Overview
EN ISO 23036-2:2021 (ISO 23036-2:2021) specifies the artificial digestion method for detection of Anisakidae L3 larvae in fish and fishery products. This CEN-endorsed standard covers enzymatic (pepsin‑HCl) digestion of fish muscle and related sample types, followed by filtration and microscopic examination to detect and quantify third‑stage Anisakidae larvae (genera such as Anisakis, Contracaecum, Pseudoterranova). It applies to fresh and frozen fish, lightly processed products (marinated, salted, smoked) and to visceral organs as a confirmatory method supporting visual inspection schemes.
Key topics and requirements
- Scope and applicability: Detection and quantification of Anisakidae L3 in marine and anadromous fishes; suitability for fresh, frozen, and lightly processed products; viability assessment only for samples never frozen before processing.
- Principle: Enzymatic digestion (pepsin‑HCl) to release larvae from muscle tissue, followed by filtration and microscopy to recover and count larvae.
- Sample handling: Guidance on sample size, preparation and collection (see Annex A and laboratory worksheets).
- Procedure components: Preparation of digest fluid, digestion step, filtration of digest fluid, microscopic examination and verification of findings.
- Results and reporting: Expression of infestation levels, test report content and performance characteristics of the method.
- Quality assurance: Laboratory quality control, documentation and conformity with general microbiology requirements (reference to ISO 7218).
- Limitations: The method does not identify species or genotype; final identification requires morphological and/or molecular methods.
Applications
- Routine food safety testing in seafood processing and inspection to quantify Anisakidae infestation levels.
- Confirmatory testing following visual inspection of fish and fishery products.
- Determination of larval viability for fresh or lightly processed (never frozen) products to assess consumer risk.
- Use in reference and diagnostic laboratories for surveillance, outbreak investigation and research on anisakidosis prevalence.
Who should use this standard
- Regulatory authorities and official control laboratories involved in seafood safety.
- Food industry quality and HACCP teams in fisheries, processors and cold‑chain operators.
- Public health and research laboratories performing parasitology diagnostics and surveillance.
- Certification bodies and auditors assessing compliance with seafood safety requirements.
Related standards
- ISO 7218 (General requirements for microbiological examinations)
- Other parts of the ISO 23036 series (see ISO website for the full list)
- Complementary detection approaches (e.g., UV‑press method) for comprehensive inspection programs
Keywords: ISO 23036-2:2021, EN ISO 23036-2:2021, Anisakidae L3 larvae, artificial digestion method, fish and fishery products, pepsin‑HCl digestion, seafood safety, larval detection, microscopy, filtration.
