Authentication of seafood species on the ASFIS list (FAO) by in-silico evaluation of primers for metabarcoding

Given the remarkable biodiversity among the seafood species entering the food supply chain, authentication is a crucial component in ensuring transparent trade. DNA metabarcoding stands out as the leading methodology for determining species composition within complex food matrices. Nevertheless, the selection of primer pairs is critical, as it may introduce biases into species identification. In this investigation, we evaluated previously described primers for their in-silico efficacy in identifying both edible marine species on the ASFIS (FAO) list and the main terrestrial animal species frequently incorporated as ingredients in processed seafood products. Selected primers covered the Cytochrome c Oxidase subunit I (COI), the Cytochrome b (cytb), the 16S rRNA (16S) and the 12S rRNA (12S) mitochondrial regions. Results showed that, out of 53 pairs analysed, one primer pair targeting the COI region obtained the best performance in terms of taxonomic coverage for both ASFIS and terrestrial species. Notably, each primer pair showed a unique taxonomic profile, suggesting their potential for addressing various seafood authentication issues as the detection of novel food ingredients (e.g., jellyfish, red algae, insects) or endangered species. Overall, these outcomes deliver valuable insights for selecting metabarcoding primers to improve the traceability of seafood products and contribute to the development of an innovative Food Safety Management System.