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Identification of Autoclave-Resistant Anisakis simplex Allergens

AutorCarballeda-Sangiao, Noelia; Olivares, Fabiola; Rodríguez-Mahillo, Ana I.; Careche, Mercedes ; Tejada Yábar, Margarita ; Moneo, Ignacio; González Muñoz, Miguel
Fecha de publicación2014
EditorInternational Association for Food Protection
CitaciónJournal of Food Protection 77: 605- 609 (2014)
ResumenAnisakis simplex is a fish parasite able to induce allergic reactions in humans infected when eating raw or undercooked fish parasitized with viable third-stage larvae. Some authors claim that exposure to nonviable Anisakis material can result in allergic symptoms in previously sensitized patients, indicating that parasite allergens are resistant to the thermal treatments of usual cooking procedures. Furthermore, some patients report symptoms after eating canned fish. The aim of this work was the analysis of parasite allergen stability in heating to 121uC in an autoclave to simulate the thermal process applied to canned fish. Thirdstage larvae were subjected to autoclaving for 20, 40, and 80 min, and parasite crude extracts were analyzed by electrophoresis, immunoblotting, and a flow-cytometric basophil activation test. Allergens resistant to autoclaving were separated by reversedphase high-performance liquid chromatography and identified by ion trap mass spectrometry. Protein analysis by sodium dodecyl sulfate¿polyacrylamide gel electrophoresis showed that autoclaving considerably reduced the number and intensity of identifiable protein bands in a time-dependent manner. Several allergens were detected by immunoblotting with a pool of A. simplex allergic patients¿ sera after autoclaving. Allergens of 9 and 14 kDa resistant to autoclaving were identified as Ani s 4 and Ani s 1 allergens, respectively. Functional analysis showed that allergens retain their capacity to activate basophils even after autoclaving for 80 min. In conclusion, some relevant A. simplex allergens retain their capacity to bind immunoglobulin E and activate basophils after being subjected to autoclaving, which is a method equivalent to that used in industrial canning processes.
URIhttp://hdl.handle.net/10261/108084
DOI10.4315/0362-028X.JFP-13-278
Identificadoresdoi: 10.4315/0362-028X.JFP-13-278
issn: 0362-028X
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