Marinated fish treatment using low pH to enlarge the storage life of fish as in anchovies in vinegar, does not kill Anisakis simplex larvae infesting fish muscle. To kill the larvae it is compulsory in many countries to freeze fish intended to be marinated raw, which prevents the consumer to be infested with the live larvae. However, it is not known if A. simplex antigens are released to the media after freezing and vinegar processing, which may cause allergic reaction to A. simplex sensitized consumers. Anchovy fillets were artificially infested with A. simplex L3, treated with a vinegar solution and chilled stored for 10 days. Infested frozen-thawed fillets were treated and stored in the same conditions. Viability of the larvae, SEM, immunoblotting and immunohistochemistry were performed on the treated fillets before and after pepsin treatment. Viability of the larvae was detected only in the chilled fillets; however, A. simplex antigens were detected in the chilled and in the frozen-thawed fillets even after pepsin treatment. This suggests that the consumption of anchovies in vinegar may be a potential hazard when ingested by sensitized consumers, even if freezing kills the larvae.
Anonymous (1995). Isolation and identification of Anisakid round-worm larvae in fish. Bureau of Microbial Hazards. Food Directorate. Laboratory Procedure ExFLP-1. Health Canada. Health products and food branch, Ottawa . Available from: http://www.hc-sc.gc.ca/fn-an/res-rech/analy-meth/microbio/volume5/opflp_02-eng.php (accessed February 25, 2009).
2.
Aoac (1995). Official Methods of Analysis, 16th edn, Arlington, Va: Association of Official Analytical Chemists.
3.
Caballero M.L. and Moneo I. (2004). Several allergens from Anisakis simplex are highly resistant to heat and pepsin treatments. Parasitology Research93: 248-251.
4.
Dzik J.M. (2006). Molecules released by helminth parasites involved in host colonization. Acta Biochimica Polonica53(1): 33-64.
5.
European Union (1991). Council Directive 91/493/EEC laying down the health conditions for the production and the placing on the market of fishery products. OJ L 268 24.9.1991.
6.
European Union (2004). Corrigendum to Regulation (EC) No 853/2004 of the European Parliament and of the Council of 29 April 2004 laying down specific hygiene rules for food of animal origin . OJ L 139 Bull 30.4.2004.
7.
FDA (2001). Processing parameters needed to control pathogens in cold smoked fish. In: Potential Hazards in Cold-smoked Fish: Parasites, Rockville, MD: US Food and Drug Administration, Center for Food Safety and Applied Nutrition .
8.
Garcia M., Moneo I., Audicana M.T., del Pozo M.D., Muñoz D., Fernandez E., Diez J., Etxenagusia M.A. and Ansotegui I.J. (1997). The use of IgE immunoblotting as a diagnostic tool in Anisakis simplex allergy. Journal of Allergy and Clinical Immunology99(4): 497-501.
9.
Hitchcock D.J. (1950). Parasitological study on the Eskimos in the Bethel area of Alaska. Journal of Parasitology36: 232-234.
10.
Karl H., Roepstorff A., Huss H.H. and Bloemsma B. (1995). Survival of Anisakis larvae in marinated herring fillets. International Journal of Food Science and Technology29: 661-670.
11.
Kasuya S. and Koga K. (1992). Significance of detection of specific IgE in Anisakis-related diseases. Arerugi41: 106-110.
12.
Kirstein F., Nieuwenhuizen N.E., Worsnell G., Brombacher F., Mattiucci S. and Lopata A.L. (2007). Different routes of sensitization to Anisakis pegreffii in a mouse model of allergic asthma. Parassitologia49(2): 222.
Knudsen L., Reimers K., Berner L. and Jensen N.C. (1985). A modification of Bligh and Dyer's oil extraction method reducing chloroform vapour outlet. In: 15th Western European Fish Technologists' Association (WEFTA) Meeting, Hamburg, Germany.
15.
Marrone R., Damiano S., Panzardi M., Palma G., Mattiucci S. and Anastasio A. (2007). Occurrence of Anisakis pegreffii larvae in anchovies (Engraulis encrasicolus) caught off Naples Gulf (Italy). Parassitologia49(2): 225.
16.
Moneo I., Caballero M.L. and Jimenez S. (2000). Inmunodetección de IgE específica (immunoblotting) en el estudio de prevalencia de sensibilización a Anisakis simplex en España . Alergología e Inmunología Clínica15: 255-261.
17.
Moneo I. and Caballero M.L. (2002). Anisakis simplex larvae release allergens during a short incubation in diluted acid which can be used for clinical diagnosis . Alergología e Inmunología Clínica17: 201-207.
18.
Morris S.R. and Sakanari J.A. (1994). Characterization of the serine protease and serine protease inhibitor from the tissue-penetrating nematode Anisakis simplex. Journal of Biological Chemistry269: 27650-27656.
19.
Ng K.S., Petersen J.F.W., Cherney M.M., Garen C., James M.N.G., Zalatoris J.L., Rao-Naik C., Dunn B.M., Martzen M.R. and Peanasky R.J. (1999). Structure determination of Ascaris pepsin inhibitor-3 by MAD phasing. Available from: http:// www.aps.anl.gov/News/Reports/1999/ngk1.pdf (accessed August 10, 2007).
20.
Pichler W.J. (1999). Anisakiasis: immunity, allergy or both? Lessons on the natural role of immunoglobulin E from a nematode infestation. Clinical and Experimental Allergy29: 1161-1163.
21.
Rodriguez-Mahillo A.I., Gonzalez-Muñoz M., Gomez-Aguado F., Rodriguez-Perez R., Corcuera M.T., Caballero M.L. and Moneo I. (2007). Cloning and characterisation of the Anisakis simplex allergen Ani s 4 as a cysteine-protease inhibitor. International Journal for Parasitology37(8-9): 907-917.
22.
Sanchez-Monsalvez I., De Armas-Serra C., Martínez J., Dorado M., Sánchez A. and Rodríguez-Caabeiro F. (2005). A new procedure for marinating fresh anchovies and ensuring the rapid destruction of Anisakis Larvae. Journal of Food Protection68(5): 1066-1072.
23.
Solas M.T., García M.L., Rodriguez-Mahillo A.I., González-Munoz A.I., De las Heras C. and Tejada M. (2008). Anisakis antigens detected in fish muscle infested with Anisakis simplex L3. Journal of Food Protection71(6): 1273-1276.
24.
Tejada M., Careche M., Torrejon P., Del Mazo M.L., Solas M.T., García M.L. and Barba C. (1996). Protein extracts and aggregates forming in minced cod (Gadus morhua) during frozen storage. Journal of Agricultural and Food Chemistry44: 3308-3314.
25.
Tejada M., Solas M.T., Navas A. and Mendizábal A. (2006a). Effect of freezing and different heat treatments on Anisakis larvae: preliminary study, In: Luten J.B., Jacobsen C., Bekaert K., Sæbø A. and Oehlenschläger J. (eds), Seafood Research from Fish to Dish, Quality, Safety and Processing of Wild and Farmed Fish, Wageningen: Academic Publishers , pp. 309-316.
26.
Tejada M., Solas M.T., Navas A. and Mendizábal A. (2006b). Scanning electron microscopy of Anisakis larvae following different treatments. Journal of Food Protection69(6): 1379-1387.
