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Contributing to the assessment of potential physiological changes in microalgae subjected to different concentrations and types of cyanotoxins, this study investigated the inhibitory effects of cyanotoxins on the growth, density, biomass, and ecotoxicity of Chlorella vulgaris. Chlorella vulgaris was exposed to crude extracts of cyanobacteria producing microcystin-LR (MC-LR), saxitoxin (SXT), anatoxin-a (ATX-A), and cylindrospermopsin (CYN) with initial concentrations of 5.0, 2.05, 0.61, and 1.42 μg.L−1, respectively. The experiments were conducted under controlled conditions, and monitoring of growth and cell inhibition occurred at 24h, 48h, 72h, and 96h. Chlorophyll-a content and ecotoxicity assessment were conducted with samples collected after 96h of exposure to cyanotoxins. The growth assays of Chlorella vulgaris , with results expressed in terms of average growth rates (doublings/day), indicated the following order for cyanotoxins: SXT (2.03) > CYN (1.66) > MC-LR (1.56) > ATX-A (0.18). This assay revealed the prominent inhibitory potential of ATX-A on Chlorella vulgaris growth compared to the other toxins evaluated. Regarding the inhibition of the photosynthetic process, expressed in terms of the percentage inhibition of Chlorophyll-a, the following order for cyanotoxins was obtained: ATX-A (82%) > MC-LR (76%) > STX (46%) > CYN (16%). These results also indicated that among the cyanotoxins, ATX-A was the most detrimental to the photosynthetic process. However, contrary to the observations in the growth study, SXT proved to be more harmful than CYN in terms of Chlorophyll-a inhibition. Finally, the results of the toxicity assay revealed that only ATX-A and MC-LR exerted a chronic influence on Chlorella vulgaris under the investigated conditions. [Display omitted] • Phytotoxicity of cyanotoxins induces different physiological responses to Chlorella vulgaris. • MC-LR and ATX-A promote greater inhibitory effects on the growth, density, biomass and ecotoxicity of Chlorella vulgaris. • The lowest inhibitions of algal growth among the cyanotoxins were for SXT and CYN. • The percentage inhibition of Chlorophyll –a was: ATX-A (82%) > MC-LR (76%) > SAX (46%) > CYN (16%). [ABSTRACT FROM AUTHOR]

For food safety, the concentrations and profiles of paralytic shellfish toxins (PSTs) and tetrodotoxin were examined in economically important scallops and bloody clams collected from the coast of the Miyagi Prefecture, Japan. PSTs were the major toxins in both species. The tetrodotoxin concentration in scallops increased in summer, although the highest value (18.7 μg/kg) was lower than the European Food Safety Authority guideline threshold (44 μg/kg). This confirmed the safety for tetrodotoxin in this area. [Display omitted] • Paralytic shellfish toxins and tetrodotoxin in scallops and bloody clams from Miyagi Prefecture, Japan, were analyzed. • Paralytic shellfish toxins were the major toxic components in both bivalve species. • The composition of paralytic shellfish toxins showed characteristic differences between the two species. • In scallops, the concentration of tetrodotoxin reached its highest level in late August. • Highest concentration of tetrodotoxin in scallops was lower than the European Food Safety Authority guideline threshold. [ABSTRACT FROM AUTHOR]

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. [Display omitted] • Enzymatic production of decarbamoyl gonyautoxin 1&4. • Purification of equilibrated epimeric mixture of decarbamoyl gonyautoxin 1&4. • Acute toxicity of decarbamoyl gonyautoxin 1&4 in mice by i.p. injection and feeding. • Possible metabolism of decarbamoyl gonyautoxin 1&4 after feeding to mice. [ABSTRACT FROM AUTHOR]

In this study, we proposed label-free saxitoxin (STX) sensor using STX specific aptamer in combination with spectroscopic ellipsometry (SE) and attenuated internal reflection (AIR) spectroscopic ellipsometry method which is operated under surface plasmon resonance (SPR) conditions. Besides the other surface plasmon resonance-based applications, AIR-SE applications have unique advantages in terms of sensitivity and it was used herein for real-time detection of STX in real samples. Another method, SE, was also used and compared with AIR-SE. Analytical performances were satisfactory with low detection limits and a wide detection range. Limit of detection was 0.01 ng/mL for AIR-SE and 0.11 ng/mL for SE. Both proposed sensors were operable in 0.01 nM–1000 nM STX range. These methods were also used for the accurate, selective, and sensitive detection of STX from fish and shrimp samples. Image 1 • Selective and sensitive detection of saxitoxin from fishery products is reported here. • The first time, a total internal reflection spectrophotometric ellipsometry method is proposed for saxitoxin. • The limit of detection is 0.01 ng/mL which is under the permissible limits (80 ng/mL). [ABSTRACT FROM AUTHOR]

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. • Diatoms (Pseudo-nitzschia spp.) in high abundance at Rio de Janeiro coastal site. • Water column structure can influence phytoplankton community in winter and spring. • Saxitoxins and Domoic Acid levels changes with phytoplankton community composition. • Chlorophyll- a and carotenoids are affected by water column structure. • Carbon-chlorophyll a ratio: growth in winter and refractory carbon stock in spring. [ABSTRACT FROM AUTHOR]

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. • Solid Phase Adsorption Toxin Tracking used to track multiple algal toxins at seven locations in southern California. • Domoic acid, saxitoxin, okadaic acid co-occurred in 12% of samples collected. • First reported detection of dissolved okadaic acid in southern California waters. [ABSTRACT FROM AUTHOR]

To evaluate Neosaxitoxin (NeoSTX) as a local anesthetic drug, for pain control during and after piglet castration. Prospective, randomized and double-blind study. 24 commercial hybrids, males, 23-day-old piglets. The piglets were randomized into two groups: a Lidocaine group and a NeoSTX group. One minute before castration, they were injected intra-scrotally with a single dose of Lidocaine (20 mg, in 1 mL) and NeoSTX (0.1 μg, in 1 mL), respectively. NeoSTX does not generate vasoconstriction or scrotal contraction, unlike Lidocaine, where a decrease in temperature and scrotal size is observed within 5 min after the procedure. After 24 h, wound inflammation, as measured by scrotal size, was lower in the NeoSTX group. No significant difference could be shown between the vocalizations and facial expressions of pain of both groups during the castration procedure. A single dose of NeoSTX is safe and effective for pain management during and after piglet castration. NeoSTX treated piglets were less affected by castration than those in the Lidocaine group, thus reducing piglet stress and enhancing the quality of piglet convalescence. • Neosaxitoxin used as a local anesthetic during piglet castration. • A single dose of Neosaxitoxin is safe and effective for pain management. • Neosaxitoxin treated piglets were less affected by castration than those that had received Lidocaine. [ABSTRACT FROM AUTHOR]

Abstract To date Paralytic shellfish toxin (PST) variants in cyanobacteria have primarily been characterized using high performance liquid chromatography coupled with fluorescence detection. In this study we re-evaluated the PST profiles of five cyanobacterial cultures (Dolichospermum circinale AWQC131C, Aphanizomenon sp. NH-5, Raphidiopsis raciborskii T3, Scytonema cf. crispum CAWBG524 and CAWBG72) and one environmental sample (Microseria wollei) using hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry. A total of 35 different PST variants were detected. D. circinale contained the highest number of variants (23), followed by S. cf. crispum CAWBG72 (21). Many of the variants detected in the cultures/environmental sample had not been reported from these strains previously: D. circinale (14 variants), S. cf. crispum CAWBG72 (16), S. cf. crispum CAWBG524 (9), Aphanizomenon sp. (9), R. raciborskii (7), and M. wollei (7). Of particular interest was the detection of M-toxins (Aphanizomenon sp., R. raciborskii, D. circinale). These have previously only been identified from shellfish where they were thought to be metabolites. Well-characterized PST variant profiles are essential for research investigating the genetic basis of PST production, and given that the toxicity of each variants differs, it will assist in refining risk assessments. Highlights • Thirty-five paralytic shellfish toxin variants were detected in five cyanobacterial strains and one environmental sample. • Twenty of these variants had not been detected in these strains previously. • Dolichospermum circinale (AWQC131C) produced 23 variants. • M-toxins were detected of the first time in cyanobacteria. [ABSTRACT FROM AUTHOR]

Abstract The pufferfish saxitoxin- and tetrodotoxin-binding protein 2 (PSTBP2), which is involved in toxin accumulation, was knocked out in Takifugu rubripes embryos by using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 genome-editing technology. Treating the embryos with one of two single-guide RNA (sgRNA) resulted in mutation rates of 57.1% and 62.5%, respectively, as estimated using a heteroduplex mobility assay at 3 days postfertilization. Both sgRNAs might induced frameshift mutations that knocked out the T. rubripes PSTBP2. Highlights • The CRISPR/Cas9 system was used to knock out the PSTBP2 gene in Takifugu rubripes. • Off-target effects occurred at very low levels, or were not detected. • This knockout is useful for studying the mechanism of toxin accumulation in fishes. [ABSTRACT FROM AUTHOR]

Saxitoxin (STX) is a potent shellfish toxin found in freshwater and marine ecosystems which threatens human health by contaminating drinking water and shellfish. The formation of neutrophil extracellular traps (NETs) is a defense mechanism employed by polymorphonuclear leukocytes (PMNs) to destroy invading pathogens, and also plays a critical role in the pathogenesis of various diseases. In this study, we aimed to investigate the role of STX on human NET formation. Typical NETs-associated characteristics were detected from STX-stimulated PMNs using immunofluorescence microscopy. Moreover, NET quantification based on PicoGreen® fluorescent dye revealed that STX triggered NET formation in a concentration-dependent manner, and NET formation peaked at 120 min (with a total time of 180 min) after induction by STX. Intracellular reactive oxygen species (iROS) detection showed that iROS were significantly elevated in STX-challenged PMNs. These findings present insight into the effects of STX on human NET formation and serve as a basis for further investigations of STX immunotoxicity. [Display omitted] • Saxitoxin induced cell-free and anchored neutrophil extracellular traps release. • Saxitoxin triggered neutrophil extracellular traps in a concentration-dependent way. • Saxitoxin-induced neutrophil extracellular trap formation peaked at 120 min within a total assay time of 180 min. • Saxitoxin increased intracellular reactive oxygen species production in polymorphonuclear leukocytes. [ABSTRACT FROM AUTHOR]

A receptor binding assay (RBA) for the determination of paralytic shellfish poisoning toxicity is formally validated through collaborative study and approved for regulatory monitoring use in the US for mussels and clams. However, to date, the method has not been tested on bivalve molluscs originating from European waters and no validation studies have been conducted for oysters, a shellfish species of great importance globally. This study firstly reports the work conducted to assess the performance of the assay in comparison with a regulatory chemical detection method for a range of shellfish species originating from Great Britain. Data obtained showed a complete absence of false negative RBA results, with a tendency to over-estimate PSP toxicity for some shellfish species in comparison with liquid chromatography with fluorescence detection. Secondly, the performance of the RBA was assessed for oysters, with the analysis of a dilution series of oyster matrix certified reference materials. Method trueness, sensitivity and precision were found to compare well with results reported previously for other species. In addition, the RBA analysis of untreated and demetallated oyster extracts, provided good evidence that the RBA is not suppressed in the presence of high concentrations of zinc as reported previously for the mouse bioassay. Consequently, there is strong evidence from this study, that the RBA would be suitable for determination of PSP toxicity in bivalve molluscs from GB, with acceptable method performance in oysters. Further validation studies would be required for other shellfish species of interest before the method can be considered suitable for implementation in Europe. [ABSTRACT FROM AUTHOR]

Aphanizomenon gracile is one of the most widespread Paralytic Shellfish Toxin (PST) producing cyanobacteria in freshwater bodies in the Northern Hemisphere. It has been shown to produce various PST congeners, including saxitoxin (STX), neosaxitoxin (NEO), decarbamoylsaxitoxin (dcSTX) and gonyautoxin 5 (GTX5) in Europe, North America and Asia. Three cyanobacteria strains were isolated in Lake Iznik in northwestern Turkey. Morphological characterization of these strains suggested all three strains conformed to classical taxonomic identification of A. gracile with some differences such as clumping of filaments, partially hyaline cells in some filaments and longer than usual vegetative cells. Sequences of 16S rRNA gene of these strains were placed within an A. gracile cluster including the majority of PST producing strains, confirming the identification of these strains as A. gracile . These new strains possessed saxitoxin biosynthesis genes sxtA, sxtG and their sequences clustered with those of other A. gracile . Liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis demonstrated the presence of NEO, STX, dcSTX and decarbamoylneosaxitoxin (dcNEO) in all strains. This is the first report of a PST producer in any water body in Turkey and first observation of dcNEO in an A. gracile culture. [ABSTRACT FROM AUTHOR]

In October 2012, paralytic shellfish toxins (PST) were detected in the hepatopancreas of Southern Rock Lobsters ( Jasus edwardsii) collected from the east coast of Tasmania, Australia. This resulted in the first commercial closure in Australia for this species. Questions were raised on how the toxins were transferred to the lobsters, how long the toxins would persist, whether PST-contaminated hepatopancreas posed a risk to human health, and what management strategies could be applied. The aim of this study was to investigate whether PST-contaminated mussels are a potential vector enabling toxin accumulation in J. edwardsii and to collect information on toxin uptake, distribution and depuration rates and toxin profiles under controlled experimental settings. Lobsters were fed mussels naturally contaminated with PST for a period of 28 days in an experimental setting; following this, lobsters were allocated to either fed or starved treatment groups. PST were not detected in the tail tissue of lobsters at any stage of the experiment. Lobster hepatopancreas contained mean levels of 2.4 mg STX.2HCl eq/kg after 28 days of uptake, although substantial variability in total toxicity was observed. The PST profile of the hepatopancreas was similar to that of the contaminated mussels used as feed. Significant differences were noted in the PST depuration rates between fed and starved treatment groups. The daily depuration rate for total PST was estimated to be 0.019 and 0.013 mg STX.2HCl eq/kg for the fed and starved treatment groups respectively using a constant-rate decay model. After 42 days of depuration, total PST (STX equivalents) levels in the hepatopancreas of all lobsters were below 0.8 mg STX.2HCl eq/kg, which represents the regulatory level applied to bivalves. This result indicates that long-term holding to depurate PST may potentially be used as a risk management tool. [ABSTRACT FROM AUTHOR]

The cyanobacterium Dolichospermum circinale (formerly Anabaena circinalis ) is responsible for neurotoxic saxitoxin-producing blooms in Australia. Previous studies have reported distinct isolates of toxic D. circinale producing different saxitoxin analogues at varying amounts, but the mechanisms responsible remain poorly understood. To assess the characteristics that may be responsible for this variance, a morphological, molecular and chemical survey of 28 Anabaena isolates was conducted. Morphological characteristics, presence or absence of saxitoxin biosynthetic genes and toxin amount and profile were assessed. The 28 isolates were collected from 16 locations. A correlation between the size of the isolates and its reported toxicity or geographical location could not be found. Molecular screening for the presence of several sxt genes revealed eight out of the 28 strains harboured the sxt gene cluster and all tailoring genes except sxtX . Furthermore, the presence of PSTs was correlated with the presence of the sxt cluster using quantitative pre-column oxidation high performance liquid chromatography with fluorescence detection (HPLC-FLD) and LC-MS/MS. Interestingly, isolates differed in the amount and type of toxins produced, with the eight toxic strains containing the core and tailoring biosynthetic genes while non-toxic strains were devoid of these genes. Moreover, the presence of sxt tailoring genes in toxic strains correlated with the biosynthesis of analogues. A greater understanding of toxin profile/quantity from distinct sites around Australia will aid the management of these at-risk areas and provide information on the molecular control or physiological characteristics responsible for toxin production. [ABSTRACT FROM AUTHOR]

Trichodesmium is an enigmatic bloom forming, non-heterocystous cyanobacterium reported most frequently in the coastal waters of India. However, the toxigenic potential of this globally significant N 2 fixing cyanobacterium has not been characterized. In this study, we report for the first time the presence of potent multi-class neurotoxins such as Anatoxin-a, Saxitoxins, Gonyautoxin and hepatotoxins like MC-LR, MC-YA from a bloom material of Trichodesmium sp. MBDU 524 collected at the Gulf of Mannar region. Toxins were determined using liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS) analysis of HPLC purified aqueous and solvent fractions. Molecular phylogenetic analysis through 16S rRNA gene sequencing showed the close relationship with Trichodesmium erythraeum clade. The toxigenic potential was validated through brine shrimp toxicity assay and showed 100% mortality after 48 h of incubation. The results suggest the potential toxigenic and environmental impacts of Trichodesmium bloom sample from the Gulf of Mannar region. [ABSTRACT FROM AUTHOR]

Saxitoxin (STX) is a potent marine toxin that causes paralytic shellfish poisoning (PSP) which can result in significant morbidity and mortality in humans. Low lethal doses, rapid onset of PSP symptoms, and brief STX half-life in vivo require sensitive and rapid diagnostic techniques to monitor human exposures. Our laboratory has validated an enzyme-linked immunosorbent assay (ELISA) for quantitative detection of STX from 0.020 to 0.80 ng/mL in human whole blood and from 0.06 to 2.0 ng/mL in dried human blood which is simple, sensitive, rapid, and cost-effective. To our knowledge, this is the first validated method for the quantitation of saxitoxin in whole blood. Microsampling devices were used in sample collection which allows for standardized collection of blood, stable storage, and cost-efficient shipping. Quality control precision and accuracy were evaluated over the course of validation and were within 20% of theoretical concentrations. No detectable background concentrations of STX were found among fifty whole blood and dried blood convenience samples. Additionally, ten spiked individual whole blood and dried blood samples were tested for accuracy and precision and were within 20% of theoretical concentrations. Gonyautoxins 2&3 (GTX2&3) cross-reacted with this ELISA by 21%, but all other structurally related PSP toxins tested cross-reacted less than two percent. While clinical diagnosis or treatment of PSP would be unaffected by GTX2&3 cross-reactivity by ELISA, to accurately quantify individual PSP toxins, these results should be coupled with high performance liquid chromatography mass spectrometry measurements. [ABSTRACT FROM AUTHOR]

Last decades, cyanobacterial blooms have been commonly reported in Russia. Among the boom-forming species, potential toxin producers have been identified. The aim of this paper was to study the presence of neurotoxic compounds - saxitoxins and anatoxin-a - in water bodies from different regions of Russia. We also made attempts to identify the neurotoxin-producing genera. The good convergence of the results obtained by light microscopy, PCR and LC-MS/MS analyses indicated the presence of active neurotoxin producing species in all investigated water bodies. Saxitoxin was detected in phytoplankton from 4 water bodies in Central European Russia and West Siberia, including lake and reservoirs used as a source for potable water. The water bodies differed with the respect of saxitoxin producers which belonged to Aphanizomenon and/or Dolichospermum genera. For the first time, we obtained quantitative data on the intracellular saxitoxin concentration in Russian freshwaters using LC-MS/MS. Anatoxin-a was detected only in lakes of Northwestern Russia. In the eutrophic shallow Lower Suzdal Lake, Aphanizomenon was the stated anatoxin-a-producing genus. In the large shallow artificial hypertrophic Sestroretskij Razliv Lake, it was very likely that both dominant species - Aphanizomenon flos-aquae and Dolichospermum planctonicum - were anatoxin-a producers. [ABSTRACT FROM AUTHOR]

Diamondback terrapins ( Malaclemys terrapin ) are a threatened or endangered species in much of their range along the U.S. Atlantic and Gulf coasts. Over an approximately three-week period from late April to mid-May 2015, hundreds of adult diamondback terrapins were found dead on the shores of Flanders Bay, Long Island, New York, USA. Concurrent with the mortality event, elevated densities of the paralytic shellfish toxin (PST)-producing dinoflagellate, Alexandrium fundyense (>10 4 cells L −1 ) and high levels of PST in bivalves (maximal levels = 540 μg STX eq. 100 g −1 shellfish tissue) were observed in the Flanders Bay region, resulting in shellfish bed closures in regional tributaries. Gross and histologic postmortem examinations of terrapins revealed no physical trauma to individuals or a common, underlying disease process to explain the deaths. PST compounds (0.2–12.5 μg STX eq. 100 g −1 ) were present in various M. terrapin tissues collected over the duration of the mortality event. High-throughput sequencing revealed that the ribbed mussel ( Geukensia demissa , a PST vector) was present in the gastrointestinal tracks of all terrapin samples tested. While the potential of PST to cause mortality in chelonians has not been well-characterized, in the absence of other significant findings from necropsies and pathological analyses, we provide evidence that PST in shellfish was likely high enough to cause or contribute to the mortality in these small (<2.0 kg) animals. [ABSTRACT FROM AUTHOR]

The cyanobacterium Cylindrospermopsis raciborskii is an invasive species in water supply reservoirs worldwide, which can produces cylindrospermopsins and saxitoxins. In the wild, guppy ( Poecilia vivipara ) can be exposed to cyanotoxins, but those born and reared in laboratory are free of this contact. The aim of this paper was to comparatively measure the locomotor activity of ‘wild’ and ‘lab’ P . vivipara before and after exposure to crude extracts of two different cultures of C . raciborskii (CYRF-01), a saxitoxin-procucer strain. The movement of each fish was recorded using an image monitoring system (Videomex V ® ) before and after 48 h exposure to cyanobacterial extracts. Each experiment was performed during 4 h, with 1 h acclimation and 3 h recording period of the parameters Distance performed (DP), Swimming time (SwT), Stereotypic time (StT), Resting time (RT) and Average speed (AS). The quantification of saxitoxin in the solutions was performed by the enzyme-linked immunosorbent assay (ELISA). The weight or the total length did not influence the locomotor activity of fish in any of the experiments. The saxitoxin value was similar for both cultures (Culture 1: 7.3 μg L −1 and Culture 2: 8.6 μg L −1 ). However, in experiments with Culture 1 an increased activity in most parameters was observed, while in Culture 2, a decreased activity was observed only in ‘lab’ fish. Wild fish was less affected, showing higher resistance to both cyanobacterial crude extracts. This study showed that different cultures of the same strain of C . raciborskii and with similar contents of saxitoxin are able to change the locomotor activity of P . vivipara , contributing to the validation of the use of behavioral parameters to the evaluation of sublethal effects of toxic cyanobacteria on fish. [ABSTRACT FROM AUTHOR]

Cyanobacteria were screened from the surface of diseased sponges, stone and bedrock in Lake Baikal for the presence of saxitoxin using enzyme-linked immunosorbent assay. In sequel, eight paralytic shellfish toxin (PST) variants were identified using a MALDI mass spectrometry. Microscopic examination found that Tolypothrix distorta dominated in the biofouling samples. PCR and sequencing detected sxt A gene involved in saxitoxin biosynthesis, thereby providing evidence of the PST producing potential of Baikal cyanobacterial communities inhabiting different substrates. [ABSTRACT FROM AUTHOR]

Buccinidae whelk Neptunea varicifera (Dall), Cardiidae clam Serripes laperousii (Deshayes), and two unknown species of whelk and clam were collected from the Arctic Chukchi Sea and sub-Arctic Bering Sea in July 2014. In this study, the mollusk samples were analyzed by different liquid chromatography-tandem quadrupole mass spectrometry (LC-MS/MS) methods for multiple shellfish toxins, including okadaic acid (OA), pectenotoxin (PTX), yessotoxin (YTX), azaspiracid (AZA), cyclic imines (CI), and saxitoxin (STX) groups. PTX2 (≈2.0 μg kg −1 whole tissues) was detected exclusively in the clam S. laperousii collected from the Chukchi Sea. OA and dinophysistoxin-1 (DTX1) were restricted to mollusk samples collected from the Bering Sea, and OA was the dominant component of the whelk N. varicifera (63 μg kg −1 digestive gland) and an unknown species of whelk (6.8 μg kg −1 digestive gland). Spirolide-1 (SPX1) was confirmed in most samples except for the whelk N. varicifera collected from the Bering Sea. The highest content of SPX1 (≈18.5 μg kg −1 digestive gland) occurred in the whelk N. varicifera collected from the Chukchi Sea, along with the suspected presence of SPX-C, SPX-D and didesMe-SPX-C. YTX, as well as its derivatives 45-OH-YTX and 45,46,47-Trinor-YTX, were found in all samples, with the highest YTX content (66 μg kg −1 digestive gland) present in the whelk N. varicifera collected from the Chukchi Sea. Interestingly, STX and dcSTX were measured only in the whelk N. varicifera and unknown species of clam collected from the Chukchi Sea. No AZA-group toxins, gymnodimine (GYM), or pinnatoxin G were found in any samples analyzed. Results demonstrated that the mollusk samples were contaminated by multiple shellfish toxins in the Chukchi and Bering seas. This study highlights the need to monitor potentially toxic microalgae in the Arctic and sub-Arctic regions, as well as species of mollusk that may be included in future commercial or subsistence harvests. [ABSTRACT FROM AUTHOR]

In the current communication we describe an innovative method to purify saxitoxin (STX), a toxin presents in contaminated muscle of Mylitus chilensis extracted in the southern part of Chile, using a liquid chromatographic methodology based on ionic pairs. The STX was extracted using HCl and treated with ammonium sulfate following a treatment with trichloroacetic acid and hexane/diethyl ether (97/3). The samples were analyzed by a semi-preparative HPLC in order to collect pure fractions of STX and these fractions were eluted in solid-phase cationic interchange SCX extraction columns. The purified STX was stable and homogeneous and its identity was confirmed by LC–MS–MS, which demonstrated a high quality purification of STX, without presence of analogs such as neosaxitoxin (Neo) and decarbamoyl saxitoxin (dcSTX). The STX biological activity was analyzed in a bioassay in mice model and compared to the standard STX produced by the FDA and no significant differences were observed. [ABSTRACT FROM AUTHOR]

The cyanobacterium Scytonema cf. crispum produces a range of saxitoxins. Previous studies on other saxitoxin-producing cyanobacteria have shown that toxin production can vary throughout the growth cycle. Monitoring cyanotoxin-production in S. cf. crispum is challenging because it is metaphytic and has a very slow growth rate (ca. 6 months to reach stationary phase). In this study, a new method was developed to track growth and toxin production in S. cf. crispum . Samples were collected once a week for 131 days, and cell concentrations and saxitoxin quotas determined. Cells in the lag and exponential growth phases had significantly ( P < 0.05) higher saxitoxin quotas (162 ± 37 fg cell −1 and 139 ± 32 fg cell −1 , respectively) than the stationary phases (83 ± 19 fg cell −1 ). Extracellular saxitoxin concentrations were present at low concentrations (2–16 ng mL −1 of culture medium) throughout the experiment. The proportion of extracellular saxitoxin to total saxitoxin decreased throughout the experiment. New knowledge on growth and saxitoxin variability will assist in improving monitoring, risk assessment and management of this species. [ABSTRACT FROM AUTHOR]

Saxitoxin (STX), a member of the family of paralytic shellfish poisoning toxins, poses toxicological and ecotoxicological risks. To develop an analytical recognition element for STX, a DNA aptamer (APT STX1 ) was previously discovered via an iterative process known as Systematic Evolution of Ligands by Exponential Enrichment (SELEX) by Handy et al. Our study focused on generating an improved aptamer based on APT STX1 through rational site-directed mutation and truncation. In this study, we generated the aptamer, M-30f, with a 30-fold higher affinity for STX compared with APT STX1 . The Kd value for M-30f was 133 nM, which was calculated by Bio-Layer Interferometry. After optimization, we detected and compared the interaction of STX with aptamers (APT STX1 or M-30f) through several techniques (ELISA, cell bioassay, and mouse bioassay). Both aptamers' STX-binding ability was demonstrated in all three methods. Moreover, M-30f performs better than its parent sequence with higher suppressive activity against STX. As a molecular recognition element, M-30f has good prospects for practical application. [ABSTRACT FROM AUTHOR]

Development of novel analytical tools to detect marine biotoxins has been warranted in view of the apparent global pervasiveness of algal-derived shellfish poisoning, and the limitations of existing methods. Here, we describe the initial phase in the development and evaluation of a tyrosine-containing analog of μ-conotoxin (μ-CTX) GIIIA as an alternative to saxitoxin (STX) in a receptor binding assay (RBA) for paralytic shellfish poisons. The peptide analog was synthesized and characterized for structure and bioactivity. The major product of oxidation elicited paralytic symptoms in mice at a minimum dose of 1.31 mg kg −1 (i.p.). Mass spectrometry analysis of the bioactive peptide gave a molecular mass of 2637.52 Da that was close to the predicted value. Iodination via chloramine-T produced non-, mono- and di-iodinated peptides (respectively, NIP, MIP and DIP). Competition assays against 3 H-STX revealed higher Ki and EC50 (P < 0.0001, ANOVA) indicating reduced affinity for the receptor, and limited displacement of receptor-bound STX. However, subsequent use of MIP may extend the application of RBA to detect small changes in toxin levels owing to its likely enhanced displacement by STX. This may be useful in analyzing samples with toxicities near the regulatory limit, or in establishing baseline values in high risk environments. [ABSTRACT FROM AUTHOR]

Saxitoxin (STX) and neosaxitoxin (NEO) are potent neurotoxins that cause paralytic shellfish poisoning (PSP). PSP typically occurs through the ingestion of bivalve shellfish that have consumed toxin producing dinoflagellates. Due to initial presentation of symptoms being nonspecific, a clinical measurement is needed to confirm exposure to these toxins. Our group has developed an online solid phase extraction hydrophilic interaction liquid chromatography (HILIC) method for the analysis of STX and NEO in human urine with tandem mass spectrometry. A unique feature of this online method is the incorporation of a new synthetic 15 N 4 -STX labeled internal standard used for quantitation. Manual sample preparation time was reduced by approximately 70% for 98 urine samples as compared to a previously reported method. The lowest reportable limit for STX was improved from 5.0 ng/mL to 1.01 ng/mL and from 10.0 ng/mL to 2.62 ng/mL for NEO. Three analysts validated the method with 20 calibration curves total over 30 days with precision and accuracy within ±15% for all QCs. This new online method rapidly identifies STX and NEO exposure with improved sensitivity, which can facilitate the work of public health authorities to confirm the cases of PSP, complementing the many shellfish monitoring programs worldwide. [ABSTRACT FROM AUTHOR]

Over the past 15 years or so, several Scottish harbour seal ( Phoca vitulina ) populations have declined in abundance and several factors have been considered as possible causes, including toxins from harmful algae. Here we explore whether a link could be established between two groups of toxins, domoic acid (DA) and saxitoxins (STXs), and the decline in the harbour seal populations in Scotland. We document the first evidence that harbour seals are exposed to both DA and STXs from consuming contaminated fish. Both groups of toxins were found in urine and faeces sampled from live captured (n = 162) and stranded animals (n = 23) and in faecal samples collected from seal haul-out sites (n = 214) between 2008 and 2013. The proportion of positive samples and the toxins levels measured in the excreta were significantly higher in areas where harbour seal abundance is in decline. There is also evidence that DA has immunomodulatory effects in harbour seals, including lymphocytopenia and monocytosis. Scottish harbour seals are exposed to DA and STXs through contaminated prey at potentially lethal levels and with this evidence we suggest that exposure to these toxins are likely to be important factors driving the harbour seal decline in some regions of Scotland. [ABSTRACT FROM AUTHOR]

Dinoflagellates of the genus Alexandrium produce the neurotoxin saxitoxin (STX), responsible for paralytic shellfish poisoning (PSP) and accumulates in marine invertebrates. The recent identification of STX biosynthesis genes allowed us to investigate the expression of sxtA4 at different growth stages in Alexandrium catenella Group IV. We found no significant differences in expression of sxtA4 , despite significant differences in STX levels at different growth stages ( P < 0.023). Three reference genes were tested for normalisation: actin, cytochrome b ( cob ), and the large subunit ribosomal RNA (LSU rDNA). cob was most stably expressed but the combination of two reference genes, actin and cob , resulted in the best stability factor. Most genomic sequences of sxtA4 from A. catenella were in a clade that included sequences from Alexandrium fundyense Group I, however, one paralogue was not related to the others, suggesting recombination or lateral transfer. A comparison of the sxtA4 cDNA sequences with genomic DNA sequences indicated the possibility of transcript editing and the preferential transcription of certain genomic DNA loci. The results show that, in dinoflagellates, post-transcriptional mechanisms play a major role in the regulation of saxitoxin biosynthesis. [ABSTRACT FROM AUTHOR]

The effects of temperature on growth, cell toxicity, toxin content, and profile of paralytic shellfish toxins was determined in eight isolates of Gymnodinium catenatum from several localities along the Pacific Coast of Mexico. The isolates were cultivated in modified f/2 media with Se (10 −8 M), and a reduced concentration of Cu (10 −8 M), under a 12 h:12 h day–night cycle with an irradiance of 150 μE m −2 s −1 . Isolates were progressively adapted for three generations to each of the temperatures (16, 19, 22, 24, 27, 30, and 33 °C). The cultures were grown in 125 mL Erlenmeyer flasks with 60 mL of media and harvested by filtration in late exponential growth. Toxins were analyzed by HPLC with a post-column oxidation and fluorescent detection (FLD). G. catenatum isolates tolerate temperatures between 16 and 33 °C, with maximum growth rates of 0.32 and 0.39 div day −1 at 21 °C and 24 °C, respectively; maximum cell densities of 4700 and 5500 cells mL −1 were obtained at 27 and 21 °C, respectively. No effect of toxicity per cell with temperature was observed, varying between 10.10 and 28.19 pgSXTeq cell −1 . Ten saxitoxin analogues were detected in all isolates, observing changes in the toxin profile with temperature. C1/2 toxins decreased from 80% mol at 16 °C to 20% mol at 33 °C, B1/2 toxins increased from 19% mol at 16 °C to 42% mol at 33 °C, and decarbamoyl toxins were more abundant at 21 °C. These results show that G. catenatum isolates from different regions of the Pacific coast of Mexico have a similar response to temperature and that this parameter can modify growth rate, cell density, and toxin profile of the species, particularly the decarbamoyl and sulfocarbamoyl toxins. [ABSTRACT FROM AUTHOR]

This is the first report of a bloom of the toxic marine dinoflagellate Alexandrium minutum in the Azores Archipelago (Northeastern Atlantic Ocean). In September 2013, high cell concentrations (1.3 × 10 7 cells L −1 ) of this species were recorded in Santo Cristo coastal lagoon causing an orange-brown water discoloration, death of small pelagic fish and toxification of shellfish resources ( Ruditapes decussatus ). Levels of Paralytic Shellfish Poisoning toxins in clams exceeded 30 times the regulatory limit, and were associated with the intoxication of four people. [ABSTRACT FROM AUTHOR]

We investigated the ability of recombinant pufferfish saxitoxin and tetrodotoxin binding protein types 1 and 2 of Takifugu rubripes (rTrub.PSTBP1 and rTrub.PSTBP2) to bind to tetrodotoxin (TTX) and tributyltin. Both rTrub.PSTBPs bound to tributyltin in an ultrafiltration binding assay but lost this ability on heat denaturation. In contrast, only rTrub.PSTBP2 bound to TTX even heat denaturation. This result suggests that the amino acid sequence of PSTBP2 may be contributed for its affinity for TTX. [ABSTRACT FROM AUTHOR]

Abstract: The aim of this study was establish a protocol for isolation and primary culture of neurons from tropical freshwater fish species Hoplias malabaricus for assessment of the effects of neurotoxic substances as saxitoxins (STXs). Cells from brain of H. malabaricus were treated with different concentrations of trypsin, dispase and papain for tissue dissociation. Cells type was separated by cellular gradient and basic fibroblast growth factor (bFGF) supplement nutrition media were added. The dissociated cells were plated with medium and different STXs concentrations and the toxic cellular effects such as oxidative stress, neurotoxicity, and genotoxicity and apoptosis process were evaluated. Cultures treated with bFGF showed the greatest adherence, survival and cellular development. STXs increased specific activity of glutathione peroxidase and lipoperoxidation levels, were cytotoxic and genotoxic indicated by the comet assay. Although the STXs effects due the blockage of sodium channels is reported to be reversible, the time exposure and concentration of STXs suggested cellular injuries which can lead to neuropathology. The establishment of primary neuronal culture protocol enables new applications for neurotoxicological assessments. [Copyright &y& Elsevier]

Abstract: Ocean acidification is considered a major threat to marine ecosystems and may particularly affect primary producers. Here we investigated the impact of elevated pCO2 on paralytic shellfish poisoning toxin (PST) content and composition in two strains of Alexandrium tamarense, Alex5 and Alex2. Experiments were carried out as dilute batch to keep carbonate chemistry unaltered over time. We observed only minor changes with respect to growth and elemental composition in response to elevated pCO2. For both strains, the cellular PST content, and in particular the associated cellular toxicity, was lower in the high CO2 treatments. In addition, Alex5 showed a shift in its PST composition from a non-sulfated analogue towards less toxic sulfated analogues with increasing pCO2. Transcriptomic analyses suggest that the ability of A. tamarense to maintain cellular homeostasis is predominantly regulated on the post-translational level rather than on the transcriptomic level. Furthermore, genes associated to secondary metabolite and amino acid metabolism in Alex5 were down-regulated in the high CO2 treatment, which may explain the lower PST content. Elevated pCO2 also induced up-regulation of a putative sulfotransferase sxtN homologue and a substantial down-regulation of several sulfatases. Such changes in sulfur metabolism may explain the shift in PST composition towards more sulfated analogues. All in all, our results indicate that elevated pCO2 will have minor consequences for growth and elemental composition, but may potentially reduce the cellular toxicity of A. tamarense. [Copyright &y& Elsevier]

Abstract: Saxitoxin (STX) is the most potent paralytic shellfish poisoning toxin in crustaceans and molluscs, and is known to cause intoxication to humans and marine animals due to its neurotoxicity. However, the extent of its early developmental toxicity to marine species remains unknown. In this study, we examined the early developmental toxicity of STX using marine medaka (Oryzias melastigma) embryos as model. The medaka embryos were exposed to STX for four days, from the early blastula stage onwards, and this exposure period covered the main developmental stage of the central nervous system and somites. After exposure, the treated medaka eleutheroembryos at 15 day post fertilization exhibited abnormal growth with longer body length and relatively smaller yolk sac size. High cell proliferation, neuron development, and metabolism were confirmed using whole-mount immunostaining and two-dimensional electrophoresis. In summary, STX disturbed the normal growth of medaka embryos probably by affecting the metabolic rate in the exposed medaka embryos. [Copyright &y& Elsevier]

Abstract: Cyanobacteria in surface water are well known for their ability to form toxic blooms responsible for animal mortality and human poisoning. Accompanying major progress in science and technology, the state of knowledge of cyanotoxins has dramatically increased over the last two decades. The bibliometric approach applied in this study shows the evolution of research and identifies major gaps to be filled by future work. Although the publication rate has gradually increased from one hundred to three hundred articles per year since the 1990s, half of the literature available focuses on microcystins and another quarter on saxitoxins. Other cyanotoxins such as beta-N-methylamino-l-alanine or cylindrospermopsin remain vastly disregarded. Moreover, most of the publications deal with toxicity and ecology while other research areas, such as environmental and public health, require additional investigation. The analysis of the literature highlights the main journals for the communication of knowledge on cyanotoxins but also reveals that 90% of the research is originated from only ten countries. These countries are also those with the highest H-index and average number of citation per article. Nonetheless, the ranking of these countries is significantly altered when the amount of publications is normalized based on the population, the number of universities, the national gross domestic product or the government revenue. However, the lower amount of publications from Eastern Europe, Africa and South America could also reflect the lack of monitoring campaigns in these regions. This lack could potentially lead to the underestimation of the prevalence of toxic cyanobacterial blooms and the diversity of toxins worldwide. [Copyright &y& Elsevier]

Abstract: Saxitoxin and its derivatives, the paralytic shellfish toxins (PSTs), are known to be toxic to humans, and maximum permitted levels in seafood have been established by regulatory authorities in many countries. Until recently, the mouse bioassay was the reference method for determining the levels of these toxins in seafood, but this has now been superseded by chemical methods of analysis. The latter methods are able to determine the levels of many PSTs in shellfish, but for risk assessment an estimate of the relative toxicities of the individual components of the PST mixture is required. The relative toxicities are expressed as “Toxicity Equivalence Factors” (TEFs). At present, TEFs are based on relative specific activities in the mouse bioassay, rather than on acute toxicity determinations, as measured by median lethal doses (LD50s). In the present study, the median lethal doses of saxitoxin, neosaxitoxin, decarbamoyl saxitoxin and equilibrium mixtures of gonyautoxins 1&4 and gonyautoxins 2&3 have been determined by intraperitoneal injection, gavage and feeding. The results indicate that specific activities in the MBA do not consistently correlate with acute toxicities by any of the routes of administration, and TEFs, particularly for neosaxitoxin, require revision. [Copyright &y& Elsevier]

A study was conducted to assess the feasibility for the production of sterile, stable and homogenous shellfish reference materials containing known concentrations of paralytic shellfish poisoning (PSP) toxins. Pacific oysters were contaminated with toxins following mass culturing of toxic algae and shellfish feeding experiments. Live oysters were shucked and tissues homogenised, before measuring into multiple aliquots, with one batch subjected to gamma irradiation treatment and the other remaining untreated. The homogeneity of both batches of samples was assessed using a pre-column oxidation liquid chromatography with fluorescence detection (Pre-COX LC-FLD) method and shown to be within the limits of normal within-batch repeatability. A twelve-month stability experiment was conducted for both untreated and gamma irradiated batches, specifically examining the effects of long term storage at −20 °C, +4 °C and +40 °C. Results indicated mostly good stability of PSP toxins in both materials when stored frozen at −20 °C, but with the instability of GTX2&3 concentrations in the untreated tissues eliminated in the irradiated tissues. Analysis using a post-column oxidation (PCOX) LC-FLD method also showed epimerisation in both GTX1&4 and GTX2&3 epimeric pairs in untreated samples after only 6 months frozen storage. This issue was not present in the tissues irradiated before long term storage. Biological activity testing confirmed the absence of bacteria in the irradiated samples throughout the 12 month study period. With such results there was clear evidence for the potential of increasing the scale of the mass culturing and shellfish feeding for the production of large batches of tissue suitable for the preparation of a certified matrix reference material. Overall results demonstrated the feasibility for production of oyster reference materials for PSTs, with evidence for prolonged stability following gamma irradiation treatment and storage at −20 °C. [ABSTRACT FROM AUTHOR]

Abstract: Pufferfish saxitoxin and tetrodotoxin binding protein (PSTBP) was previously isolated from the plasma of the marine pufferfish, Takifugu pardalis. In this study, we investigated distribution pattern of PSTBP in intestine, liver, ovary, skin, and skeletal muscle of T. pardalis by immunohistochemical staining for the study of functions of this protein. In the skin, dermis around the tetrodotoxin secreting gland was positive, while this secreting gland itself was negative. In the ovary containing vitellogenic oocytes, ovarian wall and vitelline envelope were positive, while yolk and nucleus were negative. In the liver, hepatocytes with large fat droplets and capillaries were positive. In the intestine, the lamina propria mucosae were positive, while the mucosal epithelium was negative. In the skeletal muscle, only capillaries were positive. Furthermore, liver specific expression of PSTBP was confirmed by Northern blot analysis. Based on these results together with reported tetrodotoxin localization pattern in pufferfish, PSTBP was assumed to be a carrier protein to transfer tetrodotoxin among the tissues, especially liver, ovary, and skin. [Copyright &y& Elsevier]

Abstract: The bioaccumulation of saxitoxins (STX) in the trophic chain, mainly in freshwater, are not completely known. This work aimed to elucidate the effects of STX on Hoplias malabaricus through trophic bioassay. The fish were fed once every five days with Astyanax sp. before being subjected to an intraperitoneal inoculation with the lysate of Cylindrospermopsis raciborskii culture containing 97% STX and 3% by neosaxitoxin and gonyautoxin during 20 days. The animal''s liver was assessed using biomarkers as activities of superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), and glutathione peroxidase (GPx), and concentrations of reduced glutathione (GSH) and lipoperoxidation (LPO) and protein carbonylation (PCO). In the blood was analyzed the genotoxic and hematological parameters. The hepatosomatic index and the relative condition factor did not show a significant difference between the exposed and control groups. The values of mean corpuscular hemoglobin concentration and mean corpuscular hemoglobin increased in the STX group. The hepatic tissue from both groups exhibited a typical pattern that have been already described for most teleost fish. The results suggested the generation of reactive oxygen species, with increased activity of GPx and concentrations of LPO and GSH; whereas the specific activity of SOD decreased. However, no changes were observed in the CAT, PCO, and DNA damage. Although the STX effects are known as neurotoxic, this cyanotoxin caused liver biochemical alterations that can be considered ecologically relevant. [Copyright &y& Elsevier]

Abstract: In many respects tetrodotoxin (TTX) is the quintessential natural toxin. It is unequivocally toxic to mammals with LD50 values for mice in the range of 10 μg/kg (intraperitoneal), 16 μg/kg (subcutaneous), and 332 μg/kg (oral) (Kao, 1966). Its biothreat status is recognized by its listing as a “Select Agent” by the US Department of Health and Human Services which includes regulated agents “determined to have the potential to pose a severe threat to both human and animal health” (http://www.selectagents.gov/). It has a well-defined cellular target (i.e., NaV channels) and pharmacological mode of action (i.e., block of nerve and muscle action potentials), and it is an indispensable chemical tool in neuroscience. It is widely distributed in marine and terrestrial ecosystems where it plays a role in the chemical ecology of predator–prey relationships and drives evolutionary selection of TTX-resistance (Hanifin, 2010; Williams, 2010; Zimmer and Ferrer, 2007). Lastly, TTX has acquired a certain mystique in scientific lore attributable to many fascinating aspects of its natural history and molecular interactions as presented in selected summary below. Additional information may be found in other excellent reviews (Fozzard and Lipkind, 2010; Kao, 1966; Lee and Ruben, 2008; Narahashi, 2001, 2008). [Copyright &y& Elsevier]

Abstract: Saxitoxin (STX) is a low molecular weight neurotoxin mainly produced by certain marine dinoflagellates that, along with its family of similarly related paralytic shellfish toxins, may cause the potentially fatal intoxication known as paralytic shellfish poisoning. Illness and fatality rates are low due to the effective monitoring programs that determine when toxins exceed the established regulatory action level and effectuate shellfish harvesting closures accordingly. Such monitoring programs rely on the ability to rapidly screen large volumes of samples. Many of the screening assays currently available employ antibodies or live animals. This research focused on developing an analytical recognition element that would eliminate the challenges associated with the limited availability of antibodies and the use of animals. Here we report the discovery of a DNA aptamer that targets STX. Concentration-dependent and selective binding of the aptamer to STX was determined using a surface plasmon resonance sensor. Not only does this work represent the first reported aptamer to STX, but also the first aptamer to any marine biotoxin. A novel strategy of using a toxin–protein conjugate for DNA aptamer selection was successfully implemented to overcome the challenges associated with aptamer selection to small molecules. Taking advantage of such an approach could lead to increased diversity and accessibility of aptamers to low molecular weight toxins, which could then be incorporated as analytical recognition elements in diagnostic assays for foodborne toxin detection. The selected STX aptamer sequence is provided here, making it available to any investigator for use in assay development for the detection of STX. [Copyright &y& Elsevier]