Ciguatoxin Detection Methods and High‐Throughput Assays

Inserra, Marco; Lavrukhina, Yelena; Jones, Alun; Lewis, Richard J.; Vetter, Irina

doi:10.1002/9781118992685.ch15

Cited by 3 publications

(11 citation statements)

References 96 publications

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“…CTXs bind quasi-irreversibly to voltage-sensitive sodium channels, enhancing sodium influx into cells, causing them to open at the normal cell-resting membrane potential, thereby impeding normal function. In addition, CTXs also inhibit neuronal potassium channels, which is likely to act in concert with effects on voltage-gated sodium channels to increase neuronal excitability [ 2 , 3 ]. Evaluation of cell viability forms the basis of CTX determination in CBAs [ 1 ].…”

Section: Detection and Quantification Of Ciguatoxinsmentioning

confidence: 99%

“…The CBA-N2a is widely used today, and its development by Manger et al [ 74 ] is one of the most important milestones of replacing MBA in modern laboratories. The assay is based on the colorimetric detection of metabolically active N2a cells treated with CTX extract in the presence of ouabain/veratridine [ 74 , 75 , 76 ]; it expresses negatively the concentration of various voltage-gated sodium channel toxins and assesses cell death [ 3 ]. CTXs have no cytotoxic effect on N2a cells, therefore their detection requires addition of veratridine (a sodium-channel-activator that have a different binding site than CTXs) and ouabain (a sodium/potassium ion ATPase inhibitor).…”

Section: Detection and Quantification Of Ciguatoxinsmentioning

confidence: 99%

“…Ciguatera fish poisoning (CFP), currently the most common marine biotoxin food poisoning worldwide, is a non-bacterial foodborne disease associated with consumption of circumtropical fish and marine invertebrates that are contaminated with polyether sodium channel activator neurotoxins (ciguatoxins, CTXs) [ 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 ]. CTXs are a family of heat-stable and lipid-soluble compounds that cannot be degraded by normal cooking.…”

Section: Introductionmentioning

confidence: 99%

“…CFP is known in tropical regions for centuries, and it is an increasing risk of food poisoning worldwide; it occurs now in non-endemic areas due to international trade of fish and fish products and the expansion of the geographic ranges of dinoflagellates as a likely result of global warming [ 12 , 16 , 17 ]. Intoxication by CTXs may cause neurological, gastrointestinal, and cardiovascular symptoms depending on the amount and type of the toxin ingested [ 1 , 2 , 3 , 4 , 5 , 6 , 7 , 18 ], and occasionally in severe cases, CFP can be fatal [ 19 , 20 ]. P-CTX-1 possesses risk to human health at concentrations higher than 0.022–0.1 ng g −1 in fish flesh [ 8 , 21 ].…”

Section: Introductionmentioning

confidence: 99%

“…In extreme cases, such as for the liver of a large moray eel caught in Kiribati, toxicity can be as high as 539 ng g ‒1 , 50,000 times higher than the accepted safety level of 0.01 ng g ‒1 [ 23 ]. The symptoms of CFP were first described by Captain James Cook and Don Antonio Parra in the 17th century during their exploration of the Pacific Ocean and Caribbean Sea, respectively [ 3 , 24 ]. CFP was finally linked to dinoflagellates in 1977 [ 25 ].…”

Section: Introductionmentioning

confidence: 99%

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Advances in Detecting Ciguatoxins in Fish

Pasinszki

Lako

Dennis

2020

Toxins

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Ciguatera fish poisoning (CFP) is currently the most common marine biotoxin food poisoning worldwide, associated with human consumption of circumtropical fish and marine invertebrates that are contaminated with ciguatoxins. Ciguatoxins are very potent sodium-channel activator neurotoxins, that pose risks to human health at very low concentrations (>0.01 ng per g of fish flesh in the case of the most potent Pacific ciguatoxin). Symptoms of CFP are nonspecific and intoxication in humans is often misdiagnosed. Presently, there is no medically approved treatment of ciguatera. Therefore, to mitigate the risks of CFP, reliable detection of ciguatoxins prior to consumption of fish tissue is acutely needed, which requires application of highly sensitive and quantitative analytical tests. During the last century a number of methods have been developed to identify and quantify the concentration of ciguatoxins, including in vivo animal assays, cell-based assays, receptor binding assays, antibody-based immunoassays, electrochemical methods, and analytical techniques based on coupling of liquid chromatography with mass spectrometry. Development of these methods, their various advantages and limitations, as well as future challenges are discussed in this review.

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Section: Detection and Quantification Of Ciguatoxinsmentioning

confidence: 99%

Section: Detection and Quantification Of Ciguatoxinsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Advances in Detecting Ciguatoxins in Fish

Pasinszki

Lako

Dennis

2020

Toxins

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Faster ciguatoxin extraction methods for toxicity screening

Loeffler,

Spielmeyer

2024

Sci Rep

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Ciguatera poisoning (CP) is a severe global public health problem caused by the consumption of seafood products contaminated with ciguatoxins (CTXs). The growing demand for seafood products requires high-throughput testing for CTX-susceptible seafood, however complex extraction and slow cleanup methods inhibit this goal. Herein, several methods for extracting CTXs from fish tissue were established and compared; these methods are sensitive, specific, and valid while achieving higher sample extraction throughput than currently established protocols. The trial fish material was generated from multiple species, with different physical conditions (wet and freeze-dried tissue), and naturally contaminated with various CTXs (i.e., CTX-1B, CTX-3C, and C-CTX-1), thus ensuring these methods are robust and broadly applicable. The extraction methods used were based on mechanical maceration with acetone or methanol or enzymatic digestion followed by acetone and ethyl acetate extraction. Crude extracts were investigated for CTX-like toxicity using an in vitro mouse neuroblastoma (N2a) cell-based assay (CBA). Among the three methods, there was no significant difference in toxin estimates (p = 0.219, two-way ANOVA), indicating their interchangeability. For speed (> 16 samples/day), accuracy (100%), and CTX analog retention confirmation by liquid chromatography-tandem mass spectrometry (LC‒MS/MS), the preferred extraction methods were both methanol and enzyme-based. All extraction methods post hoc confirmation of CTX analogs successfully met international seafood market-based CTX contaminant guidance. These methods can drastically increase global CTX screening capabilities and subsequently relieve sample processing bottlenecks, inhibiting environmental and human health-based CTX analysis.

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Ciguatera Fish Poisoning in the Caribbean Sea and Atlantic Ocean: Reconciling the Multiplicity of Ciguatoxins and Analytical Chemistry Approach for Public Health Safety

Pottier

Lewis

Vernoux

2023

Toxins

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Ciguatera is a major circumtropical poisoning caused by the consumption of marine fish and invertebrates contaminated with ciguatoxins (CTXs): neurotoxins produced by endemic and benthic dinoflagellates which are biotransformed in the fish food-web. We provide a history of ciguatera research conducted over the past 70 years on ciguatoxins from the Pacific Ocean (P-CTXs) and Caribbean Sea (C-CTXs) and describe their main chemical, biochemical, and toxicological properties. Currently, there is no official method for the extraction and quantification of ciguatoxins, regardless their origin, mainly due to limited CTX-certified reference materials. In this review, the extraction and purification procedures of C-CTXs are investigated, considering specific objectives such as isolating reference materials, analysing fish toxin profiles, or ensuring food safety control. Certain in vitro assays may provide sufficient sensitivity to detect C-CTXs at sub-ppb levels in fish, but they do not allow for individual identification of CTXs. Recent advances in analysis using liquid chromatography coupled with low- or high-resolution mass spectrometry provide new opportunities to identify known C-CTXs, to gain structural insights into new analogues, and to quantify C-CTXs. Together, these methods reveal that ciguatera arises from a multiplicity of CTXs, although one major form (C-CTX-1) seems to dominate. However, questions arise regarding the abundance and instability of certain C-CTXs, which are further complicated by the wide array of CTX-producing dinoflagellates and fish vectors. Further research is needed to assess the toxic potential of the new C-CTX and their role in ciguatera fish poisoning. With the identification of C-CTXs in the coastal USA and Eastern Atlantic Ocean, the investigation of ciguatera fish poisoning is now a truly global effort.

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Ciguatoxin Detection Methods and High‐Throughput Assays

Cited by 3 publications

References 96 publications

Advances in Detecting Ciguatoxins in Fish

Advances in Detecting Ciguatoxins in Fish

Faster ciguatoxin extraction methods for toxicity screening

Ciguatera Fish Poisoning in the Caribbean Sea and Atlantic Ocean: Reconciling the Multiplicity of Ciguatoxins and Analytical Chemistry Approach for Public Health Safety

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