Parasitology Research

, Volume 101, Issue 2, pp 351–356 | Cite as

Characterization of Oesophagostomum spp. from pigs in China by PCR-based approaches using genetic markers in the internal transcribed spacers of ribosomal DNA

  • Rui-Qing Lin
  • Xing-Quan ZhuEmail author
  • Dong-Xia Wei
  • Yan Deng
  • Wei Liu
  • Hui-Qun Song
  • An-Xing Li
  • Zhao-Rong Lun
Original Paper


In the present study, samples of Oesophagostomum spp. collected from pigs from different geographical localities in mainland China were characterized genetically by polymerase chain reaction-linked single-strand conformation polymorphism (PCR-SSCP) and restriction fragment length polymorphism (PCR-RFLP) techniques using genetic markers in the internal transcribed spacers (ITS) of nuclear ribosomal DNA (rDNA). The second internal transcribed spacer (ITS-2) was amplified from 51 individual nodule worms by PCR, and the amplicons were analyzed by SSCP. With the exception of slight microheterogeneity, SSCP analyses displayed two distinct banding profiles that allowed the identification of all Oesophagostomum spp. samples examined into two groups, the first one represented O. dentatum, and the second one may represent O. quadrispinulatum. Then, the entire ITS was amplified from individual samples, and the amplicons were digested with restriction endonuclease Pst I. The results of RFLP analyses were consistent with that of SSCP. Sequence analysis of ITS rDNA supported the identification and differentiation of Chinese Oesophagostomum spp. samples into two species, namely, O. dentatum and O. quadrispinulatum. These PCR-based approaches provide useful complementary tools to traditional methods for the accurate identification of Oesophagostomum spp. (irrespective of developmental stage) and have implications for studying the ecology and population genetic structures of these parasites and for the prevention and control of the diseases they cause.


Polymerase Chain Reaction Product Internal Transcribe Spacer Restriction Fragment Length Polymorphism Analysis Dongting Lake SSCP Analysis 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



Project support was provided to XQZ in part by grants from the China National Science Funds for Distinguished Young Scientists (grant no.30225033) and from the Excellent Young Teachers’ Program of Ministry of Education, China (no. [2002] 350).

The experiments comply with the current laws of the country in which the experiments were performed.


  1. Andree KB, E1-Matbouli M, Hoffman RW, Hedrick RP (1999) Comparison of 18S and ITS-1 rDNA sequences of selected geographic isolates of Myxobolus cerebralis. Int J Parasitol 29:771–775PubMedCrossRefGoogle Scholar
  2. Boes J, Willingham AL III, Shi FH, Hu XG, Eriksen L, Nansen P, Stewart TB (2000) Prevalence and distribution of pig helminthes in the Dongting Lake Region (Hunan Province) of the People’s Republic of China. J Helminthol 74:45–52PubMedGoogle Scholar
  3. Chilton NB, Gasser RB, Beveridge I (1995) Differences in a ribosomal DNA sequence of morphologically indistinguishable species within the Hypodontus macropi complex (Nematoda: Strongyloidea). Int J Parasitol 25:647–651PubMedCrossRefGoogle Scholar
  4. Colenman AW (2005) Paramecium Aurelia revisited. J Eukaryot Microbiol 52:68–77CrossRefGoogle Scholar
  5. Cutillas C, Guevara-Martínez D, Oliveros R, Arias P, Guevara DC (1999) Characterization of porcine and ovine Oesophagostomum spp. by isoenzymatic patterns and restriction-fragment-length polymorphisms (RFLPs). Acta Trop 73:59–71PubMedCrossRefGoogle Scholar
  6. Elbadri GAA, De Ley P, Waeyenberge L, Vierstraete A, Moens M, Vanfleteren J (2002) Intraspecific variation in Radopholus similes isolates assessed with restriction fragment length polymorphism and DNA sequencing of the internal transcribed spacer region of the ribosomal RNA cistron. Int J Parasitol 32:199–220PubMedCrossRefGoogle Scholar
  7. Gasser RB, Woods WG, Bjørn H (1998) Distinguishing Oesophagostomum dentatum from Oesophagostomum quadrispinulatum developmental stages by a single-strand conformation polymorphism method. Int J Parasitol 28:1903–1909PubMedCrossRefGoogle Scholar
  8. Hong M, Wang CJ, Zhang YZ, Xia BY, Wang WD (2000) Investigation on the prevalence of parasites in pigs in Tongren Administrative Region, Guizhou Province, People’s Republic of China. Chin J Vet Parasitol 8(2):28, (in Chinese)Google Scholar
  9. Huang WY, He B, Wang CR, Zhu XQ (2004) Characterisation of Fasciola species from Mainland China by ITS-2 ribosomal DNA sequence. Vet Parasitol 120:75–83PubMedCrossRefGoogle Scholar
  10. Jing A, Li M, Wang LX, Su JH, Nia K, Yu BH, Pan HM, He L (2002) Investigation on the prevalence of parasites in pigs in the City of Jiangjing, Sichuan Province, People’s Republic of China. Chin J Vet Parasitol 10(2):24–25, (in Chinese)Google Scholar
  11. Joachim A, Dülmer N, Daugschies A, Roepstorff A (2001) Occurrence of helminthes in pig fattening units with different management systems in Northern Germany. Vet Parasitol 96:135–146PubMedCrossRefGoogle Scholar
  12. Legesse M, Erko B (2004) Zoonotic intestinal parasites in Papio anubis (baboon) and Cercopithecus aethiops (vervet) from four localities in Ethiopia. Acta Trop 90:231–236PubMedCrossRefGoogle Scholar
  13. Li AX, Amello SD, Paggl L, He F, Gasser RB, Lun ZR, Abollo E, Turchetto M, Zhu XQ (2005a) Genetic evidence for the existence of sibling species within Contracaecum rudolphii Hartwich, 1964 and the validity of Contracaecum septentrionale Kreis, 1955 (Nematoda: Anisakidae). Parasitol Res 96:361–366PubMedCrossRefGoogle Scholar
  14. Li ZM, Xiao X, Yan DW, Li JS (2005b) Investigation of parasites of pigs in Yunnan Province. Acta Ecol Anim Domast 26(6):67–70, (in Chinese)Google Scholar
  15. Li MW, Zhu XQ, Gasser RB, Lin RQ, Sani RA, Lun ZR, Jacobs DE (2006) The occurrence of Toxocara malaysiensis in cats in China, confirmed by sequence-based analyses of ribosomal DNA. Parasitol Res 99:554–557PubMedCrossRefGoogle Scholar
  16. Liu CQ, Lu J (2002) Investigation on the prevalence of parasites in intensively raised pigs in Yulin Administrative Region, Guangxi Zhuang Nationality Autonomous Region, People’s Republic of China. Guangxi J Anim Sci Vet Med 18(6):11–12, (in Chinese)Google Scholar
  17. Lu JJ, Jin JS, Zhang L, Yao XY, Jin ZQ (2002) Keys to and illustrative descriptions for parasitic nematodes of humans and animals. Chinese Press of Agricultural Science and Technology, Beijing, China, (in Chinese)Google Scholar
  18. Meinilä M, Kuusela J, Ziętara MS, Lumme J (2004) Initial steps of speciation by geographic isolation and host switch in salmonid pathogen Gyrodactylus salaries (Monogenea: Gyrodactylidae). Int J Parasitol 34:515–526PubMedCrossRefGoogle Scholar
  19. Newton LA, Chilton NB, Monti JR, Bjørn H, Várady M, Christensen CM, Gasser RB (1997) Rapid PCR-based delineation of the porcine nodular worms, Oesophagostomum dentatum and O. quadrispinulatum. Mol Cell Probes 11:149–153PubMedCrossRefGoogle Scholar
  20. Permin A, Yelifari L, Bloch P, Steenhard N, Hansen NP, Nansen P (1999) Parasites in cross-bred pigs in the Upper East Region of Ghana. Vet Parasitol 87:63–71PubMedCrossRefGoogle Scholar
  21. Poelvoorde J (1978) Some characteristics of a laboratory produced strain of Oesophagostomum dentatum. Vet Parasitol 4:369–376CrossRefGoogle Scholar
  22. Polderman AM, Blotkamp J (1995) Oesophagostomum infections in humans. Parasitol Today 11:451–456PubMedCrossRefGoogle Scholar
  23. Polderman AM, Anemana SD, Asigri V (1999) Human oesophagostomiasis: a regional public health problem in Africa. Parasitol Today 15:129–130PubMedCrossRefGoogle Scholar
  24. Roepstorff A, Nilsson O, Oksanen A, Gjerde B, Richter SH, Örtenberg E, Christensson D, Martinsson KB, Bartlett PC, Nansen P, Eriksen L, Helle O, Nikander S, Larsen K (1998) Intestinal parasites in swine in the Nordic countries: prevalence and geographical distribution. Vet Parasitol 76:305–319PubMedCrossRefGoogle Scholar
  25. Skrjabin KI, Shikhobalova NP, Mozgovoi AA (1991) Suborder. Ascaridata Skrjabin, 1915. In: Skrjabin KI (ed) Key to parasitic nematodes. Amerind, New DelhiGoogle Scholar
  26. Stewart TB, Gasbarre LC (1989) The veterinary importance of nodular worms (Oesophagostomum spp). Parasitol Today 5:209–213PubMedCrossRefGoogle Scholar
  27. Storey PA, Faile G, Hewitt E, Yelifari L, Polderman AM, Magnussen P (2000) Clinical epidemiology and classification of human oesophagostomiasis. Trans R Soc Trop Med Hyg 94:177–182PubMedCrossRefGoogle Scholar
  28. Wan ZX, Wang Q, Zeng ZM, Hu X, He H, Li Z, Yang YJ, Zhang Q, Li CJ, Hu YZ, Wu JH, Long HY, Yang LQ, Hu ZM, Zou MH (2002) Investigation on the prevalence of parasites in pigs in Bijie Administrative Region, Guizhou Province, People’s Republic of China. Chin J Vet Parasitol 10(3):16–18, (in Chinese)Google Scholar
  29. Weng YB, Hu YJ, Li Y, Li BS, Lin RQ, Xie DH, Gasser RB, Zhu XQ (2005) Survey of intestinal parasites from intensive pig farms in Guangdong Province, People’s Republic of China. Vet Parasitol 127:333–336PubMedCrossRefGoogle Scholar
  30. Wu XY, Chilton NB, Zhu XQ, Xie MQ, Li AX (2005) Molecular and morphological evidence indicates that Pseudorhabdosynochus lantauensis (Monogenea: Diplectanidae) represents two species. Parasitology 130:669–677PubMedCrossRefGoogle Scholar
  31. Zhou WZ, Zhou JF, Bi CA, Liang ZS, Guo CR, Song WX, Lv CY (2001) Investigation on the prevalence of intestinal parasites in pigs in the City of Yuxi, Yunnan Province, People’s Republic of China. Chin J Vet Parasitol 9(3):24–25, (in Chinese)Google Scholar
  32. Zhu XQ, D’Amelio S, Paggi L, Gasser RB (2000) Assessing sequence variation in the internal transcribed spacers of ribosomal DNA within and among members of the Contracaecum osculatum complex (Nematoda: Ascaridoidea: Anisakidae). Parasitol Res 86:677–683PubMedCrossRefGoogle Scholar
  33. Zhu XQ, D’Amelio S, Hu M, Paggi L, Gasser RB (2001) Electrophoretic detection of population variation within Contracaecum ogmorhini (Nematoda: Ascaridoidea: Anisakidae). Electrophoresis 22:1930–1934PubMedCrossRefGoogle Scholar
  34. Zhu XQ, D’Amelio S, Palm HW, Paggi L, George-Nascimento M, Gasser RB (2002) SSCP-based identification of members within the Pseudoterranova decipiens complex (Nematoda: Ascaridoidea: Anisakidae) using genetic markers in the internal transcribed spacers of ribosomal DNA. Parasitology 124:615–623PubMedCrossRefGoogle Scholar
  35. Zhu XQ, D’Amelio S, Gasser RB, Yang TB, Paggi L, He F, Lin RQ, Song HQ, Lin A, Li AX (2007) Practical PCR tools for the delineation of Contracaecum rudolphii A and Contracaecum rudolphii B (Ascaridoidea: Anisakidae) using genetic markers in nuclear ribosomal DNA. Mol Cell Probes 21:97–102PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Rui-Qing Lin
    • 1
  • Xing-Quan Zhu
    • 1
    Email author
  • Dong-Xia Wei
    • 2
  • Yan Deng
    • 3
  • Wei Liu
    • 4
  • Hui-Qun Song
    • 1
  • An-Xing Li
    • 5
  • Zhao-Rong Lun
    • 5
  1. 1.College of Veterinary MedicineSouth China Agricultural UniversityGuangzhouPeople’s Republic of China
  2. 2.Department of Veterinary ScienceJiangsu Animal Husbandry and Veterinary CollegeTaizhouPeople’s Republic of China
  3. 3.General Laboratory of Inspection and QuarantineGuangzhou Airport Entry–Exit Inspection and Quarantine BureauGuangzhouPeople’s Republic of China
  4. 4.College of Animal Science and TechnologyHunan Agricultural UniversityChangshaPeople’s Republic of China
  5. 5.School of Life SciencesZhongshan (Sun Yat-Sen) UniversityGuangzhouPeople’s Republic of China

Personalised recommendations