Saxitoxin potentiates human neuronal cell death induced by Zika virus while sparing neural progenitors and astrocytes.

The Zika virus (ZIKV) epidemic declared in Brazil between 2015 and 2016 was associated with an increased prevalence of severe congenital malformations, including microcephaly. The distribution of microcephaly cases was not uniform across the country, with a disproportionately higher incidence in the Northeast region (NE). Our previous work demonstrated that saxitoxin (STX), a toxin present in the drinking water reservoirs of the NE, exacerbated the damaging effects of ZIKV on the developing brain. We hypothesized that the impact of STX might vary among different neural cell types. While ZIKV infection caused severe damages on astrocytes and neural stem cells (NSCs), the addition of STX did not exacerbate these effects. We observed that neurons subjected to STX exposure were more prone to apoptosis and displayed higher ZIKV infection rate. These findings suggest that STX exacerbates the harmful effects of ZIKV on neurons, thereby providing a plausible explanation for the heightened severity of ZIKV-induced congenital malformations observed in Brazil's NE. This study highlights the importance of understanding the interactive effects of environmental toxins and infectious pathogens on neural development, with potential implications for public health policies.

Sublethal neurotoxicity of saxitoxin in early zebrafish development: Impact on sensorimotor function and neurotransmission systems.

Saxitoxin (STX) represents a marine toxin of significant concern due to its deleterious implications for aquatic ecosystems and public food safety. As a potent paralytic agent, the role of STX in obstructing voltage-gated sodium channels (VGSCs) is well-characterized. Yet, the mechanistic details underlying its low-dose toxicity remain largely enigmatic. In the current study, zebrafish embryos and larvae were subjected to subchronic exposure of graded STX concentrations (0, 1, 10, and 100 µg/L) until the 7th day post-fertilization. A tactile stimulus-based assay was employed to evaluate potential behavioral perturbations resulting from STX exposure. Both behavioral and transcription level analyses unveiled a compromised tactile response, which was found to be associated with a notable upregulation in the mRNA of two distinct VGSC isoforms, specifically the scn8aa/ab and scn1Laa/ab transcripts, even at the minimal STX dose. Notably, exposure to this lowest STX concentration also resulted in alterations in the transcriptional patterns of pivotal genes for cholinergic and GABAergic pathways, including ache and gabra1. Furthermore, STX induced a marked decrease in the levels of the neurotransmitter GABA. Our findings underscore that prolonged low-dose STX exposure during early development can significantly compromise the tactile response behavior in zebrafish. This study reveals that chronic low-dose STX exposure of developing zebrafish alters neurotransmission pathways that converge on altered tactile behavior.

Growth and paralytic shellfish poisoning toxin production by a Mexican dinoflagellate strain of Alexandrium tamiyavanichii Balech (1994) under different nutrient conditions.

Alexandrium tamiyavanichii is a marine dinoflagellate known to produce Paralytic Shellfish Poisoning (PSP) toxin. Thus, a strain was isolated from La Paz Bay, Baja California Sur, Mexico and used to explore whether stress conditions, such as phosphorus limitation (PL) and nitrogen enrichment (NE) modulate population growth and PSP toxin production in the GSe medium. Growth kinetics showed that the PL treatment produced a 3.4-fold increase in cell density versus control at day 30 of the culture cycle. The highest PSP concentration was found in the control culture (309 fmol cell-1) on day 21. Saxitoxin (STX) was the main analog in all the treatments (> 40 % mol). In conclusion, PL and NE treatments promoted growth kinetics in the species studied but did not affect the PSP toxin production. For the first time, the present research describes A. tamiyavanichii high toxicity strain isolated from Mexican coasts relative to the South-Atlantic strains.

Toxicogenomic Effects of Dissolved Saxitoxin on the Early Life Stages of the Longfin Yellowtail (Seriola rivoliana).

Harmful algal blooms (HABs) can produce a variety of noxious effects and, in some cases, the massive mortality of wild and farmed marine organisms. Some HAB species produce toxins that are released into seawater or transferred via food webs (particulate toxin fraction). The objective of the present study was to identify the toxicological effects of subacute exposure to saxitoxin (STX) during embryonic and early larval stages in Seriola rivoliana. Eggs were exposed to dissolved 19 STX (100 µg L-1). The toxic effects of STX were evaluated via the hatching percentage, the activity of three enzymes (protein and alkaline phosphatases and peroxidase), and the expression of four genes (HSF2, Nav1.4b, PPRC1, and DUSP8). A low hatching percentage (less than 5%) was observed in 44 hpf (hours post fertilization) embryos exposed to STX compared to 71% in the unexposed control. At this STX concentration, no oxidative stress in the embryos was evident. However, STX induced the expression of the NaV1.4 channel α-subunit (NaV1.4b), which is the primary target of this toxin. Our results revealed the overexpression of all four candidate genes in STX-intoxicated lecithotrophic larvae, reflecting the activation of diverse cellular processes involved in stress responses (HSF2), lipid metabolism (PPRC1), and MAP kinase signaling pathways associated with cell proliferation and differentiation (DUSP8). The effects of STX were more pronounced in young larvae than in embryos, indicating a stage-specific sensitivity to the toxin.

A New Bioassay for the Detection of Paralytic and Amnesic Biotoxins Based on Motor Behavior Impairments of Zebrafish Larvae.

The global concern about the increase of harmful algal bloom events and the possible impacts on food safety and aquatic ecosystems presents the necessity for the development of more accessible techniques for biotoxin detection for screening purposes. Considering the numerous advantages that zebrafish present as a biological model and particularly as a toxicants sentinel, we designed a sensitive and accessible test to determine the activity of paralytic and amnesic biotoxins using zebrafish larvae immersion. The ZebraBioTox bioassay is based on the automated recording of larval locomotor activity using an IR microbeam locomotion detector, and manual assessment of four complementary responses under a simple stereoscope: survival, periocular edema, body balance, and touch response. This 24 h acute static bioassay was set up in 96-well microplates using 5 dpf zebrafish larvae. For paralytic toxins, a significant decrease in locomotor activity and touch response of the larvae was detected, allowing a detection threshold of 0.1-0.2 µg/mL STXeq. In the case of the amnesic toxin the effect was reversed, detecting hyperactivity with a detection threshold of 10 µg/mL domoic acid. We propose that this assay might be used as a complementary tool for environmental safety monitoring.

Paralytic Shellfish Toxins of Pyrodinium bahamense (Dinophyceae) in the Southeastern Gulf of Mexico.

In September and November 2016, eight marine sampling sites along the coast of the southeastern Gulf of Mexico were monitored for the presence of lipophilic and hydrophilic toxins. Water temperature, salinity, hydrogen potential, dissolved oxygen saturation, inorganic nutrients and phytoplankton abundance were also determined. Two samples filtered through glass fiber filters were used for the extraction and analysis of paralytic shellfish toxins (PSTs) by lateral flow immunochromatography (IFL), HPLC with post-column oxidation and fluorescent detection (FLD) and UHPLC coupled to tandem mass spectrometry (UHPLC-MS/MS). Elevated nutrient contents were associated with the sites of rainwater discharge or those near anthropogenic activities. A predominance of the dinoflagellate Pyrodinium bahamense was found with abundances of up to 104 cells L-1. Identification of the dinoflagellate was corroborated by light and scanning electron microscopy. Samples for toxins were positive by IFL, and the analogs NeoSTX and STX were identified and quantified by HPLC-FLD and UHPLC-MS/MS, with a total PST concentration of 6.5 pg cell-1. This study is the first report that confirms the presence of PSTs in P. bahamense in Mexican waters of the Gulf of Mexico.

Evaluation of Paralytic Shellfish Toxins in Marine Oyster Farming and Microalgae in the Atlantic Amazon Evidences Safety but Highlights Potential Risks of Shellfish Poisoning.

Marine phycotoxins are organic compounds synthesized by some species of microalgae, which accumulate in the tissues of filter-feeder organisms such as bivalve mollusks. These toxins can cause acute intoxication episodes in humans, a severe threat to aquaculture and fisheries. In the State of Pará, Brazil, oyster farming has community, artisanal and sustainable bases, using mangroves as cultivation environment and seed banks. In small-scale production, there are often no established methods of safeguarding the health of consumers elevating the potential risks of shellfish poisoning outbreaks. Our study evaluated the presence of phycotoxins in oysters cultivated in five municipalities in the region of the Atlantic Amazon (Pará, Brazil) assessing the quality of the final product. We further evaluated the microalgae, water quality, and the spatio-temporal variation of physicochemical factors in the same area. Diatoms dominated the microalgae composition, followed by dinoflagellates, some of which are reported to be potentially toxic and producers of paralytic shellfish toxins. For the first time, we describe the occurrence of the potentially toxic dinoflagellate Ostreopsis sp. in the Amazon region. Furthermore, for the first time, toxins were detected in oyster farming in the northeast of the State of Pará, namely GTX2,3, STX, and dc-STX nevertheless, with nontoxic values. The identified toxins represent a potential threat to shellfish consumers.

Saxitoxin Group Toxins Accumulation Induces Antioxidant Responses in Tissues of Mytilus chilensis, Ameghinomya antiqua, and Concholepas concholepas during a Bloom of Alexandrium pacificum.

Saxitoxin (STX) group toxins consist of a set of analogues which are produced by harmful algal blooms (HABs). During a HAB, filter-feeding marine organisms accumulate the dinoflagellates and concentrate the toxins in the tissues. In this study, we analyze the changes in antioxidant enzymes and oxidative damage in the bivalves Mytilus chilensis and Ameghinomya antiqua, and the gastropod Concholepas concholepas during a bloom of Alexandrium pacificum. The results show that during the exponential phase of the bloom bivalves show an increase in toxicity and activity of antioxidant enzymes (superoxide dismutase, catalase, glutathione peroxidase, and glutathinoe reductase, p < 0.05), while in the gastropods, increased activity of antioxidant enzymes was associated with the bioaccumulation of toxins through the diet. At the end of the bloom, decreased activity of antioxidant enzymes in the visceral and non-visceral tissues was detected in the bivalves, with an increase in oxidative damage (p < 0.05), in which the latter is correlated with the detection of the most toxic analogues of the STX-group (r = 0.988). In conclusion, in areas with high incidence of blooms, shellfish show a high activity of antioxidants, however, during the stages involving the distribution and bioconversion of toxins, there is decreased activity of antioxidant enzymes resulting in oxidative damage.

Permanent occurrence of Raphidiopsis raciborskii and cyanotoxins in a subtropical reservoir polluted by domestic effluents (Itupararanga reservoir, São Paulo, Brazil).

Toxic cyanobacteria blooms are a frequent problem in subtropical reservoirs and freshwater systems. The purpose of this study was to investigate the occurrence of potentially toxic cyanobacteria and the environmental conditions associated with the presence of cyanotoxins in a Brazilian subtropical reservoir. Five collections were carried out at seven sampling locations in the reservoir, during the rainy and dry seasons, between the years 2016 and 2017. There was permanent occurrence of Raphidiopsis raciborskii (Woloszynska) Aguilera, Berrendero Gómez, Kastovsky, Echenique & Salerno (Phycologia 57(2):130-146, 2018), ranging between dominant and abundant, with an average biomass of 38.8 ± 29.9 mg L-1. Also abundant were Dolichospermum solitarium, D. planctonicum, Planktothrix isothrix, and Aphanizomenon gracile. Saxitoxin (STX) was detected in all the collected samples (0.11 ± 0.05 µg L-1). Microcystin (MC) was also detected, but at lower concentrations (0.01 ± 0.0 µg L-1). Low availability of NO3- and phosphorus limitation had significant effects on the R. raciborskii biomass and the levels of STX and MC. It was observed that R. raciborskii was sensitive to thermal stratification, at the same time that STX levels were higher. This suggested that STX was produced under conditions that restricted the growth of R. raciborskii. These are important findings, because they add information about the permanent occurrence of STX and R. raciborskii in an aquatic ecosystem limited by phosphorus, vulnerable to climatic variations, and polluted by domestic effluents.

Electrochemical impedance biosensor for detection of saxitoxin in aqueous solution.

Saxitoxin is a cyanotoxin which is very harmful to human health; the concentration limit in drinking water is only 3 µg/L. Therefore, a simple, fast, sensitive, low-cost, and specific method for its detection, quantification, and monitoring in water bodies is needed to avoid adverse effects on animal and human health. In this work, we developed an electrochemical impedimetric biosensor using a specific aptamer as recognition element for saxitoxin detection. This method allies the superior sensing characteristics of aptamers with the nondestructive, label-free, and easy working principles of the electrochemical impedance technique. The device presented sensitivity for detecting saxitoxin concentrations above 0.3 µg/L, with high selectivity in negative control experiments, demonstrating a promising alternative for water toxin detection.

Effects of paralytic shellfish toxins on the middle intestine of Oncorhynchus mykiss: Glutathione metabolism, oxidative status, lysosomal function and ATP-binding cassette class C (ABCC) proteins activity.

We studied the absorption, cytotoxicity and oxidative stress markers of Paralytic Shellfish Toxins (PST) from three extracts from Alexandrium catenella and A. ostenfeldii, in middle Oncorhynchus mykiss intestine in vitro and ex vivo preparations. We measured glutathione (GSH) content, glutathione-S transferase (GST), glutathione reductase (GR) and catalase (CAT) enzymatic activity, and lipid peroxidation in isolated epithelium exposed to 0.13 and 1.3 µM PST. ROS production and lysosomal membrane stability (as neutral red retention time 50%, NRRT50) were analyzed in isolated enterocytes exposed to PST alone or plus 3 µM of the ABCC transport inhibitor MK571. In addition, the concentration-dependent effects of PST on NRRT50 were assayed in a concentration range from 0 to 1.3 µM PST. We studied the effects of three different PST extracts on the transport rate of the ABCC substrate DNP-SG by isolated epithelium. The extract with highest inhibition capacity was selected for studying polarized DNP-SG transport in everted and non-everted intestinal segments. We registered lower GSH content and GST activity, and higher GR activity, with no significant changes in CAT activity, lipid peroxidation or ROS level. PST exposure decreased NRRT50 in a concentration-depend manner (IC50 = 0.0045 µM), but PST effects were not augmented by addition of MK571. All the three PST extracts inhibited ABCC transport activity, but this inhibition was effective only when the toxins were applied to the apical side of the intestine and DNP-SG transport was measured at the basolateral side. Our results indicate that PST are absorbed by the enterocytes from the intestine lumen. Inside the enterocytes, these toxins decrease GSH content and inhibit the basolateral ABCC transporters affecting the normal functions of the cell. Furthermore, PST produce a strong cytotoxic effect to the enterocytes by damaging the lysosomal membrane, even at low, non-neurotoxic concentrations.

Toxigenic phytoplankton groups and neurotoxin levels related to two contrasting environmental conditions at the coastal area of Rio de Janeiro (west of South Atlantic).

An assessment of the major pigments and neurotoxins and a description of the phytoplankton community were carried out within the coastal region of Rio de Janeiro State (Brazil), during winter and the following spring of 2018. Overall, six stations were investigated for oceanographic conditions (with CTD casts). Filtered water samples were used to estimate the chlorophyll a (CHL-a), carotenoids (CAR), and phycobiliproteins (PHY) using UV-Vis spectrophotometry, as well as the quantification of saxitoxins (STX) and domoic acid (DA), through High Performance Liquid Chromatography (HPLC). Planktonic organisms were counted using sedimentation chambers of different volumes and an inverted microscope. A cluster analysis, SIMPER, and ANOSIM were applied to the phytoplankton data along with diversity indexes, and non-parametric statistics to phycotoxins and pigments. There was a significant difference between the winter and spring phytoplankton community, associated with the mixed layer depth (r2 = -0.626, p < 0.05) and temperature (r2 = 0.641, p < 0.05). Phytoplankton biomass and C:CHL-a indicated a higher production during the winter than in spring, with the potentially toxic genus Pseudo-nitzschia responsible for 12.79% of autotrophic abundance (SIMPER output). Pigments showed a slight increase in CAR during spring, while PHY remained at trace concentrations. Both the DA and STX were quantified in winter and spring, but with significant differences only for STX between the sampling periods. Among the 71 taxa, 11 were identified as potentially toxic with an emphasis on STX-producing dinoflagellates and cyanobacteria, such as Alexandrium sp., Gymnodinium spp. along with Trichodesmium spp. Season-related environmental variability may be the major driving force modulating the mixed assemblage of species that support different levels of phycotoxins.

Rapid screening fluorescence method applied to detection and quantitation of paralytic shellfish toxins in invertebrate marine vectors.

A rapid screening method is described for the determination of paralytic shellfish toxins (PST), in fresh marine vectors (bivalves and gastropods), at levels ranging from 0.05 to 5.0 mg STX-eq kg-1. PST are extracted from marine vector homogenates with acetic acid according to the Pre-COX-LC-FLD method. At the same time, the obtained extract is oxidised simultaneously in hydrogen peroxide and periodate oxidate to determine PST, non-N-hydroxylated and N-hydroxylated toxins, respectively. Then, they are analysed using a microplate fluorometer (Ex: 335 nm/Em: 405 nm). All the samples were compared with the liquid chromatography post-column oxidation method. Recoveries of PST added to fresh and processed marine vectors averaged 93.9% with a coefficient of variation of 6.1%. Both methods showed a good linear regression (r2 = 0.97). The method shows good intra- and inter-day precisions with a relative coefficient of variation of ≈ 3.8% and 5.7%, respectively. The limit of quantification of the rapid screening fluorescence method was ≈ 0.082 mg STX-eq kg-1, with ≤5% false positives. The established rapid screening fluorescence methods offer highly effective and verifiable pre-analyses of PST contamination in marine vectors and can be used for routine screening of the PST in seafood before formal identification by confirmatory methods (Pre-COX LC-FLD method, Lawrence method).

Accumulation and biotransformation dynamics of the neurotoxic complex, saxitoxin, in different life stages of Ostrea chilensis.

The neurotoxic complex saxitoxin, is a group of marine toxins that historically has significantly impacted human health and the ability to utilize marine resources. A steady increase in the distribution and intensity of Alexandrium catenella blooms in Chile, and around the world, has caused major ecological and socioeconomic impacts, putting this type of dinoflagellate, and its toxicity, in the spotlight. Ostrea chilensis is a commercially and ecologically important resource harvested from wild populations and farmed in centers of southern Chile, where it is exposed to large harmful algal blooms of the type that can cause paralysis in humans. This study contributes to our understanding about the transfer of toxins from A. catenella cells to juvenile and adult Ostrea chilensis by tracking transformations of the neurotoxic complex until it reaches its most stable molecular form in the intracellular environment of O. chilensis tissues. These biotransformations are different in O. chilensis juveniles and adults, indicating a differentiated response for these two life stages of this bivalve species. These studies can be used for similar analyses in other ecologically and commercially important species of filter feeding organisms, providing greater understanding of the specific interactions of bivalves in scenarios of toxic dinoflagellate proliferations (e.g. A. catenella blooms).

Effects of increased zooplankton biomass on phytoplankton and cyanotoxins: A tropical mesocosm study.

Zooplankton are important biocontrol agents for algal blooms in temperate lakes, while their potential in tropical and subtropical environments is not well understood. The aim of the present study was to evaluate the influence of increased zooplankton biomass on phytoplankton community and cyanotoxins (microcystins and saxitoxin) content of a tropical reservoir (Ipojuca reservoir, Brazil) using in situ mesocosms. Mesocosms consisted of 50L transparent polyethylene bags suspended in the reservoir for twelve days. Phytoplankton populations were exposed to treatments having 1 (control), 2, 3 and 4 times the biomass of zooplankton found in the reservoir at the beginning of the experiment. Filamentous cyanobacteria such as Planktothrix agardhii and Cylindrospermopsis raciborskii were not negatively influenced by increasing zooplankton biomass. In contrast, the treatments with 3 and 4 times zooplankton biomass negatively affected the cyanobacteria Aphanocapsa sp., Chroococcus sp., Dolichospermum sp., Merismopedia tenuissima, Microcystis aeruginosa and Pseudanabaena sp.; the diatom Cyclotella meneghiniana; and the cryptophyte Cryptomonas sp. Total microcystin concentration both increased and decreased at different times depending on zooplankton treatment, while saxitoxin level was not significantly different between the treatments and control. The results of the present study suggest that zooplankton biomass can be manipulated to control the excessive proliferation of non-filamentous bloom forming cyanobacteria (e.g. M. aeruginosa) and their associated cyanotoxins.

Guanidinium Toxins and Their Interactions with Voltage-Gated Sodium Ion Channels.

Guanidinium toxins, such as saxitoxin (STX), tetrodotoxin (TTX) and their analogs, are naturally occurring alkaloids with divergent evolutionary origins and biogeographical distribution, but which share the common chemical feature of guanidinium moieties. These guanidinium groups confer high biological activity with high affinity and ion flux blockage capacity for voltage-gated sodium channels (NaV). Members of the STX group, known collectively as paralytic shellfish toxins (PSTs), are produced among three genera of marine dinoflagellates and about a dozen genera of primarily freshwater or brackish water cyanobacteria. In contrast, toxins of the TTX group occur mainly in macrozoa, particularly among puffer fish, several species of marine invertebrates and a few terrestrial amphibians. In the case of TTX and analogs, most evidence suggests that symbiotic bacteria are the origin of the toxins, although endogenous biosynthesis independent from bacteria has not been excluded. The evolutionary origin of the biosynthetic genes for STX and analogs in dinoflagellates and cyanobacteria remains elusive. These highly potent molecules have been the subject of intensive research since the latter half of the past century; first to study the mode of action of their toxigenicity, and later as tools to characterize the role and structure of NaV channels, and finally as therapeutics. Their pharmacological activities have provided encouragement for their use as therapeutants for ion channel-related pathologies, such as pain control. The functional role in aquatic and terrestrial ecosystems for both groups of toxins is unproven, although plausible mechanisms of ion channel regulation and chemical defense are often invoked. Molecular approaches and the development of improved detection methods will yield deeper understanding of their physiological and ecological roles. This knowledge will facilitate their further biotechnological exploitation and point the way towards development of pharmaceuticals and therapeutic applications.

Hierarquização da eficiência de remoção de cianotoxinas por meio de adsorção em carvão ativado granular / Evaluation of hierarchy of cyanotoxin removal by adsorption onto granular activated carbon

RESUMO O trabalho objetivou avaliar a remoção concomitante de duas cianotoxinas - microcistina-LR e saxitoxina-STX - por meio da adsorção em carvão ativado granular. Em primeira etapa, três carvões de granulometrias distintas, dois de coco de dendê e um mineral, foram avaliados na adsorção de microcistina-LR em água destilada e tratada com concentrações de 3,9 a 9,4 ug.L-1. O carvão de coco de dendê de menor granulometria apresentou desempenho mais uniforme, tanto no que tange à remoção (%) de microcistina-LR, quanto ao volume de efluente produzido consoante com as recomendações do padrão de potabilidade brasileiro. Em segunda etapa, apenas para o carvão selecionado na etapa precedente, concentrações de microcistina-LR e saxitoxina-STX aproximadamente 50% superiores dos limites do mesmo padrão de potabilidade foram dispersas em água destilada. Tais ensaios de adsorção evidenciaram que apenas a adsorção de saxitonina não foi influenciada pela simultaneidade das duas cianotoxinas.

ABSTRACT This study focused on the simultaneous removal of two cyanotoxins - microcystin-LR and saxitoxin-STX - by means of adsorption onto granular activated carbon. In first step, three carbons with different granulometry (dende coconut and mineral) were evaluated on adsorption of microcystin-LR in distilled and treated waters in concentrations at about 3.9 to 9.4 ug.L-1. The dende coconut carbon with lower granulometry showed higher performance according to the limits established by Brazilian Drinking Water Standards. In second step, with the selected carbon in previous step, concentrations of the two cyanotoxins 50% higher than those limits were dispersed in distilled water. The tests pointed out that only saxitoxin-STX removal was not affect by the presence both cyanotoxins.

Prevalence, Variability and Bioconcentration of Saxitoxin-Group in Different Marine Species Present in the Food Chain.

The saxitoxin-group (STX-group) corresponds to toxic metabolites produced by cyanobacteria and dinoflagellates of the genera Alexandrium, Gymnodinium, and Pyrodinium. Over the last decade, it has been possible to extrapolate the areas contaminated with the STX-group worldwide, including Chile, a phenomenon that has affected ≈35% of the Southern Pacific coast territory, generating a high economic impact. The objective of this research was to study the toxicity of the STX-group in all aquatic organisms (bivalves, algae, echinoderms, crustaceans, tunicates, cephalopods, gastropods, and fish) present in areas with a variable presence of harmful algal blooms (HABs). Then, the toxic profiles of each species and dose of STX equivalents ingested by a 60 kg person from 400 g of shellfish were determined to establish the health risk assessment. The toxins with the highest prevalence detected were gonyautoxin-4/1 (GTX4/GTX1), gonyautoxin-3/2 (GTX3/GTX2), neosaxitoxin (neoSTX), decarbamoylsaxitoxin (dcSTX), and saxitoxin (STX), with average concentrations of 400, 2800, 280, 200, and 2000 µg kg-1 respectively, a species-specific variability, dependent on the evaluated tissue, which demonstrates the biotransformation of the analogues in the trophic transfer with a predominance of α-epimers in all toxic profiles. The identification in multiple vectors, as well as in unregulated species, suggests that a risk assessment and risk management update are required; also, chemical and specific analyses for the detection of all analogues associated with the STX-group need to be established.

Interaction of single-walled carbon nanotubes and saxitoxin: Ab initio simulations and biological responses in hippocampal cell line HT-22.

Saxitoxins (STXs) are potent neurotoxins that also induce cytotoxicity through the generation of reactive oxygen species. Carbon nanotubes (CNTs) are nanomaterials that can promote a Trojan horse effect, facilitating the entry of toxic molecules to cells when adsorbed to nanomaterials. The interaction of pristine single-walled (SW)CNTs and carboxylated (SWCNT-COOH) nanotubes with STX was evaluated by ab initio simulation and bioassays using the cell line HT-22. Cells (5 × 104 cells/mL) were exposed to SWCNT and SWCNT-COOH (5 µg mL-1 ), STX (200 µg L-1 ), SWCNT+STX, and SWCNT-COOH+STX for 30 min or 24 h. Results of ab initio simulation showed that the interaction between SWCNT and SWCNT-COOH with STX occurs in a physisorption. The interaction of SWCNT+STX induced a decrease in cell viability. Cell proliferation was not affected in any treatment after 30 min or 24 h of exposure (p > 0.05). Treatment with SWCNT-COOH induced high reactive oxygen species levels, an effect attenuated in SWCNT-COOH+STX treatment. In terms of cellular oxygen consumption, both CNTs when coexposed with STX antagonize the toxin effect. Based on these results, it can be concluded that the results obtained in vitro corroborate the semiempirical evidence found using density functional theory ab initio simulation. Environ Toxicol Chem 2017;36:1728-1737. © 2016 SETAC.

Is qPCR a Reliable Indicator of Cyanotoxin Risk in Freshwater?

The wide distribution of cyanobacteria in aquatic environments leads to the risk of water contamination by cyanotoxins, which generate environmental and public health issues. Measurements of cell densities or pigment contents allow both the early detection of cellular growth and bloom monitoring, but these methods are not sufficiently accurate to predict actual cyanobacterial risk. To quantify cyanotoxins, analytical methods are considered the gold standards, but they are laborious, expensive, time-consuming and available in a limited number of laboratories. In cyanobacterial species with toxic potential, cyanotoxin production is restricted to some strains, and blooms can contain varying proportions of both toxic and non-toxic cells, which are morphologically indistinguishable. The sequencing of cyanobacterial genomes led to the description of gene clusters responsible for cyanotoxin production, which paved the way for the use of these genes as targets for PCR and then quantitative PCR (qPCR). Thus, the quantification of cyanotoxin genes appeared as a new method for estimating the potential toxicity of blooms. This raises a question concerning whether qPCR-based methods would be a reliable indicator of toxin concentration in the environment. Here, we review studies that report the parallel detection of microcystin genes and microcystin concentrations in natural populations and also a smaller number of studies dedicated to cylindrospermopsin and saxitoxin. We discuss the possible issues associated with the contradictory findings reported to date, present methodological limitations and consider the use of qPCR as an indicator of cyanotoxin risk.