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Molecular approaches to differentiate three species of Nematodirus in sheep and goats from China based on internal transcribed spacer rDNA sequences

Published online by Cambridge University Press:  16 December 2013

G.H. Zhao ,
Y.Q. Jia ,
Q.Q. Bian ,
A.J. Nisbet ,
W.Y. Cheng ,
Y. Liu ,
Y.Q. Fang ,
X.T. Ma  and
S.K. Yu
G.H. Zhao*
Affiliation:
College of Veterinary Medicine, Northwest A&F University, Yangling712100, Shaanxi Province, PR China
Y.Q. Jia
Affiliation:
College of Veterinary Medicine, Northwest A&F University, Yangling712100, Shaanxi Province, PR China
Q.Q. Bian
Affiliation:
College of Veterinary Medicine, Northwest A&F University, Yangling712100, Shaanxi Province, PR China
A.J. Nisbet
Affiliation:
Parasitology Division, Moredun Research Institute, Pentlands Science Park, MidlothianEH26 0PZ, Scotland, UK
W.Y. Cheng
Affiliation:
College of Veterinary Medicine, Northwest A&F University, Yangling712100, Shaanxi Province, PR China
Y. Liu
Affiliation:
College of Veterinary Medicine, Northwest A&F University, Yangling712100, Shaanxi Province, PR China
Y.Q. Fang
Affiliation:
College of Veterinary Medicine, Northwest A&F University, Yangling712100, Shaanxi Province, PR China
X.T. Ma
Affiliation:
College of Veterinary Medicine, Northwest A&F University, Yangling712100, Shaanxi Province, PR China
S.K. Yu*
Affiliation:
College of Veterinary Medicine, Northwest A&F University, Yangling712100, Shaanxi Province, PR China
*
*Fax: +86-29-87081762, E-mail: [email protected] (G.H. Zhao); E-mail: [email protected] (S.K. Yu)
*Fax: +86-29-87081762, E-mail: [email protected] (G.H. Zhao); E-mail: [email protected] (S.K. Yu)
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Abstract

Internal transcribed spacer (ITS) rDNA sequences of three Nematodirus species from naturally infected goats or sheep in two endemic provinces of China were analysed to establish an effective molecular approach to differentiate Nematodirus species in small ruminants. The respective intra-specific genetic variations in ITS1 and ITS2 rDNA regions were 0.3–1.8% and 0–0.4% in N. spathiger, 0–6.5% and 0–5.4% in N. helvetianus, and 0–4.4% and 0–6.1% in N. oiratianus from China. The respective intra-specific variations of ITS1 and ITS2 were 1.8–4.4% and 1.6–6.1% between N. oiratianus isolates from China and Iran, 5.7–7.1% and 6.3–8.3% between N. helvetianus samples from China and America. For N. spathiger, compared with samples from China, sequence differences in ITS1 rDNA were 0.3–2.4% in isolates from America, 0.3–2.9% in New Zealand and 2.1–2.4% in Australia. Genetic variations in ITS2 rDNA of N. spathiger were 0–0.4% between samples from China and America, and 0–0.8% between samples from China and New Zealand. Using mutation sites, polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) and specific PCR techniques were developed to differentiate these three Nematodirus species. The specific PCR assay allowed the accurate identification of N. oiratianus from other common nematodes with a sensitivity of 0.69 pg and further examination of Nematodirus samples demonstrated the reliability of these two molecular methods.

Type
Research Papers
Information
Journal of Helminthology , Volume 89 , Issue 3 , May 2015 , pp. 259 - 266
Copyright
Copyright © Cambridge University Press 2013 

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Footnotes

†

G.H. Zhao and Y.Q. Jia contributed equally to this work.

References

Ahmed, M., Singh, M.N., Bera, A.K., Bandyopadhyay, S. & Bhattacharya, D. (2011) Molecular basis for identification of species/isolates of gastrointestinal nematode parasites. Asian Pacific Journal of Tropical Medicine 4, 589–593.Google Scholar
Audebert, F., Durette-Desset, M.C. & Chilton, N.B. (2000) Internal transcribed spacer rDNA can be used to infer the phylogenetic relationships of species within the genus Nematodirus (Nematoda: molineoidea). International Journal for Parasitology 30, 187–191.Google Scholar
Becklund, W.W. & Walker, M.L. (1967a) Nematodirus odocoilei sp. n. (Nematoda: Trichostrongylidae) from the black-tailed deer, Odocoileus hemionus, in North America. Journal of Parasitology 53, 392–394.Google Scholar
Becklund, W.W. & Walker, M.L. (1967b) Nematodirus of domestic sheep, Ovis aries, in the United States with a key to the species. Journal of Parasitology 53, 777–781.Google Scholar
Burland, T.G. (2000) DNASTAR's Lasergene sequence analysis software. Methods in Molecular Biology 132, 71–91.Google 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, 647–651.Google Scholar
Chitimia, L., Lin, R.Q., Cosoroaba, I., Braila, P., Song, H.Q. & Zhu, X.Q. (2009) Molecular characterization of hard ticks from Romania by sequences of the internal transcribed spacers of ribosomal DNA. Parasitology Research 105, 1479–1482.Google Scholar
Christensen, C.M., Zarlenga, D.S. & Gasbarre, L.C. (1994) Ostertagia, Haemonchus, Cooperia and Oesophagostomum: construction and characterization of genus-specific DNA probes to differentiate important parasites of cattle. Experimental Parasitology 78, 93–100.Google Scholar
Gasser, R.B. & Hoste, H. (1995) Genetic markers for closely-related nematodes. Molecular and Cell Probes 9, 315–320.Google Scholar
Gasser, R.B., Stevenson, L.A., Chilton, N.B., Nansen, P., Bucknell, D.G. & Beveridge, I. (1996) Species markers for equine strongyles detected in intergenic rDNA by PCR-RFLP. Molecular and Cellular Probes 10, 371–378.Google Scholar
Gasser, R.B., Rossi, L. & Zhu, X. (1999) Identification of Nematodirus species (Nematoda: Molineidae) from wild ruminants in Italy using ribosomal DNA markers. International Journal for Parasitology 29, 1809–1817.Google Scholar
Gasser, R.B., Bott, N.J., Chilton, N.B., Hunt, P. & Beveridge, I. (2008) Toward practical, DNA-based diagnostic methods for parasitic nematodes of livestock bionomic and biotechnological implications. Biotechnology Advances 26, 325–334.Google Scholar
Guindon, S. & Gascuel, O. (2003) A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Systems Biology 52, 696–704.Google Scholar
Justine, J.L., Briand, M.J. & Bray, R.A. (2012) A quick and simple method, usable in the field, for collecting parasites in suitable condition for both morphological and molecular studies. Parasitology Research 111, 341–351.Google Scholar
Lichtenfels, J.R. & Pilitt, P.A. (1983) Cuticular ridge patterns of Nematodirus (Nematoda: Trichostrongyloidea) parasitic in domestic ruminants of North America, with a key to species. Helminthology 50, 261–274.Google Scholar
Martínez-Ibeas, A.M., Martínez-Valladares, M., González-Lanza, C., Miñambres, B. & Manga-González, M.Y. (2011) Detection of Dicrocoelium dendriticum larval stages in mollusk and ant intermediate hosts by PCR, using mitochondrial and ribosomal internal transcribed spacer (ITS-2) sequences. Parasitology 24, 1–8.Google Scholar
Nadler, S.A., Hoberg, E.P., Hudspeth, D.S. & Rickard, L.G. (2000) Relationships of Nematodirus species and Nematodirus battus isolates (Nematoda: Trichostrongyloidea) based on nuclear ribosomal DNA sequences. Journal of Parasitology 86, 588–601.Google Scholar
Newton, L.A., Chilton, N.B., Beveridge, I., Hoste, H., Nansen, P. & Gasser, R.B. (1998a) Genetic markers for strongylid nematodes of livestock defined by PCR-based restriction analysis of spacer rDNA. Acta Tropica 69, 1–15.Google Scholar
Newton, L.A., Chilton, N.B., Beveridge, I. & Gasser, R.B. (1998b) Differences in the second internal transcribed spacer of four species of Nematodirus (Nematoda: Molineidae). International Journal for Parasitology 28, 337–341.Google Scholar
Nissen, S., Poulsen, I.H., Nejsum, P., Olsen, A., Roepstorff, A., Rubaire-Akiiki, C. & Thamsborg, S.M. (2011) Prevalence of gastrointestinal nematodes in growing pigs in Kabale District in Uganda. Tropical Animal Health and Production 43, 567–572.Google Scholar
Page, R.D. (1996) TreeView: an application to display phylogenetic trees on personal computers. Computer Applications in the Biosciences 12, 357–358.Google Scholar
Ross, J.L., Ivanova, E.S., Spiridonov, S.E., Waeyenberge, L., Moens, M., Nicol, G.W. & Wilson, M.J. (2010) Molecular phylogeny of slug-parasitic nematodes inferred from 18S rRNA gene sequences. Molecular Phylogenetics and Evolution 55, 738–743.CrossRefGoogle ScholarPubMed
Stringfellow, F. (1968) Bursal bosses as a diagnostic character in Nematodirus of domestic sheep, Ovis aries, in the United States. Journal of Parasitology 54, 891–895.Google Scholar
Tamura, K., Dudley, J., Nei, M. & Kumar, S. (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Molecular Biology and Evolution 24, 1596–1599.Google Scholar
Tariq, K.A., Chishti, M.Z., Ahmad, F. & Shawl, A.S. (2008) Epidemiology of gastrointestinal nematodes of sheep managed under traditional husbandry system in Kashmir valley. Veterinary Parasitology 158, 138–143.Google Scholar
Testini, G., Papini, R., Lia, R.P., Parisi, A., Dantas-Torres, F., Traversa, D. & Otranto, D. (2011) New insights into the morphology, molecular characterization and identification of Baylisascaris transfuga (Ascaridida, Ascarididae). Veterinary Parasitology 175, 97–102.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 25, 4876–4882.Google Scholar
Wang, C.R., Gao, J.F., Zhu, X.Q. & Zhao, Q. (2012) Characterization of Bunostomum trigonocephalum and Bunostomum phlebotomum from sheep and cattle by internal transcribed spacers of nuclear ribosomal DNA. Research in Veterinary Science 92, 99–102.Google Scholar
Wang, G.L., Liu, Z.Q., Nu, E. & Yi, M.T. (2010) The investigation of sheep disease in nine provinces of China. Grass-Feeding Livestock 146, 10–12 (in Chinese).Google Scholar
Wang, J.F. & Cao, R.F. (2008) The survey of gastrointestinal nematode infection in goats in Laiyang, Shangdong province. Shangdong Journal of Animal Husbandry and Veterinary Science 29, 42 (in Chinese).Google Scholar
Zhao, G.H., Hu, B., Song, J.K., Jia, Y.Q., Li, H.M., Wang, C.R., Lin, Q., Xu, Q.X., Yu, S.K. & Deng, Y. (2012a) Characterization of Oesophagostomum asperum and O. columbianum by internal transcribed spacers of nuclear ribosomal DNA. Journal of Helminthology 30, 1–8.Google Scholar
Zhao, G.H., Li, J., Song, H.Q., Li, X.Y., Chen, F., Lin, R.Q., Yuan, Z.G., Weng, Y.B., Hu, M., Zou, F.C. & Zhu, X.Q. (2012b) A specific PCR assay for the identification and differentiation of Schistosoma japonicum geographical isolates in mainland China based on analysis of mitochondrial genome sequences. Infection, Genetics and Evolution 12, 1027–1036.Google Scholar
Zheng, R.K., Zhao, S.H., Gao, H.Y., Geng, W.H. & Hu, M. (1997) The ecological study of larva of Nematodirus spp. in grazing sheep. Inner Mongolian Journal of Animal Sciences and Production 1, 9–14 (in Chinese).Google Scholar
Zhu, X.Q., Gasser, R.B., Podolska, M. & Chilton, N.B. (1998) Characterisation of anisakid nematodes with zoonotic potential by nuclear ribosomal DNA sequences. International Journal for Parasitology 28, 1911–1921.Google Scholar