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Identification and genetic characterization of A nisakis larvae from marine fishes in the S outh C hina S ea using an electrophoretic‐guided approach
Author(s) -
Zhang Luping,
Du Xiaojie,
An Ruiyong,
Li Liang,
Gasser Robin B.
Publication year - 2013
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201200493
Subject(s) - anisakis , biology , ribosomal dna , phylogenetic tree , clade , internal transcribed spacer , dna barcoding , genbank , anisakidae , zoology , genetics , evolutionary biology , larva , botany , gene , helminths
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 r RNA genes (= internal transcribed spacers, ITS +). Four different RFLP profile combinations (sets) were detected for all restriction endonucleases ( Hinf I, Hha I, and Taq I), 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 that was identical to a previously published sequence of Anisakis sp. (sample HC ‐2005; originating from the African shelf) and another sequence ( PH ‐2010; Madeira, Portugal). Phylogenetic analysis was carried out using the ITS + sequence data from this study and reference sequences from the GenBank database. Neighbor joining and maximum parsimony trees displayed three clades. Clades I and II included nine described species of Anisakis , including all type I and type II larvae; clade III represented some undescribed species of Anisakis . Morphological comparison showed that Anisakis sp. CA‐2012 was distinct from type I and type II larvae based on its tail shape and ratio of tail length to body length. The phylogenetic analysis and morphological characters suggest that Anisakis sp. CA‐2012 represents a new record, now called Anisakis type III larvae.