Validation of the TrichinEasy® digestion system for the detection of Anisakidae larvae in fish products

Gaetano Cammilleri; Michele Chetta; Antonella Costa; Stefania Graci; Rosaria Collura; Maria Drussilla Buscemi; Maria Cusimano; Angelina Alongi; Deborah Principato; Giuseppe Giangrosso; Antonio Vella; Vincenzo Ferrantelli

doi:10.1515/ap-2016-0048

Article

Validation of the TrichinEasy® digestion system for the detection of Anisakidae larvae in fish products

Gaetano Cammilleri , Michele Chetta , Antonella Costa , Stefania Graci , Rosaria Collura , Maria Drussilla Buscemi , Maria Cusimano , Angelina Alongi , Deborah Principato , Giuseppe Giangrosso , Antonio Vella and Vincenzo Ferrantelli

Published/Copyright: March 30, 2016

Published by

Become an author with De Gruyter Brill

Author Information

From the journal Acta Parasitologica Volume 61 Issue 2

Abstract

Anisakis and other parasites belonging to the Anisakidae family are organisms of interest for human health, because of their high zoonotic potential. Parasites belonging to this family can cause Anisakiasis, a parasitological disease caused by the ingestion of raw, infested fish products. Furthermore, evidence from the EFSA (European Food Safety Authority; EFSA 2010) has highlighted the allergological potential of nematodes belonging to the Anisakis genre. The detection and identification of Anisakidae larvae in fish products requires an initial visual inspection of the fish sample, as well as other techniques such as candling, UV illumination and artificial digestion. The digestion method consists of the simulation of digestive mechanics, which is made possible by the utilization of HCl and pepsin, according to EC Regulation 2075/2005. In this study, a new Anisakidae larvae detection method using a mechanical digestion system called Trichineasy^® was developed. A total of 142 fish samples, belonging to 14 different species, were examined to validate the method. A reaction mixture with 100 g of sample, 10 g of pepsin (1:10000 NF) and 50 ml of 10% HCl at 36 ± 1°C for 20 minutes was evaluated to be the best condition for the digestion of fish samples. These parameters have also allowed the detection of viable larvae after digestion. The results confirm this instrumentation as a valuable and safe tool for the detection of Anisakidae larvae in fishery products.

Keywords: Anisakidae nematodes; artificial digestion; instrument validation

References

Abollo, E., D’Amelio, S., Pascual, S. 2001. Fitness of the marine parasitic nematode Anisakis simplex s. str. in temperate waters of the NE Atlantic. Diseases of Aquatic Organisms 45, 131–139. DOI:10.3354/dao04513110.3354/dao045131Search in Google Scholar PubMed

Alonso A., Daschner A., Moreno-Ancillo A. 1997. Anaphylaxis with Anisakis simplex in the gastric mucosa. The New England Journal of Medicine 337, 350–351. DOI: 10.1056/NEJM199 70731337051810.1056/NEJM199707313370518Search in Google Scholar PubMed

Alonso-Gomez A., Moreno-Ancillo A., Lopez-Serrano M.C., Suarez-de-Parga J.M., Daschner A. Caballero M.T., Barranco P., Cabanas R. 2004. Anisakis simplex only provokes allergic symptoms when the worm parasitises the gastrointestinal tract. Parasitology Research 93, 378–384. DOI: 10.1007/s00436-004-1085-910.1007/s00436-004-1085-9Search in Google Scholar PubMed

Aly M., Safwat M., Mohamed M. 2000. Anisakis and fish induced urticaria. Journal of the Egyptian Society of Parasitology 30, 601–606Search in Google Scholar

Anandon A.M., Romaris F., Escalante M., Rodriguez E., Garate T., Cuellar C., Ubeira F.M. 2009. The Anisakis simplex Ani s 7 major allergen as an indicator of true Anisakis infections. Clinical & Experimental Immunology 156, 471–47810.1111/j.1365-2249.2009.03919.xSearch in Google Scholar PubMed PubMed Central

COMMISSION REGULATION (EC) No 2074/2005 of 5 December 2005 laying down implementing measures for certain products under Regulation (EC) No 853/2004 of the European Parliament and of the Council and for the organisation of official controls under Regulation (EC) No 854/2004 of the European Parliament and of the Council and Regulation (EC) No 882/2004 of the European Parliament and of the Council, derogating from Regulation (EC) No 852/2004 of the European Parliament and of the Council and amending Regulations (EC) No 853/2004 and (EC) No 854/2004Search in Google Scholar

COMMISSION REGULATION (EC) No 2075/2005 of 5 December 2005 laying down specific rules on official controls for Trichinella in meatSearch in Google Scholar

EFSA Scientific Opinion on risk assessment of parasites in fishery products. EFSA journal 2010; 8, 1543.10.2903/j.efsa.2010.1543Search in Google Scholar

Ferrantelli V., Cicero A., Costa A., Alongi A., Palumbo P., Graci S., Giangrosso G. 2014. Anisakidae in fishing products sold in Sicily. Italian Journal of Food Safety 3, 47–48. DOI: 10.4081/ijfs.2014.171910.4081/ijfs.2014.1719Search in Google Scholar PubMed PubMed Central

Ferrantelli V., Costa A., Graci S., Buscemi M. D., Giangrosso G., Porcarello C., Palumbo S., Cammilleri G. 2015. Anisakid nematodes as possible markers to trace fish products. Italian Journal of Food Safety 4, 49–53 DOI: 10.4081/ijfs.2015.409010.4081/ijfs.2015.4090Search in Google Scholar PubMed PubMed Central

Karl H., Levsen A. 2011. Occurrence and distribution of anisakid nematodes in Grey gurnard (Eutrigla gurnardus L.) from the North Sea. Food Control 22, 1634–163810.1016/j.foodcont.2011.03.021Search in Google Scholar

Levsen A., Karl H. 2014. Anisakis simplex (s.l.) in Grey gurnard (Eutrigla gurnardus) from the North Sea: Food safety considerations in relation to fishing ground and distribution in the flesh. Food Control 36, 15–19. DOI:10.1016/j.foodcont. 2013.07.00610.1016/j.foodcont.2013.07.006Search in Google Scholar

Levsen, A., Lunestad, B. T., Berland B. 2005. Low Detection Efficiency of Candling as a Commonly Recommended Inspection Method for Nematode Larvae in the Flesh of Pelagic Fish. Journal of Food Protection 68, 828–83210.4315/0362-028X-68.4.828Search in Google Scholar

Llarena-Reino M., Pineiro C., Antonio J., Outerino L., Carlos Vello, González A. F., Pascual S. 2013. Optimization of the pepsin digestion method for anisakids inspection in the fishing industry. Veterinary Parasitology 191, 276–28310.1016/j.vetpar.2012.09.015Search in Google Scholar

Mattiucci S., Cipriani P., Webb S. C., Paoletti M., Marcer F., Bellisario B., Gibson D. I., and Nascetti G. 2014. Genetic and Morphological Approaches Distinguish the Three Sibling Species of the Anisakis simplex Species Complex, with a Species Designation as Anisakis berlandi n. sp. for A. simplex sp. C (Nematoda: Anisakidae). Journal of Parasitology 100, 199–21410.1645/12-120.1Search in Google Scholar

Mattiucci S., Nascetti G. 2008. Chapter 2 Advances and Trends in the Molecular Systematics of Anisakid Nematodes, with Implications for their Evolutionary Ecology and Host-Parasite Co-evolutionary Processes. Advances in Parasitology 66, 47– 148 DOI: 10.1016/S0065-308X(08)00202-910.1016/S0065-308X(08)00202-9Search in Google Scholar

Sakanari J.A., McKerrow J.H. 1989. Anisakiasis. Clinical Microapy Reviews 2, 278–28410.1128/CMR.2.3.278Search in Google Scholar PubMed PubMed Central

Received: 2015-8-12

Revised: 2015-11-4

Accepted: 2015-12-14

Published Online: 2016-3-30

Published in Print: 2016-6-1

You are currently not able to access this content.

Articles in the same Issue

https://doi.org/10.1515/ap-2016-0048

Keywords for this article

Anisakidae nematodes; artificial digestion; instrument validation