Molecular Identification of Anisakis larvae (Nematode: Anisakidae) from marine fishes collected from three Korean sea waters

Abstract

Anisakis infects humans who eat raw seafood such as squid, mackerel, and conger, which is considered as one of the parasitic diseases in Korea. Molecular study for Anisakis L3 larvae was performed to identify the species of Anisakis. Molecular analysis for Anisakis L3 larvae was performed to identify the species of Anisakis and by using the DNA alignment tool to identify the DNA sequence relation. Total 98 Anisakis type I L3 larvae were collected from 63 fishes among three kinds of fish species (mackerel (Scomberomorus japonicus), anago (Conger myriaster), and yellow croaker (Larimichthys polyactis)) that came from different location near sea areas (Namhae-gun, Uljin-gun, Daecheon-si, Pohang-si and Seoul). DNA were extracted from Anisakis and ITS gene (forward primer: Nc5 (5’-GTAGGTGAACCTGCGGAAGGATCATT-3’); reverse primer: Nc2 (5’-TTAGTTTCTTTTCCTCCGCT-3’)) was amplified using PCR and sequenced. By using some method that considered for the species identification strategies, such as: Sequence analysing (Anisakis pegreffi (GGCAGAGTC) => C/C allele and Anisakis simplex (GGCAGAGTT) => T/T allele) and the strategy to find the rest total of unidentified in the first strategy by using nucleotide identity percentage: A. pegreffi (AB277823), A. simplex (AB277822), P. decipiens (AB277824) and H. aduncum (AB277826). It was found that Anisakis pegreffi was the most predominant Anisakis species from all fish species and in all Korean locations with 67 as a result of the total. And in the result of nucleotide identity percentage for analyzed the rest 31 samples that not identified during the sequence procedure, based on matched result, 1 were more matched to A. simplex, 25 to A. pegreffi and 5 to H. aduncum.