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Genetic variation among Clonorchis sinensis isolates from different geographic regions in China revealed by sequence analyses of four mitochondrial genes

Published online by Cambridge University Press:  12 December 2011

G.H. Liu
Affiliation:
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province730046, PR China College of Veterinary Medicine, Hunan Agricultural University, Changsha, Hunan Province410128, PR China
B. Li
Affiliation:
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province730046, PR China Guangdong HuanongWens Animal Husbandry Co. Ltd, Yunfu, Guangdong Province527400, PR China College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province510642, PR China
J.Y. Li
Affiliation:
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province730046, PR China College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province510642, PR China
H.Q. Song
Affiliation:
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province730046, PR China
R.Q. Lin
Affiliation:
College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province510642, PR China
X.Q. Cai
Affiliation:
Zhongshan Entry-Exit Inspection and Quarantine Bureau, Zhongshan, Guangdong Province528403, PR China
F.C. Zou
Affiliation:
College of Animal Science and Technology, Yunnan Agricultural University, Kunming, Yunnan Province650201, PR China
H.K. Yan
Affiliation:
College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province510642, PR China
Z.G. Yuan
Affiliation:
College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province510642, PR China
D.H. Zhou
Affiliation:
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province730046, PR China
X.Q. Zhu*
Affiliation:
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province730046, PR China College of Veterinary Medicine, Hunan Agricultural University, Changsha, Hunan Province410128, PR China College of Animal Science and Technology, Yunnan Agricultural University, Kunming, Yunnan Province650201, PR China
*
*Fax: +86 (931) 8340977 E-mail: [email protected]

Abstract

The present study examined sequence variation in four mitochondrial (mt) genes, namely cytochrome c oxidase subunits 1 (cox1) and 2 (cox2), and NADH dehydrogenase subunits 1 and 2 (nad1 and nad2) among Clonorchis sinensis isolates from different endemic regions in China, and their phylogenetic relationships with other zoonotic trematodes were reconstructed. A portion of the cox1 and cox2 genes (pcox1 and pcox2), and nad1 and nad2 genes (pnad1 and pnad2) were amplified separately from individual liver flukes by polymerase chain reaction (PCR) and the amplicons were subjected to sequencing from both directions. The intra-specific sequence variations within C. sinensis were 0–1.6% for pcox1, 0–1.4% for pcox2, 0–0.9% for pnad1 and 0–1.0% for pnad2. Phylogenetic analyses based on the combined sequences of pcox1, pcox2, pnad1 and pnad2 revealed that all the C. sinensis isolates grouped together and were closely related to Opisthorchis felineus. These findings revealed the existence of intra-specific variation in mitochondrial DNA (mtDNA) sequences among C. sinensis isolates from different geographic regions, and demonstrated that mtDNA sequences provide reliable genetic markers for phylogenetic studies of zoonotic trematodes.

Type
Research Papers
Copyright
Copyright © Cambridge University Press 2011

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References

Ai, L., Chen, M.X., Alasaad, S., Elsheikha, H.M., Li, J., Li, H.L., Lin, R.Q., Zou, F.C., Zhu, X.Q. & Chen, J.X. (2011) Genetic characterization, species differentiation and detection of Fasciola spp. by molecular approaches. Parasites and Vectors 4, 101.CrossRefGoogle ScholarPubMed
Cai, X.Q., Liu, G.H., Song, H.Q., Wu, C.Y., Zou, F.C., Yan, H.K., Yuan, Z.G., Lin, R.Q. & Zhu, X.Q. (2012) Sequences and gene organization of the mitochondrial genomes of the liver flukes Opisthorchis viverrini and Clonorchis sinensis (Trematoda). Parasitology Research 31 May (Epub ahead of print).CrossRefGoogle ScholarPubMed
Cerutti, M.C., Citterio, C.V., Bazzocchi, C., Epis, S., D'Amelio, S., Ferrari, N. & Lanfranchi, P. (2010) Genetic variability of Haemonchus contortus (Nematoda: Trichostrongyloidea) in alpine ruminant host species. Journal of Helminthology 84, 276283.CrossRefGoogle ScholarPubMed
Chilton, N.B., Gasser, R.B. & Beveridge, I. (1995) Differences in a ribosomal DNA sequence of morphologically indistinguishable species within the Hypodontus macropi complex (Nematoda: Strongyloidea). International Journal for Parasitology 25, 647651.CrossRefGoogle Scholar
Dai, R.S., Liu, G.H., Song, H.Q., Lin, R.Q., Yuan, Z.G., Li, M.W., Huang, S.Y., Liu, W. & Zhu, X.Q. (2012) Sequence variability in two mitochondrial DNA regions and internal transcribed spacer among three cestodes infecting animals and humans from China. Journal of Helminthology 86, 245251.CrossRefGoogle ScholarPubMed
Gasser, R.B. & Newton, S.E. (2000) Genomic and genetic research on bursate nematodes: significance, implications and prospects. International Journal for Parasitology 30, 509534.CrossRefGoogle ScholarPubMed
Guindon, S. & Gascuel, O. (2003) A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Systematic Biology 52, 696704.CrossRefGoogle ScholarPubMed
Kaewkes, S. (2003) Taxonomy and biology of liver flukes. Acta Tropica 88, 177186.CrossRefGoogle ScholarPubMed
Kang, S., Sultana, T., Loktev, V.B., Wongratanacheewin, S., Sohn, W.M., Eom, K.S. & Park, J.K. (2008) Molecular identification and phylogenetic analysis of nuclear rDNA sequences among three opisthorchid liver fluke species (Opisthorchiidae: Trematoda). Parasitology International 57, 191197.CrossRefGoogle ScholarPubMed
Katokhin, A.V., Shekhovtsov, S.V., Konkow, S., Yurlova, N.I., Serbina, E.A., Vodianitskai, S.N., Fedorov, K.P., Loktev, V.B., Muratov, I.V., Ohyama, F., Makhnev, T.V., Pel'tek, S.E. & Mordvinov, V.A. (2008) Assessment of the genetic distinctions of Opisthorchis felineus from O. viverrini and Clonorchis sinensis by ITS2 and CO1 sequences. Doklady Biochemistry and Biophysics 421, 214217.CrossRefGoogle ScholarPubMed
Lai, D.H., Wang, Q.P., Chen, W., Cai, L.S., Wu, Z.D., Zhu, X.Q. & Lun, Z.R. (2008) Molecular genetic profiles among individual Clonorchis sinensis adults collected from cats in two geographic regions of China revealed by RAPD and MGE-PCR methods. Acta Tropical 107, 213216.CrossRefGoogle ScholarPubMed
Li, J., Zhao, G.H., Zou, F.C., Mo, X.H., Yuan, Z.G., Ai, L., Li, H.L., Weng, Y.B., Lin, R.Q. & Zhu, X.Q. (2010) Combined mitochondrial 16S and 12S rDNA sequences: an effective genetic marker for inter-species phylogenetic analysis of zoonotic trematodes. Parasitology Research 107, 561569.CrossRefGoogle ScholarPubMed
Li, M.W., Lin, R.Q., Song, H.Q., Sani, R.A., Wu, X.Y. & Zhu, X.Q. (2008) Electrophoretic analysis of sequence variability in three mitochondrial DNA regions for ascaridoid parasites of human and animal health significance. Electrophoresis 29, 29122917.CrossRefGoogle ScholarPubMed
Lin, R.Q., Qiu, L.L., Liu, G.H., Wu, X.Y., Weng, Y.B., Xie, W.Q., Hou, J., Pan, H., Yuan, Z.G., Zou, F.C., Hu, M. & Zhu, X.Q. (2011) Characterization of the complete mitochondrial genomes of five Eimeria species from domestic chickens. Gene 480, 2833.CrossRefGoogle ScholarPubMed
Liu, G.H., Lin, R.Q., Li, M.W., Liu, W., Liu, Y., Yuan, Z.G., Song, H.Q., Zhao, G.H., Zhang, K.X. & Zhu, X.Q. (2011) The complete mitochondrial genomes of three cestode species of Taenia infecting animals and humans. Molecular Biology Reports 38, 22492256.CrossRefGoogle ScholarPubMed
Lun, Z.R., Gasser, R.B., Lai, D.H., Li, A.X., Zhu, X.Q., Yu, X.B. & Fang, Y.Y. (2005) Clonorchiasis: a key foodborne zoonosis in China. Lancet Infectious Diseases 5, 3141.CrossRefGoogle ScholarPubMed
Page, R.D. (1996) TREEVIEW: an application to display phylogenetic trees on personal computers. Computer Applications in the Biosciences 12, 357358.Google ScholarPubMed
Park, G.M. (2007) Genetic comparison of liver flukes, Clonorchis sinensis and Opisthorchis viverrini, based on rDNA and mtDNA gene sequences. Parasitology Research 100, 351357.CrossRefGoogle ScholarPubMed
Posada, D. (2008) JModelTest phylogenetic model averaging. Molecular Biology and Evolution 25, 12531256.CrossRefGoogle ScholarPubMed
Ronquist, F. & Huelsenbeck, J.P. (2003) MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19, 15721574.CrossRefGoogle ScholarPubMed
Saijuntha, W., Sithithaworn, P., Wongkham, S., Laha, T., Chilton, N.B., Petney, T.N., Barton, M. & Andrews, R.H. (2008) Mitochondrial DNA sequence variation among geographical isolates of Opisthorchis viverrini in Thailand and Lao PDR, and phylogenetic relationships with other trematodes. Parasitology 135, 14791486.CrossRefGoogle ScholarPubMed
Shekhovtsov, S.V., Katokhin, A.V., Kolchanov, N.A. & Mordvinov, V.A. (2010) The complete mitochondrial genomes of the liver flukes Opisthorchis felineus and Clonorchis sinensis (Trematoda). Parasitology International 59, 100103.CrossRefGoogle ScholarPubMed
Shin, H.R., Oh, J.K., Masuyer, E., Curado, M.P., Bouvard, V., Fang, Y.Y., Wiangnon, S., Sripa, B. & Hong, S.T. (2010) Epidemiology of cholangiocarcinoma: an update focusing on risk factors. Cancer Sciences 101, 579585.CrossRefGoogle ScholarPubMed
Swofford, D.L. (2002) PAUP*: Phylogenetic analysis using parsimony (and other methods). Sunderland, Massachusetts, Sinauer Associates.Google Scholar
Thompson, J.D., Gibson, T.J., Plewniak, F., Jeanmougin, F. & Higgins, D.G. (1997) The Clustal X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research 24, 48764882.CrossRefGoogle Scholar
Zhao, G.H., Mo, X.H., Zou, F.C., Weng, Y.B., Lin, R.Q., Xia, C.M. & Zhu, X.Q. (2009) Genetic variability among Schistosoma japonicum isolates from different endemic regions in China revealed by sequences of three mitochondrial DNA genes. Veterinary Parasitology 162, 6774.CrossRefGoogle Scholar