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Towards the standardisation of the neuroblastoma (neuro-2a) cell-based assay for ciguatoxin-like toxicity detection in fish: application to fish caught in the Canary Islands.

Abstract

The ouabain/veratridine-dependent neuroblastoma (neuro-2a) cell-based assay (CBA) was applied for the determination of the presence of ciguatoxin (CTX)-like compounds in ciguatera-suspected fish samples caught in the Canary Islands. In order to avoid matrix interferences the maximal concentration of wet weight fish tissue exposed to the neuro-2a cells was set at 20 mg tissue equivalent (TE) ml(-1) according to the sample preparation procedure applied. In the present study, the limit of quantification (LOQ) of CTX1B equivalents in fish extract was set at the limit of detection (LOD), being defined as the concentration of CTX1B equivalents inhibiting 20% cell viability (IC(20)). The LOQ was estimated as 0.0096 ng CTX1B eq.g TE(-1) with 23-31% variability between experiments. These values were deemed sufficient even though quantification given at the IC(50) (the concentration of CTX1B equivalents inhibiting 50% cell viability) is more accurate with a variability of 17-19% between experiments. Among the 13 fish samples tested, four fish samples were toxic to the neuro-2a cells with estimations of the content in CTX1B g(-1) of TE ranging from 0.058 (± 0.012) to 6.23 (± 0.713) ng CTX1B eq.g TE(-1). The high sensitivity and specificity of the assay for CTX1B confirmed its suitability as a screening tool of CTX-like compounds in fish extracts at levels that may cause ciguatera fish poisoning. Species identification of fish samples by DNA sequence analysis was conducted in order to confirm tentatively the identity of ciguatera risk species and it revealed some evidence of inadvertent misidentification. Results presented in this study are a contribution to the standardisation of the neuro-2a CBA and to the risk analysis for ciguatera in the Canary Islands.

Authors+Show Affiliations

IRTA, Ctra. Poble Nou, km. 5.5, E-43540 Sant Carles de Ràpita, Spain.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

22394180

Citation

Caillaud, A, et al. "Towards the Standardisation of the Neuroblastoma (neuro-2a) Cell-based Assay for Ciguatoxin-like Toxicity Detection in Fish: Application to Fish Caught in the Canary Islands." Food Additives & Contaminants. Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment, vol. 29, no. 6, 2012, pp. 1000-10.
Caillaud A, Eixarch H, de la Iglesia P, et al. Towards the standardisation of the neuroblastoma (neuro-2a) cell-based assay for ciguatoxin-like toxicity detection in fish: application to fish caught in the Canary Islands. Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2012;29(6):1000-10.
Caillaud, A., Eixarch, H., de la Iglesia, P., Rodriguez, M., Dominguez, L., Andree, K. B., & Diogène, J. (2012). Towards the standardisation of the neuroblastoma (neuro-2a) cell-based assay for ciguatoxin-like toxicity detection in fish: application to fish caught in the Canary Islands. Food Additives & Contaminants. Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment, 29(6), 1000-10. https://doi.org/10.1080/19440049.2012.660707
Caillaud A, et al. Towards the Standardisation of the Neuroblastoma (neuro-2a) Cell-based Assay for Ciguatoxin-like Toxicity Detection in Fish: Application to Fish Caught in the Canary Islands. Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2012;29(6):1000-10. PubMed PMID: 22394180.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Towards the standardisation of the neuroblastoma (neuro-2a) cell-based assay for ciguatoxin-like toxicity detection in fish: application to fish caught in the Canary Islands. AU - Caillaud,A, AU - Eixarch,H, AU - de la Iglesia,P, AU - Rodriguez,M, AU - Dominguez,L, AU - Andree,K B, AU - Diogène,J, Y1 - 2012/03/07/ PY - 2012/3/8/entrez PY - 2012/3/8/pubmed PY - 2012/8/17/medline SP - 1000 EP - 10 JF - Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment JO - Food Addit Contam Part A Chem Anal Control Expo Risk Assess VL - 29 IS - 6 N2 - The ouabain/veratridine-dependent neuroblastoma (neuro-2a) cell-based assay (CBA) was applied for the determination of the presence of ciguatoxin (CTX)-like compounds in ciguatera-suspected fish samples caught in the Canary Islands. In order to avoid matrix interferences the maximal concentration of wet weight fish tissue exposed to the neuro-2a cells was set at 20 mg tissue equivalent (TE) ml(-1) according to the sample preparation procedure applied. In the present study, the limit of quantification (LOQ) of CTX1B equivalents in fish extract was set at the limit of detection (LOD), being defined as the concentration of CTX1B equivalents inhibiting 20% cell viability (IC(20)). The LOQ was estimated as 0.0096 ng CTX1B eq.g TE(-1) with 23-31% variability between experiments. These values were deemed sufficient even though quantification given at the IC(50) (the concentration of CTX1B equivalents inhibiting 50% cell viability) is more accurate with a variability of 17-19% between experiments. Among the 13 fish samples tested, four fish samples were toxic to the neuro-2a cells with estimations of the content in CTX1B g(-1) of TE ranging from 0.058 (± 0.012) to 6.23 (± 0.713) ng CTX1B eq.g TE(-1). The high sensitivity and specificity of the assay for CTX1B confirmed its suitability as a screening tool of CTX-like compounds in fish extracts at levels that may cause ciguatera fish poisoning. Species identification of fish samples by DNA sequence analysis was conducted in order to confirm tentatively the identity of ciguatera risk species and it revealed some evidence of inadvertent misidentification. Results presented in this study are a contribution to the standardisation of the neuro-2a CBA and to the risk analysis for ciguatera in the Canary Islands. SN - 1944-0057 UR - https://www.unboundmedicine.com/medline/citation/22394180/Towards_the_standardisation_of_the_neuroblastoma__neuro_2a__cell_based_assay_for_ciguatoxin_like_toxicity_detection_in_fish:_application_to_fish_caught_in_the_Canary_Islands_ L2 - https://www.tandfonline.com/doi/full/10.1080/19440049.2012.660707 DB - PRIME DP - Unbound Medicine ER -