A cytotoxicity assay for the detection and differentiation of two families of shellfish toxins

Flanagan, Andrew F.; Callanan, K.R.; Donlon, John; Palmer, R.; Forde, A. M.; Kane, Marian

doi:10.1016/s0041-0101(00)00241-5

Cited by 51 publications

(23 citation statements)

References 18 publications

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“…T. radiatus and T. rudolphi showed medium toxicity against the normal cell lines at 1.87 and 1.54 mg/mL, respectively, whereas the control didn't show any considerable cell death. The morphological changes observed in the cells are concurrent with the previous reports on the cell morphology implementation of OA type toxins and azaspiracid in shellfish [37]. Likewise, Arokiyaraj et al [38] isolated the triterpenes from the leaves of Justicia gendarussa and established the maximum inhibition (85%) against human lymphocyte proliferation.…”

Section: Discussionsupporting

confidence: 81%

Investigation on neurotoxin of sea snail meat

Mohan¹,

Giji²,

Pachaiyappan³

et al. 2016

J Coast Life Med

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Section: Discussionsupporting

confidence: 81%

Investigation on neurotoxin of sea snail meat

Mohan¹,

Giji²,

Pachaiyappan³

et al. 2016

J Coast Life Med

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“…It has been postulated that some AZP toxins can inhibit protein phosphatases (PP), yet other scientists question this (Flanagan et al, 2001;twiner et al, 2005. In vitro experiments with mammalian cell lines showed alterations in the cytoskeletal structure (Roman et al, 2002;twiner et al, 2005;Ronzitti et al, 2007;Vilarino et al, 2006;Paredes et al, 2011).…”

Section: Mode Of Actionmentioning

confidence: 99%

A roadmap for hazard monitoring and risk assessment of marine biotoxins on the basis of chemical and biological test systems

Daneshian

2013

ALTEX

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Summary Aquatic food accounts for over 40% of global animal food products, and the potential contamination with toxins of algal origin -marine biotoxins -poses a health threat for consumers. The gold standards to assess toxins in aquatic food have traditionally been in vivo methodssingle marine biotoxins or combinations of marine biotoxins and intoxication symptomatology as well as monitoring the "success" of controls implemented during fishery and aquaculture production, processing, and retailing. For this reason, a workshop was organized in ermatingen, Switzerland by the Center for Alternatives to Animal testing -europe (CAAteurope) within the framework of the transatlantic think tank for toxicology (t 4 ).In contrast to other food products where hygiene and potential contamination with microbes or mold toxins is the prime issue, the emphasis in aquatic products (shellfish and finfish) is, beyond viral and bacterial contaminations, primarily placed on potential contamination with marine biotoxins owing to the numerous and recurring human intoxications.The gold standards to assess toxins in aquatic food have traditionally been in vivo methods, i.e., the mouse and the rat bioassays. Besides the ethical issues of in vivo bioassays there are specific difficulties, e.g., different exposure route (i.p. versus oral), low inter-species comparability, high intra-species variability, and questionable extrapolation of quantitative risk to humans, thus highlighting the need for the development of more reliable detection and quantification methods. Based on this reasoning, the european Food Safety Authority (eFSA) advocates the use of an analytical method, i.e., LC-MS (Liquid Chromatography coupled Mass Spectrometry), as a substitute for in vivo bioassays for almost all classes of marine toxins. However, although lC-MS is a very sensitive method that has the advantages of being able to detect multiple toxins in a single analysis and being good for confirming the identity of toxins, the quality of quantitative analysis is dependent on the availability of calibration standards. While many new toxin structural analogues have been detected and identified using LC-MS, this technique -as with most other methods -is not good at detecting new toxin types. When new toxin analogues are detected but accurate standards are not yet available, only an approximate quantitation is possible using estimated response factors.For the purpose of risk assessment of food, however, it is imperative to focus on the possible adverse effects, independent of whether they stem from one toxin or a combination of toxins. As toxicity is defined by functional and structural changes of biological systems, the development of a human-relevant in vitro system for appropriate risk assessment of marine biotoxins in seafood stands to reason. Moreover, poor correlation of human intoxications, of acute symptomatology and long-term adverse effects, and of predictive in vitro, in vivo, and analytical detection methodologies of marine biotoxins stems not least from a...

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“…For the detection of known lipophilic toxins such as OA, using CBA-based methods, several protocols have been described for shellfish extraction using solvents such as saline buffer [28], methanol-water [29], and acetone [20,21,30]. In most cases, studies focused on distinguishing between MBA-positive and MBA-negative samples and matrix effects were not taken into consideration (e.g., [20,21]).…”

Section: Toxicity Of Control Shellfish Extractsmentioning

confidence: 99%

“…In most cases, studies focused on distinguishing between MBA-positive and MBA-negative samples and matrix effects were not taken into consideration (e.g., [20,21]). In this study, the same extraction protocol for shellfish digestive glands (an acetone extraction followed by a liquid/liquid partitioning with DCM/water) was performed for the CBA as well as for MBA and LC-MS/MS analysis, to ensure that the unknown lipophilic toxic compounds were recovered.…”

Section: Toxicity Of Control Shellfish Extractsmentioning

confidence: 99%

“…Cell mortality resulting from the impairment of various essential cellular functions is generally considered to provide a relevant approach to detect the largest panel of toxic compounds including phycotoxins [4]. Thus, this parameter was employed to distinguish between mouse bioassay (MBA)-positive and MBA-negative shellfish samples for OA using kidney BGM cells [20] and, for AZAs using HepG2 and bladder ECV-304 cells [21].…”

Section: Introductionmentioning

confidence: 99%

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Collaborative study for the detection of toxic compounds in shellfish extracts using cell-based assays. Part II: application to shellfish extracts spiked with lipophilic marine toxins

et al. 2012

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Successive unexplained shellfish toxicity events have been observed in Arcachon Bay (Atlantic coast, France) since 2005. The positive mouse bioassay (MBA) revealing atypical toxicity did not match the phytoplankton observations or the liquid chromatography-tandem mass spectrometry (LC-MS/MS) investigations used to detect some known lipophilic toxins in shellfish. The use of the three cell lines (Caco2, HepG2, and Neuro2a) allows detection of azaspiracid-1 (AZA1), okadaic acid (OA), or pectenotoxin-2 (PTX2). In this study, we proposed the cell-based assays (CBA) as complementary tools for collecting toxicity data about atypical positive MBA shellfish extracts and tracking their chromatographic fractionation in order to identify toxic compound(s). The present study was intended to investigate the responses of these cell lines to shellfish extracts, which were either control or spiked with AZA1, OA, or PTX2 used as positive controls. Digestive glands of control shellfish were extracted using the procedure of the standard MBA for lipophilic toxins and then tested for their cytotoxic effects in CBA. The same screening strategy previously used with pure lipophilic toxins was conducted for determining the intra- and inter-laboratory variabilities of the responses. Cytotoxicity was induced by control shellfish extracts whatever the cell line used and regardless of the geographical origin of the extracts. Even though the control shellfish extracts demonstrated some toxic effects on the selected cell lines, the extracts spiked with the selected lipophilic toxins were significantly more toxic than the control ones. This study is a crucial step for supporting that cell-based assays can contribute to the detection of the toxic compound(s) responsible for the atypical toxicity observed in Arcachon Bay, and which could also occur at other coastal areas.

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A cytotoxicity assay for the detection and differentiation of two families of shellfish toxins

Cited by 51 publications

References 18 publications

Investigation on neurotoxin of sea snail meat

Investigation on neurotoxin of sea snail meat

A roadmap for hazard monitoring and risk assessment of marine biotoxins on the basis of chemical and biological test systems

Collaborative study for the detection of toxic compounds in shellfish extracts using cell-based assays. Part II: application to shellfish extracts spiked with lipophilic marine toxins

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