Development of tools for product control and human risk evaluation in fishery value chains

The project intends to address the needs of both national and international food safety authorities and the fish processing and exporting industry of analytical tools to document the presence or absence, levels and possible effects of contamination with Anisakis simplex larvae or molecular traces thereof, in various fishery value chains. The tools will be based on visual, DNA-based PCR, immunology or proteomics and will cover different needs of detection (from yes/no to quantification of protein fragments) in fish, foods and feeds, and provide data needed in future risk assessments. The project is a cooperation between the Norwegian Veterinary Institute, the National Institute of Nutrition and Seafood Research (NIFES), Haukeland University Hospital Bergen, the Institute for Molecular Bioscience (IMBV) at the University of Oslo, and the Clinical Research & Development Mass Spectrometry Core Facility at the University of Colorado Denver.

Anisakis simplex on cod filet

The third stage larvae of the parasitic nematode Anisakis simplex (herring or whale worm) commonly occur in virtually all commercially exploited fish stocks in temperate seas. The larvae are mainly situated in the visceral cavity; however, a minor proportion may sometimes migrate deeply into the flesh. Currently, Anisakis is detected in foods by visual inspection or PCR.

Anisakis simplex in fish represents a significant consumer health risk:

  • Anisakiasis, human infection with live larvae, occurs by consuming raw or undercooked fish.
  • Allergy is elicited by Anisakis proteins (11 known allergens) in processed seafood.

A quantitative sandwich ELISA for the detection of Anisakis simplex protein in seafood
We recently developed and validated a sensitive, specific and quantitative sandwich ELISA for the detection of Anisakis proteins in seafood products. A pilot survey on processed Norwegian seafoods was performed. The ELISA was reliable, showing high selectivity, sufficient recovery and good sensitivity with an LOD at 0.3 mg/kg Anisakis simplex protein in processed seafoods. The pilot survey revealed low levels of Anisakis simplex protein in some mackerel products.

See publication of our quantitative sandwich ELISA detecting Anisakis protein in seafood in European Food Research and Technologies

The Norwegian Food Safety Authority is currently planning to perform a survey on raw fish products (Sushi).

Anisakis simplex proteins – proteomics
Proteins are studied and identified by gel electrophoresis, Western blot and LC/MS-MS-analysis.

Publications

  • Werner MT, Fæste CK, Levsen A, Egaas E (2011). A quantitative Sandwich ELISA for the detection of Anisakis simplex protein in seafood. Eur. Food Res. Technol. 232:157-166.
  • Levsen A , Faeste CK, Lin AH , Van Do T, Florvaag E,  Egaas, E  (2013). Consumer health risk posed by Anisakis simplex in Northern Europe – an update. Trop Med Int Health 18:49.
  • Fæste CK, Jonscher KR, Dooper MMBW, Egge-Jacobsen WM, Moen A, Daschner A, Egaas E, Christians U (2014). Characterization of potential novel allergens in the fish parasite Anisakis simplex.  EuPa Open Proteomics J. 4:140-155.
  • Fæste CK, Plassen C, Løvberg KE, Moen A, Egaas E (2015). Detection of proteins from the fish parasite Anisakis simplex in Norwegian farmed salmon and processed fish products. Food Analyt Meth. 8:1390-1402.
  • Fæste CK, Levsen A, Lin AH, Larsen N, Plassen C, Moen A, Van Do T, Egaas E (2015). Fish feed as source of potentially allergenic peptides from the fish parasite Anisakis simplex (sl). Anim Feed Sci Technol 202:52-61.

Presentations

  • Werner MT, Fæste CK, Levsen A, Egaas E. ELISA for the determination of the herring worm Anisakis simplex in foods. 124th AOAC Annual Meeting and Exposition. 26.9.-29.9. 2010. Orlando, FL, USA.
  • Dooper MMBW, Myrset HR, Fæste CK, Egaas E, Van Do T, Florvaag E. IgE-mediated responses towards fish parasite Anisakis, Crab and house dust mite in Norwegian shrimp allergic individuals. XXII World Allergy Congress; 4.-8.12., 2011, Cancun, Mexico.
  • Fæste CK, Dooper MMBW,  Daschner A, Egge-Jacobsen W, Egaas E. Identification of novel allergens in the fish parasite Anisakis simplex. XXII World Allergy Congress; 4.-8.12., 2011, Cancun, Mexico.
  • Egaas E, Fæste CK, Levsen A. Kveis i norsk oppdrettsfisk og fiskefôr. Havbruk 2012; 18.4.2012, Stavanger, Norway
  • Fæste CK, Dooper MMBW, Egge-Jacobsen WM, Daschner A, Egaas E, Jonscher KR, Christians U. Potential new allergens of the fish parasite Anisakis simplex. 126th AOAC Annual Meeting and Exposition. 30.9.-3.10. 2012. Las Vegas, NV, USA.
  • Fæste CK, Egaas E, Larsen N, Plassen C, Fjeldstad L, Løvberg KE, Levsen A. Surveying the fish parasite Anisakis simplex in Norwegian fish products. 126th AOAC Annual Meeting and Exposition. 30.9.-3.10. 2012. Las Vegas, NV, USA.
  • Faeste CK. Development of liquid chromatography-tandem mass spectrometry methods for the quantitation of Anisakis simplex proteins in fish. Current Trends in Mass Spectrometry. 20.-22.7.2016, Chicago, IL, USA.