations of A. catenella experienced significant reductions in cell densities of up to 95% when exposed to aquaculture concentrations of all three macroalgae (p<0.005 for all). The stocking of aquacultured S. latissima within mesocosms containing a bloom population of A. catenella (initial density: 3.2 × 104 cells L-1) reduced the population of A. catenella by 73% over 48 h (p<0.005) while Ulva addition caused a 54% reduction in A. catenella over 96 h (p<0.01).  Among the three seaweeds, their ordered ability to inhibit A. catenella was S. latissima > Ulva spp. > C. crispus. Seaweeds' primary anti-A. catenella activity were allelopathic, while nutrient competition, pH elevation, and macroalgae-attached bacteria may have played a contributory role in some experiments. Collectively, these results suggest that the integration of macroalgae with shellfish-centric aquaculture establishments should be considered as a non-invasive, environmentally friendly, and potentially profit-generating measure to mitigate A. catenella-caused damage to the shellfish aquaculture industry.


Assuntos
Bivalves , Dinoflagelados , Alga Marinha , Animais , Proliferação Nociva de Algas , Saxitoxina
8.
Sensors (Basel) ; 21(14)2021 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-34300682

RESUMO

Saxitoxin (STX) belongs to the family of marine biological toxins, which are major contaminants in seafood. The reference methods for STX detection are mouse bioassay and chromatographic analysis, which are time-consuming, high costs, and requirement of sophisticated operation. Therefore, the development of alternative methods for STX analysis is urgent. Electrochemical analysis is a fast, low-cost, and sensitive method for biomolecules analysis. Thus, in this study, an electrolyte-insulator-semiconductor (EIS) sensor based on aptamer-modified two-dimensional layered Ti3C2Tx nanosheets was developed for STX detection. The high surface area and rich functional groups of MXene benefited the modification of aptamer, which had specific interactions with STX. Capacitance-voltage (C-V) and constant-capacitance (ConCap) measurement results indicated that the aptasensor was able to detect STX with high sensitivity and good specificity. The detection range was 1.0 nM to 200 nM and detection limit was as low as 0.03 nM. Moreover, the aptasensor was found to have a good selectivity and two-week stability. The mussel tissue extraction test suggested the potential application of this biosensor in detecting STX in real samples. This method provides a convenient approach for low-cost, rapid, and label-free detection of marine biological toxins.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Animais , Técnicas Eletroquímicas , Limite de Detecção , Toxinas Marinhas , Camundongos , Saxitoxina , Titânio
9.
Aging (Albany NY) ; 13(13): 17211-17226, 2021 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-34197336

RESUMO

Saxitoxin (STX), as a type of paralytic shellfish poisoning (PSP), is gaining widespread attention due to its long existence in edible shellfish. However, the mechanism underlying STX chronic exposure-induced effect is not well understood. Here, we evaluated the neurotoxicity effects of long-term low-dose STX exposure on C57/BL mice by behavioral tests, pathology analysis, and hippocampal proteomics analysis. Several behavioral tests showed that mice were in a cognitive deficiency after treated with 0, 0.5, 1.5, or 4.5 µg STX equivalents/kg body weight in the drinking water for 3 months. Compared with control mice, STX-exposed mice exhibited brain neuronal damage characterized by decreasing neuronal cells and thinner pyramidal cell layers in the hippocampal CA1 region. A total of 29 proteins were significantly altered in different STX dose groups. Bioinformatics analysis showed that protein phosphatase 1 (Ppp1c) and arylsulfatase A (Arsa) were involved in the hippo signaling pathway and sphingolipid metabolism pathway. The decreased expression of Arsa indicates that long-term low doses of STX exposure can cause neuronal inhibition, which is a process related to spatial memory impairment. Taken together, our study provides a new understanding of the molecular mechanisms of STX neurotoxicity.


Assuntos
Síndromes Neurotóxicas/patologia , Saxitoxina/toxicidade , Animais , Comportamento Animal/efeitos dos fármacos , Região CA1 Hipocampal/patologia , Transtornos Cognitivos/induzido quimicamente , Transtornos Cognitivos/psicologia , Relação Dose-Resposta a Droga , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Transtornos da Memória/induzido quimicamente , Transtornos da Memória/psicologia , Camundongos , Camundongos Endogâmicos C57BL , Neurônios/patologia , Síndromes Neurotóxicas/psicologia , Proteômica , Células Piramidais/patologia
10.
Nat Commun ; 12(1): 4171, 2021 07 07.
Artigo em Inglês | MEDLINE | ID: mdl-34234116

RESUMO

Here we report the pharmacologic blockade of voltage-gated sodium ion channels (NaVs) by a synthetic saxitoxin derivative affixed to a photocleavable protecting group. We demonstrate that a functionalized saxitoxin (STX-eac) enables exquisite spatiotemporal control of NaVs to interrupt action potentials in dissociated neurons and nerve fiber bundles. The photo-uncaged inhibitor (STX-ea) is a nanomolar potent, reversible binder of NaVs. We use STX-eac to reveal differential susceptibility of myelinated and unmyelinated axons in the corpus callosum to NaV-dependent alterations in action potential propagation, with unmyelinated axons preferentially showing reduced action potential fidelity under conditions of partial NaV block. These results validate STX-eac as a high precision tool for robust photocontrol of neuronal excitability and action potential generation.


Assuntos
Potenciais de Ação/efeitos dos fármacos , Canal de Sódio Disparado por Voltagem NAV1.2/metabolismo , Saxitoxina/farmacologia , Bloqueadores do Canal de Sódio Disparado por Voltagem/farmacologia , Animais , Axônios/efeitos dos fármacos , Axônios/metabolismo , Células CHO , Células Cultivadas , Corpo Caloso/citologia , Corpo Caloso/efeitos dos fármacos , Corpo Caloso/metabolismo , Cricetulus , Embrião de Mamíferos , Feminino , Hipocampo/citologia , Masculino , Camundongos , Canal de Sódio Disparado por Voltagem NAV1.2/genética , Técnicas de Patch-Clamp , Cultura Primária de Células , Ratos , Ratos Sprague-Dawley , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Saxitoxina/análogos & derivados , Saxitoxina/efeitos da radiação , Análise de Célula Única , Análise Espaço-Temporal , Raios Ultravioleta , Bloqueadores do Canal de Sódio Disparado por Voltagem/efeitos da radiação
11.
Shokuhin Eiseigaku Zasshi ; 62(3): 85-93, 2021.
Artigo em Japonês | MEDLINE | ID: mdl-34219101

RESUMO

The mouse bioassay (MBA) for paralytic shellfish toxins (PSTs) in bivalves has been used as an official method in Japan. It is necessary to develop an alternative method to animal experiments in PSTs assay because 3Rs (Replacement, Reduction, and Refinement) of animal experiments are required from the animal welfare point of view. Various methods such as HPLC-FL, receptor binding assay, LC-MS/MS and ELISA have been established to detect PSTs without performing animal experiments. The present study was undertaken to develop a screening method using oligonucleotide lateral flow immunoassay (OLFIA) for detecting PSTs in bivalves. The screening level was defined as positive at 2 MU/g of MBA that is the half regulation limit of PSTs monitoring in Japan. All 20 positive (equal to or more than 2 MU/g) samples judged from MBA showed a positive reaction in the OLFIA. No positive samples resulted in a false negative reaction. The OLFIA exhibited high accuracy at 2 MU/g of screening criteria. The authors demonstrated here that the OLFIA can be useful for rapid detection of PSTs in bivalves.


Assuntos
Bivalves , Intoxicação por Frutos do Mar , Animais , Cromatografia Líquida , Imunoensaio , Japão , Toxinas Marinhas/análise , Camundongos , Oligonucleotídeos , Saxitoxina/análise , Frutos do Mar/análise , Espectrometria de Massas em Tandem
12.
J Chromatogr A ; 1651: 462328, 2021 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-34153733

RESUMO

Paralytic shellfish toxins and tetrodotoxin (puffer-fish toxin), the latter of which was recently found in bivalves from Europe, Japan, and New Zealand, are potent neurotoxins. A simple and effective clean-up procedure was developed for the simultaneous determination of ten paralytic shellfish toxins (gonyautoxins 1-6, decarbamoylgonyautoxins 2 and 3, and N-sulfocarbamoylgonyautoxins 2 and 3) and tetrodotoxin in the scallop, Mizuhopecten (Patinopecten) yessoensis, and the short-necked clam, Ruditapes philippinarum. To reduce matrix effects, 1% aqueous acetic acid extracts of the bivalves were cleaned up by ion-pair solid-phase extraction using a graphite carbon cartridge with tridecafluoroheptanoic acid as the volatile ion-pair reagent, followed by fourfold dilution. The ten paralytic shellfish toxins and tetrodotoxin were then separated on a hydrophilic interaction chromatography column and quantified by tandem mass spectrometry. The limits of detection and the limits of quantification for the ten PSTs ranged from 0.09 to 13.0 µg saxitoxin equivalents/kg and from 0.26 to 39.4 µg saxitoxin equivalents/kg, respectively. The limit of detection and the limit of quantification for tetrodotoxin ranged from 27.4 to 27.9 µg/kg and from 83.1 to 84.4 µg/kg, respectively. The proposed method yielded minimal matrix effects for the 11 analytes, thus allowing their quantification by simple external calibration. The proposed method also gave good mean recoveries of the 11 analytes ranging from 75.7 to 96.2% with relative standard deviations less than 16% at three fortification levels for the ten paralytic shellfish toxins (total concentrations of 277, 554, and 1107 µg saxitoxin equivalents/kg) and tetrodotoxin (100, 200, and 400 µg/kg) in the two bivalve samples. Finally, the proposed method was applied for the determination of the ten paralytic shellfish toxins and tetrodotoxin in scallop and short-necked clam samples.


Assuntos
Bivalves/química , Pectinidae/química , Saxitoxina/análogos & derivados , Extração em Fase Sólida , Espectrometria de Massas em Tandem , Tetrodotoxina/análise , Animais , Europa (Continente) , Grafite/química , Interações Hidrofóbicas e Hidrofílicas , Saxitoxina/análise , Alimentos Marinhos/análise , Tetrodotoxina/isolamento & purificação
13.
Anal Chim Acta ; 1173: 338710, 2021 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-34172145

RESUMO

Saxitoxin (STX) is a small molecule toxin (Mw. ca. 299 g/mol) with high acute toxicity, and it has urgent need of facile analytical methods. Herein, a competitive colorimetric aptasensor was developed for highly sensitive detection of STX. An anti-STX aptamer was hybridized with a complementary strand on the magnetic beads and was competitively bound by STX. The supernatant containing the aptamer binding to STX was obtained by magnetic separation, which could trigger hybridization chain reaction (HCR) to generate rigid double stranded DNAs (dsDNAs) with sticky end and variable length. These HCR-dsDNAs were found to be able to facilitate significant enhancement on the peroxidase-like catalytic capability of AuNPs nanozyme towards 3,3,5,5-tetramethylbenzidine (TMB). The concentration of STX was responded in a "turn on" mode, based on the amplified colorimetric transduction thereof. The aptasensor realized high sensitivity, with a limit of detection (LOD) as low as 42.46 pM. Moreover, a wide linear detection range of 78.13-2500 pM, good selectivity, as well as good recovery rates of 106.2-113.5% when analyzing STX in real shellfish samples were obtained. This strategy could be referred to develop robust aptasensors for simple and highly sensitive detection of other small molecules and toxins.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Nanopartículas Metálicas , Catálise , Colorimetria , Ouro , Limite de Detecção , Saxitoxina
14.
Mar Drugs ; 19(6)2021 May 21.
Artigo em Inglês | MEDLINE | ID: mdl-34064031

RESUMO

Toxic dinoflagellate Alexandrium spp. produce saxitoxins (STXs), whose biosynthesis pathway is affected by temperature. However, the link between the regulation of the relevant genes and STXs' accumulation and temperature is insufficiently understood. In the present study, we evaluated the effects of temperature on cellular STXs and the expression of two core STX biosynthesis genes (sxtA4 and sxtG) in the toxic dinoflagellate Alexandrium catenella Alex03 isolated from Korean waters. We analyzed the growth rate, toxin profiles, and gene responses in cells exposed to different temperatures, including long-term adaptation (12, 16, and 20 °C) and cold and heat stresses. Temperature significantly affected the growth of A. catenella, with optimal growth (0.49 division/day) at 16 °C and the largest cell size (30.5 µm) at 12 °C. High concentration of STXs eq were detected in cells cultured at 16 °C (86.3 fmol/cell) and exposed to cold stress at 20→12 °C (96.6 fmol/cell) compared to those at 20 °C and exposed to heat stress. Quantitative real-time PCR (qRT-PCR) revealed significant gene expression changes of sxtA4 in cells cultured at 16 °C (1.8-fold) and cold shock at 20→16 °C (9.9-fold). In addition, sxtG was significantly induced in cells exposed to cold shocks (20→16 °C; 19.5-fold) and heat stress (12→20 °C; 25.6-fold). Principal component analysis (PCA) revealed that low temperature (12 and 16 °C) and cold stress were positively related with STXs' production and gene expression levels. These results suggest that temperature may affect the toxicity and regulation of STX biosynthesis genes in dinoflagellates.


Assuntos
Dinoflagelados/genética , Dinoflagelados/metabolismo , Biossíntese de Proteínas/genética , Proteínas de Protozoários/metabolismo , Saxitoxina/biossíntese , Saxitoxina/genética , Crescimento Celular , Proliferação de Células , Temperatura Baixa , Resposta ao Choque Frio , Dinoflagelados/crescimento & desenvolvimento , Regulação da Expressão Gênica , Análise de Componente Principal , Proteínas de Protozoários/genética
15.
Harmful Algae ; 103: 101981, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33980431

RESUMO

As harmful algal blooms (HABs) increase in magnitude and duration worldwide, they are becoming an expanding threat to marine wildlife. Over the past decade, blooms of algae that produce the neurotoxins domoic acid (DA) and saxitoxin (STX) and documented concurrent seabird mortality events have increased bicoastally in the United States. We conducted a retrospective analysis of HAB related mortality events in California, Washington, and Rhode Island between 2007 and 2018 involving 12 species of seabirds, to document the levels, ranges, and patterns of DA and STX in eight sample types (kidney, liver, stomach, intestinal, cloacal, cecal contents, bile, blood) collected from birds during these events. Samples (n = 182) from 83 birds were examined for DA (n = 135) or STX (n = 17) or both toxins simultaneously (n = 30), using ELISA or LCMS at the National Oceanographic and Atmospheric Administration, National Marine Fisheries Service (NOAA-NMFS) Wildlife Algal-toxin Research and Response Network (WARRN-West) or the University of California, Santa Cruz (UCSC). DA or STX was detected in seven of the sample types with STX below the minimum detection limit in blood for the three samples tested. DA was found in 70% and STX was found in 23% of all tested samples. The ranges of detectable levels of DA and STX in all samples were 0.65-681,190.00 ng g-1 and 2.00-20.95 ng g-1, respectively. Cloacal contents from a Pacific loon (Gavia pacifica) collected in 2017 from Ventura County, California, had the highest maximum level of DA for all samples and species tested in this study. The highest level of STX for all samples and species was detected in the bile of a northern fulmar (Fulmarus glacialis) collected in 2018 from San Luis Obispo County, California. DA detections were consistently found in gastrointestinal samples, liver, bile, and kidney, whereas STX detections were most frequently seen in liver and bile samples. Co-occurring HAB toxins (DA and STX) were detected in white-winged scoters (Melanitta deglandi) in 2009, a Brandt's cormorant (Phalacrocorax penicillatus) in 2015, and a northern fulmar and common murre (Uria aalge) in 2018. This article provides DA and STX tissue concentrations and patterns in avian samples and shows the utility of various sample types for the detection of HAB toxins. Future research to understand the pharmacodynamics of these toxins in avian species and to establish lethal doses in various bird species would be beneficial.


Assuntos
Aves , Saxitoxina , Animais , Ácido Caínico/análogos & derivados , Estudos Retrospectivos , Rhode Island , Estados Unidos , Washington
16.
Harmful Algae ; 103: 102000, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33980440

RESUMO

Published baseline data on biotoxin exposure in cetaceans is sparse but critical for interpreting mortality events as harmful algal blooms increase in frequency and duration. We present the first synthesis of domoic acid (DA), saxitoxin (STX), okadaic acid (OA), and microcystin detections in the feces and urine of stranded and bycaught southern California cetaceans, over an 18 year period (2001-2018), along with corresponding stomach content data. DA was detected in 13 out of 19 cetacean species, most often in harbor porpoise (Phocoena phocoena) (81.8%, n = 22) and long-beaked common dolphins (Delphinus delphis bairdii) (74%, n = 231). Maximum DA concentrations of 324,000 ng/g in feces and 271, 967 ng/ml in urine were observed in D. d. bairdii. DA was detected more frequently and at higher concentrations in male vs. female D. d. bairdii. Higher fecal DA concentrations in D. d. bairdii were associated with a greater proportion of northern anchovy (Engraulis mordax) in the diet, indicating it may be a primary vector of DA. Fecal DA concentrations for D. d. bairdii off Point Conception were greater than those from animals sampled off Los Angeles and San Diego counties, reflecting greater primary productivity and higher Pseudo-nitzschia spp. abundance in that region and a greater abundance of E. mordax in the diet. STX was detected at low levels (fecal max = 7.5 ng/g, urine max = 17 ng/ml) in 3.6% (n = 165) of individuals from 3 out of 11 species. The occurrence of E. mordax in 100% of the 3 examined stomachs suggests this species could be a primary vector of the detected STX. OA was detected in 2.4% of tested individuals (n = 85) at a maximum fecal concentration of 422.8 ng/g. Microcystin was detected in 14.3% (n = 7) of tested individuals with a maximum liver concentration of 96.8 ppb.


Assuntos
Monitoramento Ambiental , Saxitoxina , Animais , California , Cetáceos , Proliferação Nociva de Algas , Saxitoxina/análise
17.
Harmful Algae ; 103: 102004, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33980444

RESUMO

Raphidiopsis raciborskii (formerly Cylindrospermopsis raciborskii) is a freshwater cyanobacterium potentially producing saxitoxins (STX) and cylindrospermopsin. Its ecophysiological versatility enables it to form blooms in the most diverse types of environments, from tropical to temperate, and from relatively pristine to polluted. In Peri Lake, located in the subtropical south of Brazil, growing populations of STX-producing R. raciborskii have been detected since 1994, posing risks to the use of its waters that supply a population of about 100,000 inhabitants. Despite the existence of a monitoring system for the presence and toxicity of cyanobacteria in Peri Lake water, no assessment has been made in the coastal region, downstream of outflowing lake water, thereby potentially making available a toxic biomass to natural and cultivated shellfish populations in the salt water ecosystem. To address this problem, the present study evaluated environmental variables and STX concentration by profiling the outflowing waters between Peri Lake and the adjacent coastal zone. Laboratory experiments were carried out with three strains of R. raciborskii in order to confirm the effect of salinity on STX production and verify if Perna Perna mussels fed with R. raciborskii cultures would absorb and accumulate STX. Results showed that environmental concentrations of STX reach high levels (up to 6.31 µg L-1 STX eq.), especially in the warmer months, reaching the coastal zone. In laboratory tests, it was found that the strains tolerate salinities between 4 and 6 and that salinity influences the production of STX. In addition, mussels fed with R. raciborskii effectively absorb and accumulate STX, even in typically marine salinities (22 to 30), suggesting that R. raciborskii biomass remains available and toxic despite salinity shock. These results draw attention to the ecological and health risk associated with R. raciborskii blooms, both in the lake environment and in the adjacent marine environment, calling attention to the need to improve the monitoring and management systems for water and shellfish toxicity in the region of interest, as well as other places where toxic cyanobacteria of limnic origin can reach the coastal zone.


Assuntos
Bivalves , Cianobactérias , Animais , Brasil , Cylindrospermopsis , Ecossistema , Saxitoxina
18.
Harmful Algae ; 103: 102012, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33980451

RESUMO

Harmful algal blooms that can produce toxins are common in the Indian River Lagoon (IRL), which covers ~250 km of Florida's east coast. The current study assessed the dynamics of microcystins and saxitoxin in six segments of the IRL: Banana River Lagoon (BRL), Mosquito Lagoon (ML), Northern IRL (NIRL), Central IRL (CIRL), Southern IRL (SIRL), and the St. Lucie Estuary (SLE). Surface water samples (n = 40) collected during the 2018 wet and 2019 dry season were analyzed to determine associations between toxins and temperature, salinity, pH, oxygen saturation, concentrations of dissolved nutrients and chlorophyll-a, presence of biosynthetic genes for toxins, relative abundance of planktonic species, and composition of the microbial community. The potential toxicity of samples was assessed using multiple mammalian cell lines. Enzyme-Linked Immunosorbent Assays were used to determine concentrations of microcystins and saxitoxin. Overall, the microcystins concentration ranged between 0.01-85.70 µg/L, and saxitoxin concentrations ranged between 0.01-2.43 µg/L across the IRL. Microcystins concentrations were 65% below the limit of quantification (0.05 µg/L), and saxitoxin concentrations were 85% below the limit of detection (0.02 µg/L). Microcystins concentrations were higher in the SLE, while saxitoxin was elevated in the NIRL and BRL. Cytotoxicity related to the presence of microcystins was seen in the SLE during the wet season. No significant patterns between cytotoxicity and saxitoxin were identified. Dissolved nutrients were identified as the most highly related parameters, explaining 53% of microcystin and 47% of saxitoxin variability. Multivariate models suggested cyanobacteria, flagellates, ciliates, and diatoms as the subset of microorganisms whose abundances were maximally correlated with saxitoxin and microcystins concentrations. Lastly, biosynthetic genes for microcystins were detected in the SLE and for saxitoxin in the BRL and NIRL. These results highlight the synergistic roles environmental and biological parameters play in influencing the dynamics of toxin production by harmful algae in the IRL.


Assuntos
Microcistinas , Rios , Animais , Florida , Saxitoxina
19.
Chemosphere ; 278: 130224, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-33813339

RESUMO

The scallop, Patinopecten yessoensis, was screened for new saxitoxin analogues to study the metabolism of paralytic shellfish toxins (PSTs), and this resulted in the discovery of two new analogues: M5-hemiaminal (HA) and M6-HA. M5-HA was isolated and its structure was determined by using NMR spectroscopy. It contains hydrogen at C-4 with opposite stereochemistry to that in saxitoxin, and a hemiaminal was formed between 9-NH2 and the hydrated ketone at C-12 in α-orientation. This is the first reported structural feature in a natural saxitoxin analogue, whereas the same ring system has previously been reported in a synthetic saxitoxin analogue, FD-saxitoxin. Acid hydrolysis of the carbamoyl N-sulfate in M5-HA produced M6-HA which was also identified in P. yessoensis by using LC-MSMS. M5-HA was not synthetically produced from M1 (11-hydroxy gonyautoxin-5) and M3 (11,11-dihydroxy gonyautoxin-5) through incubation in aqueous buffers. Furthermore, PSTs in the hepatopancreas of P. yessoensis, cultured in a bay located in northeastern Japan, were chronologically analyzed in 2018. The highest concentrations of M1/M3/M5-HA were observed two weeks after C-toxins had reached their highest concentrations, which provides evidence that M1/M3/M5-HA are metabolites of C-toxins. The voltage-gated sodium channel blockage activity of M6-HA was not detected at the concentration of 140 nM by using the Neuro-2A veratridine/ouabain assay.


Assuntos
Pectinidae , Saxitoxina , Animais , Japão , Saxitoxina/toxicidade , Alimentos Marinhos , Frutos do Mar/análise
20.
Sensors (Basel) ; 21(7)2021 Apr 03.
Artigo em Inglês | MEDLINE | ID: mdl-33916687

RESUMO

In the past twenty years marine biotoxin analysis in routine regulatory monitoring has advanced significantly in Europe (EU) and other regions from the use of the mouse bioassay (MBA) towards the high-end analytical techniques such as high-performance liquid chromatography (HPLC) with tandem mass spectrometry (MS). Previously, acceptance of these advanced methods, in progressing away from the MBA, was hindered by a lack of commercial certified analytical standards for method development and validation. This has now been addressed whereby the availability of a wide range of analytical standards from several companies in the EU, North America and Asia has enhanced the development and validation of methods to the required regulatory standards. However, the cost of the high-end analytical equipment, lengthy procedures and the need for qualified personnel to perform analysis can still be a challenge for routine monitoring laboratories. In developing regions, aquaculture production is increasing and alternative inexpensive Sensitive, Measurable, Accurate and Real-Time (SMART) rapid point-of-site testing (POST) methods suitable for novice end users that can be validated and internationally accepted remain an objective for both regulators and the industry. The range of commercial testing kits on the market for marine toxin analysis remains limited and even more so those meeting the requirements for use in regulatory control. Individual assays include enzyme-linked immunosorbent assays (ELISA) and lateral flow membrane-based immunoassays (LFIA) for EU-regulated toxins, such as okadaic acid (OA) and dinophysistoxins (DTXs), saxitoxin (STX) and its analogues and domoic acid (DA) in the form of three separate tests offering varying costs and benefits for the industry. It can be observed from the literature that not only are developments and improvements ongoing for these assays, but there are also novel assays being developed using upcoming state-of-the-art biosensor technology. This review focuses on both currently available methods and recent advances in innovative methods for marine biotoxin testing and the end-user practicalities that need to be observed. Furthermore, it highlights trends that are influencing assay developments such as multiplexing capabilities and rapid POST, indicating potential detection methods that will shape the future market.


Assuntos
Toxinas Marinhas , Saxitoxina , Animais , Ásia , Europa (Continente) , Camundongos , Ácido Okadáico
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