Publications by authors named "Rambla-Alegre M"

Freshwater snails of the genus Pomacea, have been introduced worldwide with well-documented ecological and economic impacts. In rice, a crucial crop for global food security, apple snails have become a major pest. Chemical molluscicides, the most effective method of control, raise concerns due to their ecotoxicity.

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  • Pufferfish, known for being highly toxic due to neurotoxins like tetrodotoxins and paralytic shellfish toxins, poses a serious risk to human health and has caused many poisoning incidents.
  • A study analyzed tissue extracts from two pufferfish species collected from the Spanish Mediterranean, revealing no toxicity in Sphoeroides pachygaster but significant toxicity in the liver of Lagocephalus lagocephalus specimens.
  • High-performance liquid chromatography analysis confirmed the presence of paralytic shellfish toxins, particularly saxitoxin and decarbamoylsaxitoxin, in L. lagocephalus, highlighting a food safety concern due to the potential risks associated with consuming this toxic fish in the region. *
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  • Tetrodotoxin (TTX) is a dangerous marine neurotoxin linked to various poisoning cases and fatalities, with over 30 known analogues, but their toxic effects are not fully understood.
  • This study focused on determining the toxicity equivalency factors (TEFs) of five TTX analogues by examining how they affect voltage-gated sodium channels in Neuro-2a cells using an automated patch clamp method.
  • Results showed that all TTX analogues were less toxic than TTX, and the derived TEFs can effectively convert LC-MS/MS data from pufferfish samples into relevant toxicological insights, emphasizing the potential of this method for food safety and health protection.
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Spirolides (SPXs), gymnodimines (GYMs), and pinnatoxins (PnTXs) have been detected in shellfish from the northwestern Mediterranean coast of Spain. Several samples of bivalves were collected from Fangar Bay and Alfacs Bay in Catalonia over a period of over 7 years (from 2015 to 2021). Shellfish samples were analyzed for cyclic imines (CIs) on an LC1200 Agilent and 3200 QTrap triple-quadrupole mass spectrometer.

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Tetrodotoxin (TTX) is a potent marine neurotoxin involved in poisoning cases, especially through the consumption of puffer fish. Knowledge of the toxicity equivalency factors (TEFs) of TTX analogues is crucial in monitoring programs to estimate the toxicity of samples analyzed with instrumental analysis methods. In this work, TTX analogues were isolated from the liver of a individual caught on South Crete coasts.

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Ciguatoxins (CTXs) are marine toxins produced by microalgae of the genera Gambierdiscus and Fukuyoa, which are transferred through the food webs, reaching humans and causing a poisoning known as ciguatera. The cell-based assay (CBA) is commonly used for their detection because of its high sensitivity and the provided toxicological information. However, matrix effects may interfere in the CBA.

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Ciguatera Poisoning (CP) is caused by consumption of fish or invertebrates contaminated with ciguatoxins (CTXs). Presently CP is a public concern in some temperate regions, such as Macaronesia (North-Eastern Atlantic Ocean). Toxicity analysis was performed to characterize the fish species that can accumulate CTXs and improve understanding of the ciguatera risk in this area.

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Intense agricultural activities are performed in the Ebro River Delta (NE Spain) with extensive use of pesticides. Medium to highly polar pesticides have not been studied intensively in sediments despite its larger use in the recent years. This work aimed at assessing the occurrence of 69 pesticides, including medium to highly polar compounds, in sediments collected from drainage and irrigation channels of the Ebro River Delta during the main rice growing season.

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Cyclodextrins, cyclic oligomers that form a conical structure with an internal cavity, are proposed as new and sustainable materials for passive sampling of lipophilic marine toxins. Two applicability scenarios have been tested. First, disks containing β-cyclodextrin-hexamethylene diisocyanate (β-CD-HDI) and β-cyclodextrin-epichlorohydrin (β-CD-EPI) polymers were immersed in Prorocentrum lima cultures for different days (2, 12 and 40).

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Delta ecosystems are areas of high ecologic and economic values, where wildlife commonly shares the territory with intensive agricultural activities, particularly, rice cultivation and seafood production. This work aimed at evaluating the occurrence of a wide spectrum of pesticides and transformation products in the water of irrigation and drainage channels of the Ebro River Delta (NE Spain) during the main rice-growing season, when pesticide application is at its peak. Furthermore, the impact that these contaminants may have on local ecosystems and seafood production activities was assessed.

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  • Recent research has advanced understanding of dinoflagellate taxonomy and distribution, particularly focusing on their bioactive compounds related to ciguatera poisoning (CP).
  • Ciguatoxins and maitotoxins, notably maitotoxin-4, are highly toxic metabolites linked to CP and have been increasingly identified in various regions, including the Canary Islands and Madeira archipelago.
  • This study emphasizes the importance of characterizing toxin profiles across different species and geographic areas, revealing that these profiles may vary significantly by species and location.
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Paralytic shellfish poisoning (PSP) episodes cause important economic impacts due to closure of shellfish production areas in order to protect human health. These closures, if are frequent and persistent, can seriously affect shellfish producers and the seafood industry, among others. In this study, we have developed an alternative processing method for bivalves with PSP content above the legal limit, which allows reducing toxicity to acceptable levels.

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Tetrodotoxin (TTX) is a potent neurotoxin responsible for many food poisoning incidents and some fatalities. Although mainly associated with the consumption of pufferfish, in recent years, TTX has been found in shellfish, particularly in Europe. In this work, a magnetic bead (MB)-based colorimetric immunoassay was applied to the detection of TTX in Pacific oysters (Crassostrea gigas), razor clams (Solen marginatus) and mussels (Mytilus galloprovincialis).

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The importance of ciguatoxins (CTXs) in seafood safety and their emerging occurrence in locations far away from tropical areas highlight the need for simple and low-cost methods for the sensitive and rapid detection of these potent marine toxins to protect seafood consumers. Herein, an electrochemical immunosensor for the detection of CTXs is presented. A sandwich configuration is proposed, using magnetic beads (MBs) as immobilization supports for two capture antibodies, with their combination facilitating the detection of CTX1B, CTX3C, 54-deoxyCTX1B, and 51-hydroxyCTX3C.

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One of the major challenges currently faced is to develop systematic ways of addressing chemical mixtures in environmental assessment. With this purpose, a simple, rapid, and sensitive method for the detection and quantification of a mixture of relevant contaminants in molluscs has been developed. The method is based on QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) and Ultra-High Performance Liquid Chromatography-High Resolution Mass Spectrometry (UHPLC-HRMS).

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Article Synopsis
  • Tetrodotoxin (TTX) is a harmful neurotoxin found in some fish and shellfish in European waters, raising concerns about consumer health.
  • A study collected samples of bivalves and gastropods from the Spanish coast and analyzed them for TTX using advanced testing methods, finding no TTX levels above the safety threshold of 10 µg/kg.
  • Although the study didn't find any TTX in the samples, further research with more frequent sampling is suggested to better assess the risks associated with TTX in these seafood products.
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This study aimed to assess the bioaccessibility of different marine biotoxins in naturally contaminated shellfish and fish gonads using an in vitro digestion methodology. In general, hydrophilic toxins (domoic acid, paralytic shellfish poisoning toxins and tetrodotoxins) showed higher bioaccessibility than lipophilic ones (okadaic acid and azaspiracids). The bioaccessibility of toxins from the okadaic acid group ranged from 69% (raw European razor clams) to 74% (raw donax clams).

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  • - Two juvenile Lagocephalus sceleratus fish were caught in the North Aegean Sea in 2017, and a new detection tool for tetrodotoxins (TTXs), which are harmful neurotoxins, was tested on them.
  • - The detection tool found TTXs in both fish, with some tissues containing toxin levels near or above 2 mg/kg, challenging the earlier assumption that these juveniles were non-toxic.
  • - This is the first report of high TTX levels in small Lagocephalus sceleratus, raising concerns about consumer safety as these toxic fish can be confused with edible species, emphasizing the need for accessible TTX detection tools to ensure seafood safety.
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Bivalves have proved to be useful bioindicators for environmental pollution. In the present study, mussels (Mytilus galloprovincialis), cockles (Cerastoderma edule), and razor shells (Solen marginatus) were collected in the Ebro Delta, an extensive area devoted to rice farming and affected by pesticide pollution, from April to July, the heaviest rice field treatment period. Possible effects of pollution were assessed through biochemical markers (carboxylesterase (CE), antioxidant and neurotoxicity-related enzymes, and lipid peroxidation levels).

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In August 2014, a puffer fish poisoning incidence resulting in one fatality was reported in New Caledonia. Although tetrodotoxin (TTX) intoxication was established from the patients' signs and symptoms, the determination of TTX in the patient's urine, serum or plasma is essential to confirm the clinical diagnosis. To provide a simple cost-effective rapid screening tool for clinical analysis, a maleimide-based enzyme-linked immunosorbent assay (mELISA) adapted for the determination of TTX contents in human body fluids was assessed.

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Cyclic imines constitute a quite recently discovered group of marine biotoxins that act on neural receptors and that bioaccumulate in seafood. They are grouped together due to the imino group functioning as their common pharmacore, responsible for acute neurotoxicity in mice. Cyclic imines (CIs) have not been linked yet to human poisoning and are not regulated in the European Union (EU), although the European Food Safety Authority (EFSA) requires more data to perform conclusive risk assessment for consumers.

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  • Rapid immunoassay methods have been developed to detect diarrhetic shellfish poisoning (DSP) toxins in seawater, essential for protecting public health and the shellfish industry.* ! -
  • These assays specifically target okadaic acid (OA) and two related toxins, achieving a detection limit of about 1 ng OA equiv./mL in seawater, and have been tested on samples from Catalan harbors and the Galician Rias.* ! -
  • Findings show that OA levels in seawater correlate with Dinophysis cell counts, but with a delay of 1-2 weeks, indicating that these immunoassays could serve as effective, high-throughput monitoring tools for DSP toxins.* !
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The recent detection of tetrodotoxins (TTXs) in puffer fish and shellfish in Europe highlights the necessity to monitor the levels of TTXs in seafood by rapid, specific, sensitive and reliable methods in order to protect human consumers. A previous immunoassay for TTX detection in puffer fish, based on the use of self-assembled monolayers (SAMs) for the immobilization of TTX on maleimide plates (mELISA), has been modified and adapted to the analysis of oyster and mussel samples. Changing dithiol for cysteamine-based SAMs enabled reductions in the assay time and cost, while maintaining the sensitivity of the assay.

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The recent detection of tetrodotoxins (TTXs) in European fish and shellfish has emphasized the urgent need to develop specific, selective, rapid and easy-to-use methods for their detection to assess the potential risk posed to human health. For this purpose, a dithiol self-assembled monolayer (SAM)-based immunoassay previously performed on maleimide plates (mELISA) has been adapted to gold electrode arrays for the development of an electrochemical immunosensor for TTX. The electrochemical SAM-based immunosensor designed herein, provided an oriented, stable and spaced sensing platform for the determination of TTX, attaining a limit of detection of 2.

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Severe food poisoning events after the consumption of sharks have been reported since the 1940s; however, there has been no clear understanding of their cause. Herein, we report for the first time the presence of ciguatoxins (CTXs) in sharks. The identification by mass spectrometry of CTXs, including two new analogues, in a bull shark (Carcharhinus leucas) that was consumed by humans, causing the poisoning and death of 11 people in Madagascar in 2013 is described.

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