Identification of autoclave-resistant Anisakis simplex allergens

Area of Science:

• Foodborne Parasitology
• Clinical Allergy and Immunology
• Food Processing and Safety

Background:

• Anisakis simplex parasites in fish can cause human allergic reactions.
• Allergens may persist after cooking, leading to symptoms from processed fish products.
• Canned fish consumption has been linked to allergic reactions, suggesting heat-stable allergens.

Purpose of the Study:

• To investigate the stability of Anisakis simplex allergens after autoclaving at 121 °C.
• To simulate thermal processing conditions used in industrial canning.
• To identify specific allergens that resist heat treatment and retain allergenic potential.

Main Methods:

• Third-stage Anisakis simplex larvae were autoclaved at 121 °C for varying durations (20, 40, 80 min).
• Crude parasite extracts were analyzed using electrophoresis, immunoblotting, and basophil activation tests.
• Heat-resistant allergens were isolated via HPLC and identified using mass spectrometry.

Main Results:

• Autoclaving reduced protein bands in a time-dependent manner.
• Immunoblotting confirmed the presence of several Anisakis simplex allergens after autoclaving.
• Ani s 1 (14 kDa) and Ani s 4 (9 kDa) were identified as heat-resistant allergens.
• Autoclaved allergens retained their capacity to activate basophils, indicating preserved allergenic function.

Conclusions:

• Significant Anisakis simplex allergens, including Ani s 1 and Ani s 4, are stable under autoclaving conditions.
• These heat-resistant allergens can bind IgE and activate immune cells even after industrial canning-equivalent heat treatment.
• This stability explains allergic reactions in individuals consuming canned fish contaminated with Anisakis simplex.