In order to identify the species of nematodes isolated from dolphin fecal samples,the morphology of the isolated nematodes is first identified.The parasite's body length was observed to be 120-140 mm long,with a cylindrical shape and slender ends.Its appearance was milky white and translucent,and its internal structure was observed to be yellowish under the micro-scope.It can be seen that the front end of the esophagus is thin and gradually expands back-wards.We extracted the genomic DNA of the isolated worm and amplified the 28S rRNA D2-D3 gene and ITS gene fragments by PCR for molecular identification.The PCR results showed that the amplification products of the 28S rRNA D2-D3 gene and ITS gene were 797 and 840 bp,re-spectively.BLAST alignment results showed that the sequence similarity between the 28S rRNA D2-D3 gene(GenBank accession number OR345165)and the Anisakis typical(GenBank acces-sion number HF911524.1)was 95.36％.The sequence similarity between the ITS gene(Gen-Bank accession number OR345164)and the Anisakis typical(GenBank accession number MF668919.1)was 97.11％,with a 91.90％sequence identity between the ITS-1 region and the Anisakis typical(GenBank accession number MF668919.1).There is a 94.97％sequence identi-ty between the ITS-2 region and the Anisakis typical(GenBank accession number JN968938.1)and a 100.00％sequence identity between the 5.8S rRNA region and the Anisakis typical(Gen-Bank accession number OQ244221.1).Phylogenetic analyses based on the ITS gene and the 28S rRNA D2-D3 gene showed that the parasite samples isolated in this study were genetically closest to the Anisakis typical,and on the same branch.In summary,the isolated parasite samples ob-tained from this study should be the larvae of Anisakis typical based on microscopic examination and molecular analysis.
Anisakiszoonosismorphological observationmolecular identificationphylogenet-ic tree
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