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A method to detect the parasitic nematodes from the family Anisakidae, in Sardina pilchardus, using specific primers of 18 S DNA gene

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Abstract

Anisakiasis is the infestation of man by the third larval stage of the Anisakidae family parasites, caused by the consumption of raw or undercooked seafood. The hosts of the third larvae parasite are marine fishes, among which Sardina pilchardus is of high commercial interest in many countries. So, the detection of anisakid larvae in fisheries products (fresh and processed) is of great interest to a rapid and feasible diagnosis of parasitism. Currently, there are different types of parasite detection methods but they are all difficult and tedious. The aim of this work was to detect, using DNA-based techniques, the presence of anisakid larvae in the entire fish host. Three pairs of primers were designed in the 18 S DNA gene taking into account the maximum matching between the anisakid species sequences and the minimum matching concerning for both the fish species and some other groups of organisms that parasitize fish (non-anisakid nematodes, trematodes, and cestodes). Experimental mixtures of parasite and fish host were made and the sensitivity of anisakid DNA detection tested by PCR. All the above primers and two others combinations of them were specific for the nematode group within the studied species, and the sensitivity of detection was on the order of 1 part of nematode to 100 000 parts of fish (anisakid weight:fish sample fresh weight = 1:≈100 000).

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Acknowledgements

We wish to thank René Galzin and Joseph Jourdane who kindly provided all the laboratory support for the work. We acknowledge the technical assistance on equipment and services managed by Jean François Allienne. We also thank Paula Ramos from IPIMAR and Stephanie Leroy, Jérome Boissier, and Andrea Simkova from the Laboratoire de Biologie Animale for gifts of nematode and trematode samples. Finally, we are grateful to Eric Taylor for his advices on the English correction of this manuscript. The work was supported by the grant SFRH/BSAB/223/2001 from POCTI (Formar e Qualificar–Medida 1.1) from FCT

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Authors and Affiliations

  1. INIAP/Instituto de Investigação das Pescas e do Mar (IPIMAR), Departamento de Inovação Tecnológica e Valorização dos Produtos da Pesca Av., Brasilia, Lisboa, Brazil

    Ana Teia Santos

  2. Laboratoire de Biologie Animale, UMR 5555 CNRS, Université de Perpignan, 52 avenue de Villeneuve, 66860, Perpignan Cedex, France

    Pierre Sasal & Olivier Verneau

  3. Laboratoire d’Ichtyoécologie Tropicale et Méditerranéenne, École Pratique des Hautes Études–ESA 8146 CNRS, Université de Perpignan, 52 avenue de Villeneuve, 66860, Perpignan Cedex, France

    Philippe Lenfant

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  1. Ana Teia Santos
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  2. Pierre Sasal
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  3. Olivier Verneau
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  4. Philippe Lenfant
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Correspondence to Ana Teia Santos.

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Santos, A.T., Sasal, P., Verneau, O. et al. A method to detect the parasitic nematodes from the family Anisakidae, in Sardina pilchardus, using specific primers of 18 S DNA gene. Eur Food Res Technol 222, 71–77 (2006). https://doi.org/10.1007/s00217-005-0052-8

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  • DOI: https://doi.org/10.1007/s00217-005-0052-8

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