Abstract
Using genetic markers defined previously in the second internal transcribed spacer (ITS-2) of nuclear ribosomal DNA (rDNA), isotopic, and non-isotopic polymerase-chain-reaction-coupled single-strand conformation polymorphism (SSCP) were utilized to identify each of three anisakid species [Anisakis simplex (s.l.), Contracaecum osculatum (s.l.), and Hysterothylacium aduncum] from different host species and geographical locations in Poland and Sweden. While subtle microheterogeneity was observed within each of Anisakis simplex (s.l.) and H. aduncum, distinct SSCP profiles were displayed for each of the three species, allowing identification and differentiation of the three taxa. Subsequent sequencing of the ITS-1 and ITS-2 rDNA revealed that A. simplex (s.l.) represented Anisakis simplex s.s. and Contracaecum osculatum (s.l.) represented C. osculatum C. Application of the non-isotopic SSCP assay of ITS-2 to larval anisakid samples from different hosts and geographical locations in China revealed three distinct SSCP profiles, one of which was consistent with that of A. simplex (s.l.), and the other two had different SSCP profiles from that of C. osculatum C and H. aduncum. Sequencing of the ITS-1 and ITS-2 rDNA for representative Chinese anisakid samples examined revealed three anisakid species in China, i.e., Anisakis typica, Anisakis pegreffii, and Hysterothylacium sp. These molecular tools will be useful for identification and investigation of the ecology of anisakid nematodes in China and elsewhere.
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Acknowledgments
Project support was provided in part by a grant from the Natural Science Foundation of Guangdong Province (Team Project, grant No. 5200638) to XQZ, and from the Guangdong Entry-exit Inspection and Quarantine, China to HQY. The authors thank Dr. Bengt Sjtrand (Institute of Marine Research, Lysekil, Sweden) for providing samples of Anisakis simplex (s.l.) larvae from Skagerrak. The experiments comply with the current laws of the countries in which the experiments were performed.
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Zhu, X.Q., Podolska, M., Liu, J.S. et al. Identification of anisakid nematodes with zoonotic potential from Europe and China by single-strand conformation polymorphism analysis of nuclear ribosomal DNA. Parasitol Res 101, 1703–1707 (2007). https://doi.org/10.1007/s00436-007-0699-0
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DOI: https://doi.org/10.1007/s00436-007-0699-0