Alternative Methods for the Detection of Emerging Marine Toxins: Biosensors, Biochemical Assays and Cell-Based Assays

Reverté, Laia; Soliño, Lucía; Carnicer, Olga; Diogène, Jorge; Campàs, Mònica

doi:10.3390/md12125719

Cited by 79 publications

(39 citation statements)

References 226 publications

(295 reference statements)

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“…The neuroblastoma cell-based assay (CBA-N2a) is widely used for the detection of a wide range of marine neurotoxins active on the VGSCs (Caillaud et al, 2010;Nicolas et al, 2014;Reverté et al, 2014). This toxicological assay classically uses a combination of ouabain (blocker of the Na + /K + ATPase pump) and veratridine (inductor of permanent activation of VGSCs), at selected concentrations, for the specific detection of CTXs (Manger et al, 1993;Cañ ete and Diogè ne, 2008;Caillaud et al, 2009).…”

Section: Neuroblastoma Cell-based Assays (Cba-n2a)mentioning

confidence: 99%

Evidence of the bioaccumulation of ciguatoxins in giant clams (Tridacna maxima) exposed to Gambierdiscus spp. cells

et al. 2016

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Ciguatera Fish Poisoning (CFP) is a foodborne disease classically related to the consumption of tropical coral reef fishes contaminated with ciguatoxins (CTXs), neurotoxins produced by dinoflagellates of the Gambierdiscus genus. Severe atypical ciguatera-like incidents involving giant clams, a marine resource highly consumed in the South Pacific, are also frequently reported in many Pacific Islands Countries and Territories. The present study was designed to assess the ability of giant clams to accumulate CTXs in their tissues and highlight the potential health risks associated with their consumption. Since giant clams are likely to be exposed to both free-swimming Gambierdiscus cells and dissolved CTXs in natural environment, ex situ contamination experiments were conducted as follows: giant clams were exposed to live or lyzed cells of TB92, a highly toxic strain of G. polynesiensis containing 5.83±0.85pg P-CTX-3C equiv.cellvs. HIT0, a weakly toxic strain of G. toxicus containing only (2.05±1.16)×10pg P-CTX-3C equiv.cell, administered over a 48h period at a concentration of 150cellsmL. The presence of CTXs in giant clams tissues was further assessed using the mouse neuroblastoma cell-based assay (CBA-N2a). Results showed that giant clams exposed to either lyzed or live cells of TB92 were able to bioaccumulate CTXs at concentrations well above the safety limit recommended for human consumption, i.e. 3.28±1.37 and 2.92±1.03ng P-CTX-3C equiv.g flesh (wet weight), respectively, which represented approximately 3% of the total toxin load administered to the animals. In contrast, giant clams exposed to live or lyzed cells of HIT0 were found to be free of toxins, suggesting that in the nature, the risk of contamination of these bivalves is established only in the presence of highly toxic blooms of Gambierdiscus. Liquid chromatography-mass spectrometry (LC-MS/MS) analyses confirmed CBA-N2a results and also revealed that P-CTX-3B was the major CTX congener retained in the tissues of giant clams fed with TB92 cells. To the best of our knowledge, this study is the first to provide evidence of the bioaccumulation of Gambierdiscus CTXs in giant clams and confirms that these bivalve molluscs can actually constitute another pathway in ciguatera poisonings. While most monitoring programs currently focus on fish toxicity, these findings stress the importance of a concomitant surveillance of these marine invertebrates in applicable locations for an accurate assessment of ciguatera risk.

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Section: Neuroblastoma Cell-based Assays (Cba-n2a)mentioning

confidence: 99%

Evidence of the bioaccumulation of ciguatoxins in giant clams (Tridacna maxima) exposed to Gambierdiscus spp. cells

et al. 2016

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“…Neuroblastoma cell-based assay accomplishes very well the last two requirements. Neuroblastoma cell-based assay can detect the effect of CTXs and MTXs analogues at low concentrations [49][50][51][52] and also other possible novel toxins with the same effect on sodium channels [53]. The drawback of this is that it cannot attribute the CTX-like or MXT-like effect to one specific molecule [52].…”

Section: Discussionmentioning

confidence: 99%

Toxicity Characterisation of Gambierdiscus Species from the Canary Islands

Rossignoli

Tudó

Bravo

et al. 2020

Toxins

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In the last decade, several outbreaks of ciguatera fish poisoning (CFP) have been reported in the Canary Islands (central northeast Atlantic Ocean), confirming ciguatera as an emerging alimentary risk in this region. Five Gambierdiscus species, G. australes, G. excentricus, G. silvae, G. carolinianus and G. caribaeus, have been detected in macrophytes from this area and are known to produce the ciguatoxins (CTXs) that cause CFP. A characterization of the toxicity of these species is the first step in identifying locations in the Canary Islands at risk of CFP. Therefore, in this study the toxicity of 63 strains of these five Gambierdiscus species were analysed using the erythrocyte lysis assay to evaluate their maitotoxin (MTX) content. In addition, 20 of the strains were also analysed in a neuroblastoma Neuro-2a (N2a) cytotoxicity assay to determine their CTX-like toxicity. The results allowed the different species to be grouped according to their ratios of CTX-like and MTX-like toxicity. MTX-like toxicity was especially high in G. excentricus and G. australes but much lower in the other species and lowest in G. silvae. CTX-like toxicity was highest in G. excentricus, which produced the toxin in amounts ranging between 128.2 ± 25.68 and 510.6 ± 134.2 fg CTX1B equivalents (eq) cell−1 (mean ± SD). In the other species, CTX concentrations were as follows: G. carolinianus (100.84 ± 18.05 fg CTX1B eq cell−1), G. australes (31.1 ± 0.56 to 107.16 ± 21.88 fg CTX1B eq cell−1), G. silvae (12.19 ± 0.62 to 76.79 ± 4.97 fg CTX1B eq cell−1) and G. caribaeus (<LOD to 90.37 ± 15.89 fg CTX1B eq cell−1). Unlike the similar CTX-like toxicity of G. australes and G. silvae strains from different locations, G. excentricus and G. caribaeus differed considerably according to the origin of the strain. These differences emphasise the importance of species identification to assess the regional risk of CFP.

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“…Chromatographic techniques coupled with several detection methods are powerful and accurate analysis tools, and are routinely used as reference methods for many marine toxins. However, the required instrumentation is expensive and requires trained personnel, and to address these shortcomings, the European Commission encourages the development and use of alternative or complementary methods [5], which are usually based on a functional or structural recognition of the toxin [6]. Cell-based assays (CBAs) are easy to perform, give an overall view of the toxicity of a sample and can detect the presence of unknown toxins.…”

Section: Marine Toxinsmentioning

confidence: 99%

Magnetic Beads in Marine Toxin Detection: A Review

2019

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Due to the expanding occurrence of marine toxins, and their potential impact on human health, there is an increased need for tools for their rapid and efficient detection. We give an overview of the use of magnetic beads (MBs) for the detection of marine toxins in shellfish and fish samples, with an emphasis on their incorporation into electrochemical biosensors. The use of MBs as supports for the immobilization of toxins or antibodies, as signal amplifiers as well as for target pre-concentration, is reviewed. In addition, the exploitation of MBs in Systematic Evolution of Ligands by Exponential enrichment (SELEX) for the selection of aptamers is presented. These MB-based strategies have led to the development of sensitive, simple, reliable and robust analytical systems for the detection of toxins in natural samples, with applicability in seafood safety and human health protection.

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Alternative Methods for the Detection of Emerging Marine Toxins: Biosensors, Biochemical Assays and Cell-Based Assays

Cited by 79 publications

References 226 publications

Evidence of the bioaccumulation of ciguatoxins in giant clams (Tridacna maxima) exposed to Gambierdiscus spp. cells

Evidence of the bioaccumulation of ciguatoxins in giant clams (Tridacna maxima) exposed to Gambierdiscus spp. cells

Toxicity Characterisation of Gambierdiscus Species from the Canary Islands

Magnetic Beads in Marine Toxin Detection: A Review

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