Identification and genetic characterization of Anisakis larvae from marine fishes in the South China Sea using an electrophoretic-guided approach

Abstract

From 35 species of marine fishes (n = 327) from the South China Sea, 237 nematode larvae were collected and identified morphologically as Anisakis. Genomic DNA was isolated from each larva and subjected to PCR-based RFLP and targeted sequencing of a nuclear ribosomal DNA region between the 3′-end of the small subunit and 5′-end of the large subunit of the rRNA genes (= internal transcribed spacers, ITS+). Four different RFLP profile combinations (sets) were detected for all restriction endonucleases (HinfI, HhaI, and TaqI), of which three were characteristic of Anisakis typica, A. pegreffii, and A. physeteris, respectively. One profile set (for sample CA-2012) was linked to an ITS+ sequence ..