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Specific detection of Echinococcus spp. from the Tibetan fox (Vulpes ferrilata) and the red fox (V. vulpes) using copro-DNA PCR analysis

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Abstract

There are three Echinococcus species, Echinococcus granulosus, E. multilocularis, and E. shiquicus, which are distributed on the vast area of pastureland on the eastern Tibetan plateau in China. Tibetan foxes (Vulpes ferrilata) have been determined to be the main wild definitive host of E. multilocularis and E. shiquicus, but little information is available on the prevalence of these two parasites in Tibetan foxes. Consequently, the copro-prevalence of these parasites in foxes from the eastern Tibetan plateau was evaluated in this study. For each copro-DNA sample extracted from fox feces, a 133-bp segment of EgG1 Hae III was used to screen for infection with E. granulosus. Multiplex nested polymerase chain reaction (PCR) analysis was used to target an 874-bp segment of the mitochondrial COI gene to distinguish E. multilocularis and E. shiquicus. Among 184 fecal samples, 120 were from Tibetan foxes and six from red foxes (Vulpes vulpes). Of the fecal samples from Tibetan foxes, 74 (giving a copro-prevalence of 62 %) showed the presence of Echinococcus spp.: 23 (19 %) were found to contain E. multilocularis, 32 (27 %) E. shiquicus, and 19 (16 %) showed mixed infection with both E. multilocularis and E. shiquicus. Two fecal samples from red foxes were found to be infected with E. multilocularis. No fox feces were found to be infected with E. granulosus. Tests on zinc finger protein genes and a 105-bp fragment of the Sry gene found no significant difference in the prevalence of the two parasites between sexes. The efficiency of our multiplex nested PCR methods were compared with previous polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) methods and some problems associated with the copro-PCR were discussed.

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Acknowledgments

We thank Qiu Jiamin, Chen Xingwang, and other colleagues (Sichuan Center of Disease Control, Chengdu, China) for their help with field research. Colleagues from the Shiqu County Center of Disease Control provided us imperative logistical support during the field study. Dr. Wu Wei, Liu Xiaoqing, and Ma Bo (School of Life Science, East China Normal University, Shanghai, China) participated in the fieldwork. We are grateful to reviewers for their invaluable comments to improve this manuscript. This study was financially supported by grants from the National Science Foundation of China (NSFC no. 30870308 and 31071944), the Shanghai Rising-Star Program (STCSM no. 10QA1402200) and East China Normal University Innovative Program of Science Research for Ph.D. candidature (CX2011012).

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Authors and Affiliations

  1. School of Life Sciences, East China Normal University, No. 3663 North Zhongshan road, Shanghai, 200062, People’s Republic of China

    Weibin Jiang, Nan Liu, Gaotian Zhang & Zhenghuan Wang

  2. Center of Disease Control of Shiqu County, Ganzi, 627350, Sichuan Province, People’s Republic of China

    Pengcuo Renqing & Fei Xie

  3. Institute of Parasitic Diseases, Sichuan Centers for Disease Control and Prevention, No. 6 Middle School Road, Chengdu, 610041, Sichuan Province, People’s Republic of China

    Tiaoying Li

  4. Shanghai Key Laboratory of Urbanization and Ecological Restoration, East China Normal University, 3663 North Zhongshan road, Putuo, Shanghai, 200062, People’s Republic of China

    Zhenghuan Wang

  5. Shanghai Science and Technology Museum, 2000 Shijida Street, Pudong, Shanghai, 200127, People’s Republic of China

    Xiaoming Wang

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  1. Weibin Jiang
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Correspondence to Zhenghuan Wang or Xiaoming Wang.

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Jiang, W., Liu, N., Zhang, G. et al. Specific detection of Echinococcus spp. from the Tibetan fox (Vulpes ferrilata) and the red fox (V. vulpes) using copro-DNA PCR analysis. Parasitol Res 111, 1531–1539 (2012). https://doi.org/10.1007/s00436-012-2993-8

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