Marine Biotoxins in Shellfish

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Juliana de Smet
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1 Marine Biotoxins in Shellfish Arjen Gerssen, Hester van den Top and Hans van Egmond

2 Background Certain algae can produce toxins These algae can end up in filter feeding shellfish like mussels and oysters Consumption of these contaminated shellfish can cause severe intoxication

3 Background: lipophilic marine toxins Toxins included in this group; Diarrhetic Shellfish Poisoning (DSP) toxins (Okadaic acid and Dinophysistoxins) Azaspiracid Shellfish Poisoning (AZP) toxins Azaspiracids Pectenotoxins Yessotoxins Cyclic imines spirolides

4 Okadaic acid and Dinophysistoxins Produced by (i.e.) Dinophysis acuta Dinophysis acuminata Adverse effects Diarrhoea, abdominal cramps, nausea and vomiting Possible carcinogenic

5 Azaspiracides Produced by Azadinium spinosum Adverse effects Diarrhoea, abdominal cramps, nausea and vomiting Effects comparable to okadaic acid only via another mechanism

6 Yessotoxins Produced by (i.e.) Protoceratium reticulatum Lingulodinium polyedrum Adverse effects Swelling of cardiomyocytes (oedema) Defattening of the liver

7 Pectenotoxins Produced by (i.e.) Dinophysis acuta Dinophysis acuminata Adverse effects Formation of vacuoles in hepatocytes (observed in test animals)

8 Cyclic imines Produced by (i.e.) Alexandrium ostenfeldii Adverse effects Neurotoxic effects in test animals

9 Analysis of DSP en AZP toxins using rat A certain time period after the rat has consumed shellfish material, the consistency of the faeces is checked. Method used in the Netherlands only

10 Analysis of DSP en AZP toxins using rat Advantages No extraction procedure involved No complex expensive equipment Drawbacks Special animal facilities needed Differences in sensitivity (rat to rat) Detectability Non specific for other regulated lipophilic toxins Provide no early warning Animal unfriendly

11 An even worse approach, the mouse test Unreliable Non specific Animal unfriendly And all the drawbacks of the rat bioassay On a EU level mice are annually used for shellfish monitoring!!!!

12 Alternative for rats and mice Chemical testing (based on LC-MS/MS) Advantage Sensitivity Specificity Obstacles Relative expensive machinery Validated method needed Legislation (was) not ready for alternative

13 Chemical alternative Method validation (determination performance characteristics) Repeatability Reproducibility Sensitivity Ruggedness Within lab (RIKILT) Excellent performance Between lab (13 international participants) Excellent performance

14 Interlaboratory validation study in participants - 12 EU and 1 USA Performed according to international guidelines (AOAC guidelines appendix D) Excellent performance for various toxins in various shellfish materials Method can be used for official control as EU legislation has changed

15 New EU legislation in 2011 Alternative method available instead of using rats and mice New legislation will be in place 1 st of July 2011 The Netherlands will use the alternative method developed at RIKILT For other EU countries: 31/12/2014 will be the final day that an animal test is allowed for the detection of regulated toxins

16 RIKILT Institute of Food Safety - Wageningen UR, Arjen.Gerssen@wur.nl

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