Search

Nicotine activates nicotinic acetylcholine receptors (nAChRs), which are overexpressed in numerous cancer types, leading to signaling pathways that increase lung cancer invasiveness and resistance to chemotherapeutic agents. In this study, the effects of APS12-2, a synthetic analog of marine sponge toxin that acts as an antagonist of nAChRs, was investigated in vitro on A549 human lung adenocarcinoma cells and non-tumorigenic human lung epithelial BEAS-2B cells. In addition, gelatin nanoparticles (GNPs) loaded with APS12-2 (APS12-2-GNPs) were prepared and their effects were compared with those of free APS12-2. Nicotine reduced cytotoxicity, the formation of reactive oxygen species, and the formation of lipid droplets caused by cisplatin on A549 cells. The effects of nicotine on the decreased efficacy of cisplatin were reduced by APS12-2 and APS12-2-GNPs. APS12-2-GNPs showed a substantial advantage compared with free APS12-2; the cytotoxicity of APS12-2 on BEAS-2B cells was greatly reduced when APS12-2 was loaded in GNPs, whereas the cytotoxicity on A549 cells was only slightly reduced. Our results suggest that both APS12-2 and APS12-2-GNPs hold promise as supportive agents in the cisplatin-based chemotherapy of lung cancer. [ABSTRACT FROM AUTHOR]

Nicotine binds to nicotinic acetylcholine receptors (nAChRs) that are overexpressed in different cancer cells, promoting tumor growth and resistance to chemotherapy. In this study, we aimed to investigate the potential of APS7-2 and APS8-2, synthetic analogs of a marine sponge toxin, to inhibit nicotine-mediated effects on A549 human lung cancer cells. Our electrophysiological measurements confirmed that APS7-2 and APS8-2 act as α7 nAChR antagonists. APS8-2 showed no cytotoxicity in A549 cells, while APS7-2 showed concentration-dependent cytotoxicity in A549 cells. The different cytotoxic responses of APS7-2 and APS8-2 emphasize the importance of the chemical structure in determining their cytotoxicity on cancer cells. Nicotine-mediated effects include increased cell viability and proliferation, elevated intracellular calcium levels, and reduced cisplatin-induced cytotoxicity and reactive oxygen species production (ROS) in A549 cells. These effects of nicotine were effectively attenuated by APS8-2, whereas APS7-2 was less effective. Our results suggest that APS8-2 is a promising new therapeutic agent in the chemotherapy of lung cancer. [ABSTRACT FROM AUTHOR]

The French Society of Toxinology (SFET), which celebrated its 30th anniversary this year, organized its 29th annual Meeting (RT29), shared by 87 participants, on 30 November–1 December 2023. The RT29 main theme, "Toxins: From the Wild to the Lab", focused on research in the field of animal venoms and animal, bacterial, fungal, or plant toxins, from their discovery in nature to their study in the laboratory. The exploration of the functions of toxins, their structures, their molecular or cellular ligands, their mode of action, and their potential therapeutic applications were emphasized during oral communications and posters through three sessions, of which each was dedicated to a secondary theme. A fourth, "miscellaneous" session allowed participants to present recent out-of-theme works. The abstracts of nine invited and 15 selected lectures, those of 24 posters, and the names of the Best Oral Communication and Best Poster awardees, are presented in this report. [ABSTRACT FROM AUTHOR]

Nicotine activates nicotinic acetylcholine receptors (nAChRs), which are overexpressed in numerous cancer types, leading to signaling pathways that increase lung cancer invasiveness and resistance to chemotherapeutic agents. In this study, the effects of APS12-2, a synthetic analog of marine sponge toxin that acts as an antagonist of nAChRs, was investigated in vitro on A549 human lung adenocarcinoma cells and non-tumorigenic human lung epithelial BEAS-2B cells. In addition, gelatin nanoparticles (GNPs) loaded with APS12-2 (APS12-2-GNPs) were prepared and their effects were compared with those of free APS12-2. Nicotine reduced cytotoxicity, the formation of reactive oxygen species, and the formation of lipid droplets caused by cisplatin on A549 cells. The effects of nicotine on the decreased efficacy of cisplatin were reduced by APS12-2 and APS12-2-GNPs. APS12-2-GNPs showed a substantial advantage compared with free APS12-2; the cytotoxicity of APS12-2 on BEAS-2B cells was greatly reduced when APS12-2 was loaded in GNPs, whereas the cytotoxicity on A549 cells was only slightly reduced. Our results suggest that both APS12-2 and APS12-2-GNPs hold promise as supportive agents in the cisplatin-based chemotherapy of lung cancer.

The discovery of a pufferfish specimen (Tetraodontidae) inside a frozen cuttlefish, purchased by a fishmonger, and caught in the Eastern Central Atlantic (FAO 34) is reported. The consumer, who reported this case to FishLab (Department of Veterinary Sciences, University of Pisa) for investigation, was a student of Veterinary Medicine at the University of Pisa. He recognized the Tetraodontidae because he attended practical lessons on fish morphological identification during the course of food inspection and was aware of the risks to human health linked to the Tetrodotoxin (TTX). In this study, the pufferfish was identified morphologically, using the FAO morphological keys, and molecularly, analyzing two markers, the cytochrome oxidase I (COI) and the cytochrome b genes, by DNA barcoding. The pufferfish was identified morphologically as Sphoeroides spp., and molecularly as Sphoeroides marmoratus using the COI gene (99-100% identity values). Literature reports that S. marmoratus from the Eastern Atlantic contains high concentrations of TTX in the gonads and the digestive tract. However, the possible passage of TTX from fish to other organisms linked to contact or ingestion has never been reported. This represents the first case of a potentially toxic pufferfish entering the market inside another organism. The fact that a student observed this occurrence highlights the key role of citizen science in the management of emerging risks. [ABSTRACT FROM AUTHOR]

Nicotine binds to nicotinic acetylcholine receptors (nAChRs) that are overexpressed in different cancer cells, promoting tumor growth and resistance to chemotherapy. In this study, we aimed to investigate the potential of APS7-2 and APS8-2, synthetic analogs of a marine sponge toxin, to inhibit nicotine-mediated effects on A549 human lung cancer cells. Our electrophysiological measurements confirmed that APS7-2 and APS8-2 act as α7 nAChR antagonists. APS8-2 showed no cytotoxicity in A549 cells, while APS7-2 showed concentration-dependent cytotoxicity in A549 cells. The different cytotoxic responses of APS7-2 and APS8-2 emphasize the importance of the chemical structure in determining their cytotoxicity on cancer cells. Nicotine-mediated effects include increased cell viability and proliferation, elevated intracellular calcium levels, and reduced cisplatin-induced cytotoxicity and reactive oxygen species production (ROS) in A549 cells. These effects of nicotine were effectively attenuated by APS8-2, whereas APS7-2 was less effective. Our results suggest that APS8-2 is a promising new therapeutic agent in the chemotherapy of lung cancer.

The French Society of Toxinology (SFET), which celebrated its 30th anniversary this year, organized its 29th annual Meeting (RT29), shared by 87 participants, on 30 November–1 December 2023. The RT29 main theme, “Toxins: From the Wild to the Lab”, focused on research in the field of animal venoms and animal, bacterial, fungal, or plant toxins, from their discovery in nature to their study in the laboratory. The exploration of the functions of toxins, their structures, their molecular or cellular ligands, their mode of action, and their potential therapeutic applications were emphasized during oral communications and posters through three sessions, of which each was dedicated to a secondary theme. A fourth, “miscellaneous” session allowed participants to present recent out-of-theme works. The abstracts of nine invited and 15 selected lectures, those of 24 posters, and the names of the Best Oral Communication and Best Poster awardees, are presented in this report.

The discovery of a pufferfish specimen (Tetraodontidae) inside a frozen cuttlefish, purchased by a fishmonger, and caught in the central-east Atlantic (FAO 34) is reported. The consumer, who reported this case to FishLab (Department of Veterinary Sciences, University of Pisa) for investigation, was a student of Veterinary Medicine at the University of Pisa. He recognized the Tetraodontidae because he attended practical lessons on fish morphological identification during the course of food inspection and was aware of the risks to human health linked to the Tetrodotoxin (TTX). In this study, the pufferfish was identified morphologically, using the FAO morphological keys, and molecularly, analyzing two markers, the cytochrome oxidase I (COI) and the cytochrome b genes, by DNA barcoding. The pufferfish was identified morphologically as Sphoeroides spp., and molecularly as Sphoeroides marmoratus using the COI gene (99-100% identity values). Literature reports that S. marmoratus from the eastern Atlantic contains high concentrations of TTX in the gonads and the digestive tract. However, the possible passage of TTX from fish to other organisms linked to contact or ingestion has never been reported. This represents the first case of a potentially toxic pufferfish entering the market inside another organism. The fact that a student observed this occurrence highlights the key role of citizen science in the management of emerging risks.

C17-sphinganine analog mycotoxin (C17-SAMT) has been characterized as the contaminant responsible for the atypical toxicity reported in mussels from the Bizerte lagoon (northern Tunisia) over the past decade. C17-SAMT exhibited common symptoms of toxicity in mice, including flaccid paralysis and severe respiratory distress, followed by rapid death. To determine the potential health risks of this neurotoxin, we assessed its subchronic toxicity according to the recommendations of OCDE n° 407. The body weight and the structural changes of vital organs were recorded. Biochemical and hematological parameters were also quantified. Macroscopic observations showed that mice treated with 0.9, 9, and 90 µg/kg C17-SAMT had significantly reduced stomach weights, swollen and fragile intestines, and signs of nephritis with renal abscesses. Transaminase assays pointed out that exposure to C17-SAMT can lead to transaminitis. Above-average lactate dehydrogenase values were recorded in both the treated and satellite groups. Hematology data showed a significant reduction in red blood cell counts in high-dose-treated group. Reductions in hemoglobin and hematocrit were also recorded. Mean leukocyte counts were significantly elevated in the high-, mid-dose treated and satellite groups. At the microscopic level, we noted myocardial atrophy and hyperemia. In the lungs, we noted necrosis associated with macrophages perivascular infiltration and congestion. The kidneys showed mild inflammation and glomerular atrophy. The stomach exhibited mucosal atrophy, while a thin colon and distended small intestine were observed in high-dose-treated group. The liver was affected by vascular congestion, inflammatory infiltration, and lobular necrosis that evolved into acute hepatitis. Lesions, such as inflammatory infiltration and mild necrosis of the liver, cortical abscess with central necrosis in the kidney, and mild congestion of cardiac tissue were recorded in the satellite group.

The French Society of Toxinology (SFET) organized its 28th annual meeting on 28–29 November 2022 (RT28). The central theme of this meeting was "Toxins: What's up, Doc?", emphasizing the latest findings on animal, bacterial, algal, plant and fungal toxins through sessions dedicated to deep learning, toxin tracking and toxinomic advances, shared by ca. 80 participants. The abstracts of the 10 invited and 11 selected lectures and 15 posters, along with the names of the Best Oral Communication and Best Poster awardees, are presented in this report. [ABSTRACT FROM AUTHOR]

The toxic flatworm, Planocera multitentaculata, possesses highly concentrated tetrodotoxin (TTX), also known as pufferfish toxin, throughout its life cycle, including the egg and larval stages. Additionally, TTX analogues, 5,6,11-trideoxyTTX and 11-norTTX-6(S)-ol, have also been detected in the flatworm. The high concentration of TTX in the eggs and larvae appears to be for protection against predation, and 11-norTTX-6(S)-ol in the pharyngeal tissue in the adults is likely used to sedate or kill prey during predation. However, information on the role of 5,6,11-trideoxyTTX, a potential important biosynthetic intermediate of TTX, in the toxic flatworm is lacking. Here, we aimed to determine the region of localization of TTX and its analogues in the flatworm body, understand their pharmacokinetics during maturation, and speculate on their function. Flatworm specimens in four stages of maturity, namely juvenile, mating, spawning, and late spawning, were subjected to LC–MS/MS analysis, using the pharyngeal tissue, oocytes in seminal receptacle, sperm, and tissue from 12 other sites. Although TTX was consistently high in the pharyngeal tissue throughout maturation, it was extremely high in the oocytes during the spawning period. Meanwhile, 5,6,11-trideoxyTTX was almost undetectable in the pharyngeal part throughout the maturation but was very abundant in the oocytes during spawning. 11-norTTX-6(S)-ol consistently localized in the pharyngeal tissue. Although the localization of TTX and its analogues was approximately consistent with the MS imaging data, TTX and 11-norTTX-6(S)-ol were found to be highly localized in the parenchyma surrounding the pharynx, which suggests the parenchyma is involved in the accumulation and production of TTXs. [ABSTRACT FROM AUTHOR]

Tetrodotoxin (TTX) is a marine biotoxin whose biosynthesis is associated with the pufferfish. Its distribution is primarily focused in Asian and tropical marine areas. Currently, this group of toxins is classified as emerging in Europe, and its presence could be related to climate change. This incidence has prompted the European Union, with the European Food Safety Authority, to establish control and monitoring mechanisms for TTX in marine products in Europe. In this context, the development of analytical tools capable of ensuring the safety of food products, especially seafood and fish, is a crucial task. This study describes the development of a molecularly imprinted polymer (MIP) based electrochemical sensor for the analysis of TTX. The MIP was synthesized through the electropolymerization of a functional monomer, ortho -phenylenediamine in the presence of a dummy template, voglibose. The MIP sensor was constructed on a screen-printed gold electrode and characterized by cyclic voltammetry. Differential pulse voltammetry, using a redox probe ([Fe(CN) 6 ]3-/4-), was used in the analysis protocol. The developed sensor exhibited a linear response between 5.0 μg mL−1 and 25.0 μg mL−1, with a limit of detection of 1.14 μg mL−1. Its high imprinting efficiency conferred outstanding selectivity towards TTX. The sensor's applicability was confirmed through recovery assays on spiked mussel samples, achieving recoveries of 81.0 %, 110.2 %, and 102.5 % for external standard addition at 30.0, 44.0, and 60.0 μg kg−1, respectively, with relative standard deviations below 15 %. These results are comparable to those obtained using Hydrophilic Interaction Liquid Chromatography coupled with Tandem Mass Spectrometry, a validated method carried out by the European Reference Laboratory for Marine Biotoxins. Thus, the MIP sensor represents a portable, simple, and fast tool with essential analytical functionalities for the sampling phase and pre-selection of laboratory samples for analysis. [Display omitted] • Analysing tetrodotoxin can avoid severe or even fatal poisoning. • A simple and selective electrochemical MIP sensor was developed for tetrodotoxin. • A linear range from 5.0 to 25.0 μg mL−1 and an LOD of 1.14 μg mL−1 were achieved. • The sensor will allow significant time and cost savings in tetrodotoxin analysis. [ABSTRACT FROM AUTHOR]

Addressing the critical health concerns posed by domoic acid (DA), a neurotoxic compound produced by toxic marine algae and bioaccumulated in shellfish, necessitates the development of a rapid, precise, and robust detection system. Traditional DA detection methods have stability and sensitivity issues, which hinder effective toxin detection. To overcome these limitations, we developed a novel direct competitive enzyme-linked immunosorbent assay (dc-ELISA) platform that utilizes peptide-immobilized magnetic beads (MGBs/peptide). The affinity peptides identified through phage display and chemically synthesized with biotin labels present an innovative alternative to conventional antibodies for ELISA applications. Streptavidin-modified MGBs were used as the bioreceptor carriers to facilitate magnetic separation and simplify sample preparation, making the MGB/peptide-based dc-ELISA platform an ideal tool for comprehensive monitoring efforts. The developed platform exhibits a detection range of 0.5–10 ng mL−1 and a low limit of detection of 0.29 ng mL−1, offering enhanced sensitivity and cost-effectiveness. Moreover, our developed dc-ELISA demonstrated a high recovery rate when validated with DA-spiked CRM-mussel samples. This method overcomes the limitations of traditional detection techniques and offers a scalable and efficient approach to marine toxin surveillance with improved marine environmental monitoring and public health management. [Display omitted] • Domoic acid-specific peptide was identified, and its binding site was verified via molecular docking simulations. • Affinity peptide-immobilized magnetic bead-based direct competitive (dc)-ELISA for domoic acid detection was developed. • Proposed peptide-based dc-ELISA performs better than commercial ELISA kits. • Developed dc-ELISA can accurately detect domoic acid in real shellfish samples. [ABSTRACT FROM AUTHOR]

Saxitoxin (STX), which is produced by certain dinoflagellate species, is a type of paralytic shellfish poisoning toxin that poses a serious threat to human health and the environment. Therefore, developing a technology for the convenient and cost-effective detection of STX is imperative. In this study, we developed an affinity peptide-imprinted polymer-based indirect competitive ELISA (ic-ELISA) without using enzyme–toxin conjugates. AuNP/Co 3 O 4 @Mg/Al cLDH was synthesized by calcining AuNP/ZIF-67@Mg/Al LDH, which was obtained by combining AuNPs, ZIF-67, and flower-like Mg/Al LDH. This synthesized nanozyme exhibited high catalytic activity (K m = 0.24 mM for TMB and 132.5 mM for H 2 O 2). The affinity peptide-imprinted polymer (MIP) was imprinted with an STX-specific template peptide (STX MIP) on a multi-well microplate and then reacted with an STX-specific signal peptide (STX SP). The interaction between the STX SP and MIP was detected using a streptavidin-coated nanozyme (SA-AuNP/Co 3 O 4 @Mg/Al cLDH). The developed MIP-based ic-ELISA exhibited excellent selectivity and sensitivity, with a limit of detection of 3.17 ng/mL (equivalent: 0.317 μg/g). Furthermore, the system was validated using a commercial ELISA kit and mussel tissue samples, and it demonstrated a high STX recovery with a low coefficient of variation. These results imply that the developed ic-ELISA can be used to detect STX in real samples. [ABSTRACT FROM AUTHOR]

Okadaic acid (OA), which is one of the representative diarrhetic shellfish poisonings, is a prevalent food-poisoning toxin in the world. As OA consumption induces various digestive diseases, a strict monitoring tool for OA is highly required. In this study, we demonstrate the development of an aptamer-based fluorometric OA sensor. First, the affinity of the aptamer towards OA was confirmed through electrochemical impedance spectroscopy, and it was then utilized to develop the fluorometric sensor. The verified OA-specific aptamer was modified with 5-carboxyfluorescein (FAM) as a fluorophore, and graphene oxide (GO) was adopted as the quencher. By virtue of the quenching phenomena between FAM and GO, the developed sensor revealed a low detection limit of 6.35 ppb. Also, this sensor represented a high response to OA compared to other types of marine toxins, such as saxitoxin, domoic acid, and tetrodotoxin. [ABSTRACT FROM AUTHOR]

The French Society of Toxinology (SFET) organized its 28th annual meeting on 28–29 November 2022 (RT28). The central theme of this meeting was “Toxins: What’s up, Doc?”, emphasizing the latest findings on animal, bacterial, algal, plant and fungal toxins through sessions dedicated to deep learning, toxin tracking and toxinomic advances, shared by ca. 80 participants. The abstracts of the 10 invited and 11 selected lectures and 15 posters, along with the names of the Best Oral Communication and Best Poster awardees, are presented in this report.

Multiple animal species have evolved resistance to the neurotoxin tetrodotoxin (TTX) through changes in voltage-gated sodium ion channels (VGSCs). Amino acid substitutions in TTX-resistant lineages appear to be positionally convergent with changes in homologous residues associated with reductions in TTX block. We used homology modeling coupled with docking simulations to test whether positionally convergent substitutions generate functional convergence at the level of TTX–channel interactions. We found little evidence that amino acids at convergent positions generated similar patterns among TTX-resistant animal lineages across several metrics, including number of polar contacts, polar contact position, and estimates of binding energy. Though binding energy values calculated for TTX docking were reduced for some TTX-resistant channels, not all TTX-resistant channels and not all of our analyses returned reduced binding energy values for TTX-resistant channels. Our results do not support a simple model of toxin resistance where a reduced number of bonds between TTX and the channel protein prevents blocking. Rather models that incorporate flexibility and movement of the protein overall may better describe how homologous substitutions in the channel cause changes in TTX block.

The French Society of Toxinology (SFET) organized its 27th annual meeting on 9–10 December 2021 as a virtual meeting (e-RT27). The central theme of this meeting was “Toxins: Mr Hyde or Dr Jekyll?”, emphasizing the latest findings on plant, fungal, algal, animal and bacterial toxins during 10 lectures, 15 oral communications (shorter lectures) and 20 posters shared by ca. 80 participants. The abstracts of lectures and posters, as well as the winners of the best oral communication and poster awards, are presented in this report.

Throughout history, humans have encountered natural toxic chemicals from the ocean environment, often through contaminated seafood. Although marine toxins can be harmful to human health and devastate local environments when they are produced during algal bloom events, they are also important biochemical research reagents and drug leads in medicine. In spite of their long history, the biosynthetic origin of many well-known marine toxins has remained elusive. New biosynthetic insights have shed light on the chemical transformations that create the complex structures of several iconic oceanic toxins. To that end, this review highlights advances made in the biosynthetic understanding of five important environmental toxins of marine origin: domoic acid, kainic acid, saxitoxin, tetrodotoxin, and polyether polyketides such as brevetoxin. [ABSTRACT FROM AUTHOR]

Gold nanostars (GNST), gold nanospheres (GNP) and carbon black (CB) are chosen as alternative nanomaterials to modify carbon screen-printed electrodes (c-SPEs). The resulting three kinds of modified c-SPEs (GNP-SPE, CB-SPE and GNSP-SPE) were electrochemically and microscopically characterized and compared with standardized c-SPEs after pretreatment with phosphate buffer by pre-anodization (pre-SPE). The results show outstanding electrochemical performance of the carbon black-modified SPEs which show low transient current, low capacitance and good porosity. A competitive chronoamperometric immunoassay for the shellfish toxin domoic acid (DA) is described. The performances of the CB-SPE, GNP-SPE and pre-SPE were compared. Hapten-functionalized magnetic beads were used to avoid individual c-SPE functionalization with antibody while enhancing the signal by creating optimum surface proximity for electron transfer reactions. This comparison shows that the CB-SPE biosensor operated best at a potential near − 50 mV (vs. Ag/AgCl) and enables DA to be determined with a detection limit that is tenfold lower compared to pre-SPE (4 vs. 0.4 ng mL−1). These results show very good agreement with HPLC data when analysing contaminated scallops, and the LOD is 0.7 mg DA kg−1 of shellfish. [ABSTRACT FROM AUTHOR]

Food safety of aquatic products has attracted considerable attention worldwide. Although a series of conventional bioassays and instrumental methods have been developed for the detection of pathogenic bacteria, heavy metal residues, marine toxins, and biogenic amines during the production and storage of fish, shrimp, crabs et al., the nanotechnology-based analyses still have their advantages and are promising since they are cost-efficient, highly sensitive and selective, easy to conduct, facial design, often require no sophisticated instruments but with excellent detection performance. This review aims to summarize the advances of various biosensing strategies for bacteria, metal ions, and small molecule contaminants in aquatic products during the last five years, The review highlights the development in nanotechnologies applied for biorecognition process, signal transduction and amplification methods in each novel approach, the nuclease-mediated DNA amplification, nanomaterials (noble metal nanoparticle, metal-organic frameworks, carbon dots), lateral flow-based biosensor, surface-enhanced Raman scattering, microfluidic chip, and molecular imprinting technologies were especially emphasized. Moreover, this study provides a view of current accomplishments, challenges, and future development directions of nanotechnology in aquatic product safety evaluation. [Display omitted] • Nanotechnology used for aquatic products safety evaluation is reviewed. • Nanotechnology used for pathogens, metal ions, biotoxins and biogenic amines detection are reviewed. • The advantages of nanotechnology compared with traditional methods are discussed. • The shortage and future development of nanotechnology are provided. [ABSTRACT FROM AUTHOR]

A puffer fish poisoning case was reported from the coastal city of Veraval in the Gujarat state of India with patient reporting symptoms of giddiness, vertigo, aphasia and heaviness of head following consumption of cooked fish. Treatment was purely symptomatic and supportive. The patient was discharged from the hospital in a stable condition after 4 days. The suspected fish species was later identified using DNA (Deoxyribonucleic Acid) sequencing as Arothron stellatus with 100% identity. Arothron stellatus, also known as Stellate Puffer, is a demersal marine fish usually found in the Indian Ocean. [Display omitted] • There is lack of official data regarding puffer fish poisoning in India. • A puffer fish poisoning incident occurred in the coastal city of Veraval in India. • Treatments were symptomatic and supportive for the patient. • The fish was later identified as Stellate Puffer, Arothron stellatus. [ABSTRACT FROM AUTHOR]

Most marine biotoxins are produced by microalgae. The neurotoxin tetrodotoxin (TTX) has been reported in many seafood species worldwide but its source is unknown, making accumulation and depuration studies in shellfish difficult. Tetrodotoxin is a water-soluble toxin and cannot be directly ingested by shellfish. In the present study, a method was developed which involved binding TTX to solid particles of humic acid and encapsulating them in agar-gelatin capsules. A controlled quantity of TTX-containing microcapsules (size range 20–280 μm) was fed to Paphies australis, a bivalve known to accumulate TTX in the wild. The TTX-containing microcapsules were fed to P. australis every second day for 13 days. Ten P. australis (including five controls fed non-toxic microalgae) were harvested after 7 days and ten after 13 days. Paphies australis accumulated TTX, reaching concentrations of up to 103 µg kg−1 by day 13, exceeding the European Food Safety Authority recommended concentration of 44 μg kg−1 in shellfish. This novel method will allow future studies to explore the effects, accumulation and depuration rates of TTX in different animals and document how it is transferred through food webs.

[Display omitted] • A sensitive and novel mDAIA-NFF sensor was first proposed for OA detection. • A mismatched duplexed aptamer based on conformational change for OA was designed. • A label-free RCA coupled with ThT can further enhance the detection sensitivity. • This sensor detected OA with a LOD of 3.1 ng/L and showed a good linear range. • The method is compatible with handheld fluorimeters for further great promise. We developed a highly sensitive fluorescent assay to detect okadaic acid (OA), a prevalent aquatic toxin posing serious health risks. Our approach uses a mismatched duplexed aptamer (DA) immobilized on streptavidin-conjugated magnetic beads (SMBs) to create a DA@SMB complex. In the presence of OA, the cDNA unwinds, hybridizes with a G-rich segment pre-encoding circular template (CT), and undergoes rolling circle amplification (RCA) to produce G-quadruplexes, which are detected using the fluorescent dye thioflavine T (ThT). The method has a LOD of 3.1 × 10-3 ng/mL, a linear range of 0.1 ∼ 1.0 × 103 ng/mL, and was successfully applied to shellfish samples with spiked recoveries of 85.9% ∼ 102.2% and RSD less than 13%. Furthermore, instrumental analysis confirmed the accuracy and reliability of this rapid detection method. Overall, this work represents a significant advancement in the field of rapid aquatic toxin detection and has important implications for public health and safety. [ABSTRACT FROM AUTHOR]

[Display omitted] • Peptide-based competitive ELISA for detection of okadaic acid was developed. • Okadaic acid-specific peptide was successfully identified and chemically synthesized. • Developed peptide-based ELISA exhibit good limit of detection and limit of quantitation. • Our developed ELISA show good performance in a complexed real shellfish samples. Okadaic acid (OA) is a type of marine biotoxin produced by some species of dinoflagellates in marine environments. Consumption of shellfish contaminated with OA can cause diarrhetic shellfish poisoning (DSP) in humans with symptoms that typically include abdominal pain, diarrhea and vomiting. In this study, we developed an affinity peptide-based direct competition enzyme-linked immunosorbent assay (dc-ELISA) for the detection of OA in real samples. The OA-specific peptide was successfully identified via M13 biopanning and a series of peptides were chemically synthesized and characterized their recognition activities. The dc-ELISA system showed good sensitivity and selectivity with a half-maximal inhibitory concentration (IC 50) of 148.7 ng/mL and a limit of detection (LOD) of 5.41 ng/mL (equivalent, 21.52 ng/g). Moreover, the effectiveness of the developed dc-ELISA was validated using OA-spiked shellfish samples, and the developed dc-ELISA showed a high recovery rate. These results suggest that the affinity peptide-based dc-ELISA can be a promising tool for detecting OA in shellfish samples. [ABSTRACT FROM AUTHOR]

Chondrosia reniformis is a common marine demosponge showing many peculiarities, lacking silica spicules and with a body entirely formed by a dense collagenous matrix. In this paper, we have described the identification of a new cytotoxic protein (chondrosin) with selective activity against specific tumor cell lines, from C. reniformis, collected from the Liguria Sea. Chondrosin was extracted and purified using a salting out approach and molecular weight size exclusion chromatography. The cytotoxic fractions were then characterized by two-dimensional gel electrophoresis and mass spectrometry analysis and matched the results with C. reniformis transcriptome database. The procedure allowed for identifying a full-length cDNA encoding for a 199-amino acids (aa) polypeptide, with a signal peptide of 21 amino acids. The mature protein has a theoretical molecular weight of 19611.12 and an IP of 5.11. Cell toxicity assays showed a selective action against some tumor cell lines (RAW 264.7 murine leukemia cells in particular). Cell death was determined by extracellular calcium intake, followed by cytoplasmic reactive oxygen species overproduction. The in silico modelling of chondrosin showed a high structural homology with the N-terminal region of the ryanodine receptor/channel and a short identity with defensin. The results are discussed suggesting a possible specific interaction of chondrosin with the Cav 1.3 ion voltage calcium channel expressed on the target cell membranes.

Palytoxin has been found in several soft coral species which are popular for in-home reef aquariums. Although a limited number of case reports describing acute respiratory distress after inhalational exposure to palytoxin have been reported, there have been no reports regarding the potential long-term effects of inhalational exposure to palytoxin in the literature. This case follows an aquatic specialist in the U.S. over a period of seven years after a single intense occupational exposure to the aerosolized toxin from cleaning of a residential aquarium. • Occupational exposure to palytoxin from home aquarium maintenance. • Inhalational exposure to presumed aerosols generated during the handling of zoanthids. • Long-term cognitive and respiratory impairments from one-time exposure to palytoxin. [ABSTRACT FROM AUTHOR]

Pectenotoxin-2 (PCTX-2) is one of the polyether macrolide toxins isolated from scallops involved in diarrheic shellfish poisoning via actin depolymerization. In the present study, we examined the bioactive mechanism of PCTX-2 in smooth muscle cells and clarify mode of action of the PCTX-2-induced actin depolymerization using purified skeletal actin. PCTX-2 (300 nM-3 μM) non-selectively inhibited vascular smooth muscle contractions elicited by high K+ or phenylephrine in a dose-dependent manner. However, elevated cytosolic Ca2+ and myosin light chain phosphorylation stimulated by high K+ were only slightly inhibited by PCTX-2. By monitoring the fluorescent intensity of pyrenyl-actin, PCTX-2 was found to inhibit both the velocity and degree of actin polymerization. The critical concentration of G-actin was linearly increased in accordance with the concentration of PCTX-2, indicating sequestration of G-actin with 1 to 1 ratio. The kinetics of F-actin depolymerization by dilution assay indicated that PCTX-2 does not sever F-actin. Transmission electron microscopic and confocal microscopic observations demonstrated that PCTX-2 selectively depolymerized filamentous actin without affecting tublin. In conclusion, PCTX-2 is a potent natural actin depolymerizer which sequesters G-actin without severing F-actin. [ABSTRACT FROM AUTHOR]

Neosaxitoxin (NeoSTX) is a specific reversible blocker of voltage gated sodium channels on excitable cells. In the last decade, it has been tested in a number of interesting clinical trials, however there is still little information available on mammalian toxicity. Rats were treated for 12 weeks with doses of 1, 3 or 6 μg/kg of subcutaneous NeoSTX. At weeks 12 and 17, animals were sacrificed and blood samples collected for hematological and biochemical analysis. Organs were harvested for weight determination and histopathological assessments. The lowest acute toxicity via the intraperitoneal (ip) route was (30.35 μg/kg) and there was no significant difference between intramuscular and subcutaneous routes (11.4 and 12.41 μg/kg). The NeoSTX adiministration did not produce lethality at week 12 and after five weeks of suspension. NeoSTX 6 μg/kg ip produced reductions (p < 0.05) in body weight and food intake, and increased blood level of total and direct bilirubin, GGT and SGOT at week 12; all of these were reversed in the recovery period. NeoSTX 1 and 3 μg/kg ip did not show significant changes with the control group. Histopathological presentations were normal in all groups. This study revealed that NeoSTX is safe in vivo, giving a reliable security margin for its use like a local anesthetic.

Marine organisms are an increasingly important source of novel metabolites, some of which have already inspired or become new drugs. In addition, many of these molecules show a high degree of novelty from a structural and/or pharmacological point of view. Structure determination is generally achieved by the use of a variety of spectroscopic methods, among which NMR (nuclear magnetic resonance) plays a major role and determination of the stereochemical relationships within every new molecule is generally the most challenging part in structural determination. In this communication, we have chosen okadaic acid as a model compound to perform a computational chemistry study to predict 1H and 13C NMR chemical shifts. The effect of two different solvents and conformation on the ability of DFT (density functional theory) calculations to predict the correct stereoisomer has been studied.

We investigated an epidemic mortality cluster of yellow-eyed penguins (Megadyptes antipodes) that involved 67 moribund or dead birds found on various beaches of the Otago Peninsula, New Zealand, between 21 January and 20 March 2013. Twenty-four carcases were examined post-mortem. Histological lesions of pulmonary, hepatic and splenic erythrophagocytosis and haemosiderosis were found in 23 of 24 birds. Fifteen birds also had haemoglobin-like protein droplets within renal tubular epithelial cells. Despite consistent histological lesions, a cause of death could not be established. Virology, bacteriology and molecular tests for avian influenza, avian paramyxovirus-1, avipoxvirus,Chlamydia psittaci,Plasmodiumspp.,Babesiaspp.,Leucocytozoonspp. andToxoplasma gondiiwere negative. Tissue concentrations of a range of heavy metals (n = 4 birds) were consistent with low level exposure, while examination of proventricular contents and mucus failed to detect any marine biotoxins orClostridium botulinumtoxin. Hepatic concentrations of total polycyclic aromatic hydrocarbons (PAHs) (n = 5 birds) were similar to background concentrations of polycyclic aromatic hydrocarbons previously found in yellow-eyed penguins from the South Island of New Zealand, but there were significantly higher concentrations of 1-methylnapthelene and 2-methylnapthelene in the birds found dead in this mortality cluster. The biological significance of this finding is unclear. A temporal investigation of the epidemic did not indicate either a common source or propagative epidemic pattern. Although our investigation did not definitively implicate a toxic or infectious agent, we could not rule out causes such as toxic marine organisms or mycoplasmosis. Further investigations should therefore by carried out in the event of future mortality clusters. [ABSTRACT FROM AUTHOR]

C17-sphinganine analog mycotoxin (C17-SAMT) has been characterized as the contaminant responsible for the atypical toxicity reported in mussels from the Bizerte lagoon (northern Tunisia) over the past decade. C17-SAMT exhibited common symptoms of toxicity in mice, including flaccid paralysis and severe respiratory distress, followed by rapid death. To determine the potential health risks of this neurotoxin, we assessed its subchronic toxicity according to the recommendations of OCDE n° 407. The body weight and the structural changes of vital organs were recorded. Biochemical and hematological parameters were also quantified. Macroscopic observations showed that mice treated with 0.9, 9, and 90 µg/kg C17-SAMT had significantly reduced stomach weights, swollen and fragile intestines, and signs of nephritis with renal abscesses. Transaminase assays pointed out that exposure to C17-SAMT can lead to transaminitis. Above-average lactate dehydrogenase values were recorded in both the treated and satellite groups. Hematology data showed a significant reduction in red blood cell counts in high-dose-treated group. Reductions in hemoglobin and hematocrit were also recorded. Mean leukocyte counts were significantly elevated in the high-, mid-dose treated and satellite groups. At the microscopic level, we noted myocardial atrophy and hyperemia. In the lungs, we noted necrosis associated with macrophages perivascular infiltration and congestion. The kidneys showed mild inflammation and glomerular atrophy. The stomach exhibited mucosal atrophy, while a thin colon and distended small intestine were observed in high-dose-treated group. The liver was affected by vascular congestion, inflammatory infiltration, and lobular necrosis that evolved into acute hepatitis. Lesions, such as inflammatory infiltration and mild necrosis of the liver, cortical abscess with central necrosis in the kidney, and mild congestion of cardiac tissue were recorded in the satellite group. • Subchronic toxicity of the marine toxin C17_SAMT was assessed according to the recommendations of OCDE n° 407. • C17_SAMT significantly reduces stomach weight, swelling and friable bowel, and induces signs of nephritis and renal abscess. • Exposure to C17-SAMT can lead to transaminitis and a significant reduction in red blood. • We noted myocardial atrophy and congestion, pulmonary necrosis, mild inflammation, and renal glomerular atrophy. • We noted hepatic vascular congestion, inflammatory infiltrates, and lobular necrosis. [ABSTRACT FROM AUTHOR]

Marine toxins have a significant impact on seafood resources and human health. Up to date, mainly based on bioassays results, two genera of toxic microalgae, Gambierdiscus and Fukuyoa have been hypothesized to produce a suite of biologically active compounds, including maitotoxins (MTXs) and ciguatoxins (CTXs) with the latter causing ciguatera poisoning (CP) in humans. The global ubiquity of these microalgae and their ability to produce (un-)known bioactive compounds, necessitates strategies for screening, identifying, and reducing the number of target algal species and compounds selected for structural elucidation. To accomplish this task, a dereplication process is necessary to screen and profile algal extracts, identify target compounds, and support the discovery of novel bioactive chemotypes. Herein, a dereplication strategy was applied to a crude extract of a G. balechii culture to investigate for bioactive compounds with relevance to CP using liquid chromatography-high resolution mass spectrometry, in vitro cell-based bioassay, and a combination thereof via a bioassay-guided micro-fractionation. Three biologically active fractions exhibiting CTX-like and MTX-like toxicity were identified. A naturally incurred fish extract (Sphyraena barracuda) was used for confirmation where standards were unavailable. Using this approach, a putative I/C-CTX congener in G. balechii was identified for the first time, 44-methylgambierone was confirmed at 8.6 pg cell−1, and MTX-like compounds were purported. This investigative approach can be applied towards other harmful algal species of interest. The identification of a microalgal species herein, G. balechii (VGO920) which was found capable of producing a putative I/C-CTX in culture is an impactful advancement for global CP research. The large-scale culturing of G. balechii could be used as a source of I/C-CTX reference material not yet commercially available, thus, fulfilling an analytical gap that currently hampers the routine determination of CTXs in various environmental and human health-relevant matrices. [Display omitted] • Putative I/C-CTX-1 identified for the first time in a dinoflagellate extract. • Confirmation of 44-methylgambierone by LC-HRMS2 in G. balechii. • Interdisciplinary two-tiered approach for the dereplication of toxins. • Bio-guided fractionation of G. balechii extract by LC-HRMS and N2a-assay. [ABSTRACT FROM AUTHOR]

The dinoflagellate Prorocentrum belizeanum is responsible for the production of several toxins involved in the red tide phenomenon known as Diarrhetic Shellfish Poisoning (DSP). In this paper we report on the biosynthetic origin of an okadaic acid water-soluble ester derivative, DTX5c, on the basis of the spectroscopical analysis of 13C enriched samples obtained by addition of labelled sodium [l-13C], [2-13C] acetate to artificial cultures of this dinoflagellate.

It is well established that sea anemones comprise a rich source of cytolytic toxins. The present study reports the isolation and characterization of a cytolysin obtained from the sea anemone Heteractis magnifica collected in the Andaman Islands of the Indian Ocean. The crude extract was screened for hemolytic activity by a blood agar plate method and a 6-mm zone of clearance was observed after incubation. The hemolytic property of the crude extract, tested by the microtiter plate method, revealed positive results at concentrations as low as 120 ng/mL. Furthermore, it was favored by alkaline pH and was stable up to 60°C. On the other hand, the hemolytic effect was abolished by the addition of human serum. Purification steps involved ammonium sulfate precipitation and subsequent desalting by dialysis, followed by anion- and cation-exchange chromatographies. The purified fractions displayed the presence of a 19-kDa cytolysin when analyzed by SDS-PAGE. The conserved region of the cytolysin (with 303 bp) was amplified by RT-PCR and was sequenced. The sequence showed maximum homology (97%) with the already reported cytolysins from other sea anemone species.

Research in biosensing approaches as alternative techniques for food diagnostics for the detection of chemical contaminants and foodborne pathogens has increased over the last twenty years. The key component of such tests is the biorecognition element whereby polyclonal or monoclonal antibodies still dominate the market. Traditionally the screening of sera or cell culture media for the selection of polyclonal or monoclonal candidate antibodies respectively has been performed by enzyme immunoassays. For niche toxin compounds, enzyme immunoassays can be expensive and/or prohibitive methodologies for antibody production due to limitations in toxin supply for conjugate production. Automated, self-regenerating, chip-based biosensors proven in food diagnostics may be utilised as rapid screening tools for antibody candidate selection. This work describes the use of both single channel and multi-channel surface plasmon resonance (SPR) biosensors for the selection and characterisation of antibodies, and their evaluation in shellfish tissue as standard techniques for the detection of domoic acid, as a model toxin compound. The key advantages in the use of these biosensor techniques for screening hybridomas in monoclonal antibody production were the real time observation of molecular interaction and rapid turnaround time in analysis compared to enzyme immunoassays. The multichannel prototype instrument was superior with 96 analyses completed in 2 h compared to 12 h for the single channel and over 24 h for the ELISA immunoassay. Antibodies of high sensitivity, IC 50 's ranging from 4.8 to 6.9 ng/mL for monoclonal and 2.3–6.0 ng/mL for polyclonal, for the detection of domoic acid in a 1 min analysis time were selected. Although there is a progression for biosensor technology towards low cost, multiplexed portable diagnostics for the food industry, there remains a place for laboratory-based SPR instrumentation for antibody development for food diagnostics as shown herein. [ABSTRACT FROM AUTHOR]

Current regulation for marine toxins requires a monitoring method based on mass spectrometric analysis. This method is pre-targeted, hence after searching for pre-assigned masses, it identifies those compounds that were pre-defined with available calibrants. Therefore, the scope for detecting novel toxins which are not included in the monitoring protocol are very limited. In addition to this, there is a poor comprehension of the toxicity of some marine toxin groups. Also, the validity of the current approach is questioned by the lack of sufficient calibrants, and by the insufficient coverage by current legislation of the toxins reported to be present in shellfish. As an example, tetrodotoxin, palytoxin analogs, or cyclic imines are mentioned as indicators of gaps in the system that require a solid comprehension to assure consumers are protected. [ABSTRACT FROM AUTHOR]

A label-free electrochemical aptasensor was developed for selective detection of saxitoxin (STX). It is taking advantage of target-induced conformational change of an STX-specific aptamer when it binds to the toxin. A monolayer of octadecanethiol was deposited on a gold electrode, and then coated with a film of multiwalled carbon nanotubes (MWCNTs) to which the aptamer was covalently conjugated. Methylene blue (MB) was electrostatically anchored on carboxylated MWCNTs and used as the electrochemical indicator that produced a strong differential pulse voltammetric signal in the absence of target (STX). If, however, STX binds to its aptamer, this triggers a conformational change of the aptamer and results in the establishment of a barrier for heterogeneous electron transfer. The oxidation peak current of MB, acquired at −0.27 V (vs. Ag/AgCl), linearly decreases with increasing concentrations of STX in the 0.9 and 30 nM concentration range. The detection limit is 0.38 nM. Marine toxins that maybe present along with STX do not interfere even if they have a similar chemical structure. The assay was applied to the determination of STX in mussels samples and was found to be acceptably accurate. Hence, the method introduced here provides a rapid and sensitive tool for monitoring red tide pollution. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

Saxitoxin (STX) is a highly toxic marine neurotoxin produced by phytoplankton and a growing threat to ecosystems worldwide due to the spread of toxic algae. Although STX is an established sodium channel blocker, the overall profile of transcriptional levels in STX-exposed organisms has yet to be described. Here, we describe a toxicity assay and transcriptome analysis of the copepod Tigriopus japonicus exposed to STX. The half-maximal lethal concentration of STX was 12.35 μM, and a rapid mortality slope was evident at concentrations between 12 and 13 μM. STX induced changes in swimming behavior among the copepods after 10 min of exposure. In transcriptome analysis, gene ontology revealed that the genes involved in nervous system and gene expression were highly enriched. In addition, the congenital neurological disorder and nuclear factor erythroid 2-related factor 2–mediated oxidative stress pathways were identified to be the most significant in network analysis and toxicity pathway analysis, respectively. This study provides valuable information about the effects of STX and related transcriptional responses in T. japonicus. [Display omitted] • Saxitoxin exposure induced mortality and behavioral changes in Tigriopus japonicus. • Genes involved in neuron system-related pathways were deregulated. • NRF2-mediated oxidative stress response was predicted as the main toxicity pathway. [ABSTRACT FROM AUTHOR]

A new signal amplification strategy based on mesoporous carbon-enriched palladium nanostructure (MSC-PdNS) was designed for enzyme-free electrochemical immunoassay of brevetoxin B (BTB) in marine toxins. The assay was carried out on a BTB-bovine serum albumin-functionalized electrode by using monoclonal mouse anti-BTB-labeling MSC-PdNS as the signal-transduction tag. A competitive-type assay protocol was successfully introduced to develop a high-efficiency enzyme-free immunoassay accompanying the doped palladium nanostructure into MSC-PdNS toward reduction of H 2 O 2 . Under the optimal conditions, the catalytic current decreased with the increment of BTB concentration in the range from 0.01 to 10 ng mL −1 with a detection limit (LOD) of 5.0 pg mL −1 BTB at the 3 s blank criterion. The selectivity and precision were acceptable. In addition, the methodology was further validated for assaying spiked seafood samples, and consistent results between the electrochemical immunoassay and the referenced enzyme immunoassay were obtained. Importantly, the enzyme-free electrochemical immunoassay provides a promising approach for rapid screening of marine toxin because of its simplicity, low cost, sensitivity, specificity and without the need of sample pretreatment. [ABSTRACT FROM AUTHOR]

Current regulation for marine toxins requires a monitoring method based on mass spectrometric analysis. This method is pre-targeted, hence after searching for pre-assigned masses, it identifies those compounds that were pre-defined with available calibrants. Therefore, the scope for detecting novel toxins which are not included in the monitoring protocol are very limited. In addition to this, there is a poor comprehension of the toxicity of some marine toxin groups. Also, the validity of the current approach is questioned by the lack of sufficient calibrants, and by the insufficient coverage by current legislation of the toxins reported to be present in shellfish. As an example, tetrodotoxin, palytoxin analogs, or cyclic imines are mentioned as indicators of gaps in the system that require a solid comprehension to assure consumers are protected.

Neosaxitoxin (NeoSTX) is a specific reversible blocker of voltage gated sodium channels on excitable cells. In the last decade, it has been tested in a number of interesting clinical trials, however there is still little information available on mammalian toxicity. Rats were treated for 12 weeks with doses of 1, 3 or 6 μg/kg of subcutaneous NeoSTX. At weeks 12 and 17, animals were sacrificed and blood samples collected for hematological and biochemical analysis. Organs were harvested for weight determination and histopathological assessments. The lowest acute toxicity via the intraperitoneal (ip) route was (30.35 μg/kg) and there was no significant difference between intramuscular and subcutaneous routes (11.4 and 12.41 μg/kg). The NeoSTX adiministration did not produce lethality at week 12 and after five weeks of suspension. NeoSTX 6 μg/kg ip produced reductions (p < 0.05) in body weight and food intake, and increased blood level of total and direct bilirubin, GGT and SGOT at week 12; all of these were reversed in the recovery period. NeoSTX 1 and 3 μg/kg ip did not show significant changes with the control group. Histopathological presentations were normal in all groups. This study revealed that NeoSTX is safe in vivo, giving a reliable security margin for its use like a local anesthetic. [ABSTRACT FROM AUTHOR]

Marine organisms are an increasingly important source of novel metabolites, some of which have already inspired or become new drugs. In addition, many of these molecules show a high degree of novelty from a structural and/or pharmacological point of view. Structure determination is generally achieved by the use of a variety of spectroscopic methods, among which NMR (nuclear magnetic resonance) plays a major role and determination of the stereochemical relationships within every new molecule is generally the most challenging part in structural determination. In this communication, we have chosen okadaic acid as a model compound to perform a computational chemistry study to predict ¹H and 13C NMR chemical shifts. The effect of two different solvents and conformation on the ability of DFT (density functional theory) calculations to predict the correct stereoisomer has been studied. [ABSTRACT FROM AUTHOR]

Saxitoxin (STX), is one of the most dangerous and widespread paralytic shellfish toxins, causing a severe threat to the ecosystem and human health. So, it is important and highly essential to develop novel techniques for STX detection in a convenient, desirable, and low-cost manner. Herein, this study developed an electrolyte-insulator-semiconductor (EIS) sensor covered with a layer-by-layer prepared, positively-charged weak polyelectrolyte layer of poly (allylamine hydrochloride) (PAH) for the label-free detection of STX. The specific aptamer (Apt) sensitive to STX was electrostatically adsorbed onto the PAH layer. This leads to a preferentially flat orientation of the Apt within the Debye length, thus yielding a reduced charge-screening influence and a higher sensor signal. Each step of sensor surface modification, i.e. PAH adsorption, immobilization of Apt, and attachment of STX, was monitored by capacitance-voltage (C - V) and constant-capacitance (ConCap) measurements. Furthermore, atomic force microscopy (AFM) was employed to characterize the surface morphology and roughness of the PAH layer. Fluorescence microscopy was used to confirm the effective immobilization of Apt onto the PAH-modified EIS sensor. The results showed that the detection range of this aptasensor for STX detection was 0.5–100 nM and the detection limit was as low as 0.05 nM. Furthermore, this aptasensor showed good selectivity and 9 days' stability. The mussel tissue extraction test suggested that this aptasensor can be used to detect STX in real samples. This aptasensor provides a convenient approach for moderate, rapid, and label-free detection of marine biological toxins. [Display omitted] • An electrolyte-insulator-semiconductor (EIS) sensor was applied for the label-free detection of saxitoxin. • The detection range for saxitoxin was 0.1–100 nM and a low detection limit of 0.01 nM. • The sensor was applied for real food sample measurement and showed good selectivity and 14 day's stability. [ABSTRACT FROM AUTHOR]

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]

Tetrodotoxin (TTX), paralytic shellfish poisons (PSP) and ciguateric toxins (CTXs) are the most important marine toxins causing seafood poisoning for humans. In this report, the poisoning cases and the causative species of marine toxins in Taiwan in recent year were reviewed. TTX poisoning cases in Taiwan were usually caused by puffer processed products such as dried dressed puffer fillet and puffer roe, followed by toxic puffer, gastropod and goby. PSP poisoning cases in Taiwan were usually caused by gastropod and shellfish contaminated from dinoflagellate Alexandrium minutum, and the main toxins were identified as saxitoxin, neo saxitoxin and gonyautoxin . For CTXs poisoning cases, the causative species in Taiwan included snapper, moray eel and sardine, and the suspected toxins were implicated as ciguatoxin and palytoxin. Accordingly, the system of monitoring and risk warning for marine toxins in Taiwan was formulated. The sampling and management measures are suggested for fishery products suspected with each toxin. The monitoring objectives are toxins and species inspections, and the analytic techniques including mouse bioassay, HPLC, GC-MS and LC-MS/MS can be employed for toxins investigation. Furthermore, the risk warning system should focus on toxic algae, shellfish and gastropod, which implicate PSP toxins. An alarm can be declared when the number of dinoflagellates is higher than 500 cells/L in the fish farm. The monitoring standards, control program and warning model of marine toxins should be established as soon as possible. [ABSTRACT FROM AUTHOR]

The marine toxin, okadaic acid (OA) is produced by dinoflagellates of the genera Prorocentrum and Dinophysis and is the causative agent of the syndrome known as diarrheic shellfish poisoning. In addition, OA acts as both a tumor promoter, attributed to OA-induced inhibition of protein phosphatases as well as an inducer of apoptosis. To better understand the potentially divergent toxicological profile of OA, the concentration-dependent cytotoxicity and alterations in gene expression on the human liver tumor cell line HepG2 upon OA exposure were determined using RNA microarrays, DNA fragmentation, and cell proliferation assays as well as determinations of cell detachment and cell death in different concentrations of OA. mRNA expression was quantified for approximately 15,000 genes. Cell attachment and proliferation were both negatively correlated with OA concentration. Detached cells displayed necrotic DNA signatures but apoptosis also was broadly observed. Data suggest that OA has a concentration dependent effect on cell cycle, which might explain the divergent effects that at low concentration OA stimulates genes involved in the cell cycle and at high concentrations it stimulates apoptosis. [ABSTRACT FROM AUTHOR]

The dinoflagellate Prorocentrum belizeanum is responsible for the production of several toxins involved in the red tide phenomenon known as Diarrhetic Shellfish Poisoning (DSP). In this paper we report on the biosynthetic origin of an okadaic acid water-soluble ester derivative, DTX5c, on the basis of the spectroscopical analysis of 13C enriched samples obtained by addition of labelled sodium [1-13C], [2-13C] acetate to artificial cultures of this dinoflagellate. [ABSTRACT FROM AUTHOR]

Abstract: In this work, a highly selective sample cleanup procedure that combining molecular imprinting technique (MIT) and solid phase extraction (SPE) was developed for the isolation of domoic acid (a fascinating marine toxin) from seafood samples. The molecular imprinting polymer (MIP) for domoic acid was prepared using 1,3,5-pentanetricarboxylic acid as the template molecule instead of domoic acid. 4-Vinyl pyridine was used as the functional monomer and ethylene glycol dimethacrylate was used as the cross-linking monomer. The obtained imprinted polymer showed high affinity to domoic acid and was used as selective sorbent for the SPE of domoic acid from seafood samples. An off-line molecularly imprinted solid phase extraction (MISPE) method followed by high-performance liquid chromatography (HPLC) with diode-array detection for the detection of domoic acid was also established. Good linearity was obtained from 0.5mgL−1 to 25mgL−1 (R 2 >0.99) with a quantitation limit of 0.1mgL−1, which was sufficient to determine domoic acid at the maximum level permitted by several authorities. The mean recoveries of domoic acid from mussel extracts were 93.4–96.7%. It was demonstrated that the proposed MISPE–HPLC method could be applied to direct determination of domoic acid from seafood samples. [Copyright &y& Elsevier]

Abstract: This review considers the ethical and technical problems currently associated with employing mouse bioassays for marine-toxin analysis and the challenges and the difficulties that alternative methods must overcome before being deemed applicable for implementation into a regulatory monitoring regime. We discuss proposed alternative methods, classified as functional, immunological and analytical, for well-established European toxins as well as emerging toxins in European waters, highlighting their advantages and disadvantages. We also consider emerging tools and technologies for future toxin analysis. Even though regulatory bodies have recently recommended analytical methods for a number of toxins, there is still scope for functional and immunological methods in rapid screening and detecting emerging toxins. Future developments foreseen in the analysis of marine biotoxins are multiplex-based analysis, miniaturization and portability for on-site testing. However, the longstanding lack of reference materials and standards continues to pose a severe limitation on progress in development, validation and therefore implementation of any alternative method based on the criteria stipulated by European Union legislation. [Copyright &y& Elsevier]