Molecular Identification of Anisakid Nematodes in Imported Marine Fish and Fishery Products in Serbia

Anisakid nematodes are important foodborne hazards in marine fish, but molecular data from imported seafood examined within the Serbian official control system remain scarce. This study characterized anisakid and related ascaridoid molecular profiles in parasite material recovered during official examination of imported marine fish and fishery products. Seventy-four selected molecular test records were analysed by multiplex PCR targeting the internal transcribed spacer region of ribosomal DNA. Specific amplification was obtained in 55 records (74.3%). The predominant result was an Anisakis simplex sensu lato/A. pegreffii hybrid-like multiplex-PCR banding profile (39 records), followed by A. pegreffii (8), Contracaecum osculatum (6), and Anisakis typica (2); Anisakis taxa accounted for 89.1% of positive records. Hake (Merluccius spp.) predominated among records with known fish identity, although record-level positivity was similar in hake and mackerel (Scomber spp.). Spain accounted for most records with a known origin, while positivity and profile distributions were descriptively comparable with other known origins. Multiplex-PCR banding profiles provide a useful complement to routine parasitological examination and baseline descriptive information for surveillance of anisakid-related hazards in imported seafood in Serbia; however, the assay alone cannot confirm hybrid genotypes or parasite viability. The findings describe selected laboratory molecular-test records and do not constitute estimates of anisakid prevalence in imported fish.