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Development of molecular diagnostic protocols for detecting three types of Entamoeba from diarrheal and asymptomatic pigs and environmental moist soils

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Abstract

Entamoeba suis and Entamoeba polecki subtypes (ST) 1 and 3 have recently been implicated in disease outbreaks in pigs. However, the distributions of these parasites in Japan and the potential sources of infection on farms still remain unclear. Here, we examined a farm of fattening/growing pigs with abnormal feces in Kagoshima Prefecture, Japan, and found the presence of parasites in the farm environment. Examination of intestinal tissues from pigs presenting with ulcerative colitis revealed a large number of trophozoites that had invaded the lesions. We identified single and mixed infections of E. suis and E. polecki ST1 and ST3 in paraffin sections or fecal samples from affected pigs. Two subtypes of Entamoeba were identified using four primer sets by PCR and sequencing. The parasites were detected in moist soil samples obtained around the drinking water source or puddles, implicating transmission of cysts via contaminated soils. Additionally, we found evidence of Entamoeba spp. and coinfections in surveyed pigs without any diarrhea at two neighboring farms. Our results establish methods for successfully identification of parasites, including cases in which multiple infections are present.

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Acknowledgements

We thank the staff of Kagoshima Central Livestock Hygiene Service Center for the collection of samples and Ms. R. Sekiguchi (Osaka Prefecture University) for assistance with molecular analyses. This work was partly supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science, and Technology (nos. 16 K15049 and 16H05803). We would also like to thank Editage (Tokyo) for English proofreading.

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Authors and Affiliations

  1. Kagoshima Central Livestock Hygiene Service Center, 1678 Yuda, Higashiichiki, Hioki, 899-2201, Japan

    Yoshimasa Hirashima, Takahisa Yano, Syoei Kitahara, Terunori Koreeda & Syunsuke Kamimura

  2. Veterinary Research Institute, Gannoruwa, Peradeniya, 20400, Sri Lanka

    Tilusha Manchanayake

  3. National Institute of Animal Health, National Agricultural and Food Research Organization, 3-1-5 Kannondai, Tsukuba, 305-0856, Japan

    Tilusha Manchanayake & Tomoyuki Shibahara

  4. Kanoya Meat Inspection Center, 3874-12, Kawanishi, Kanoya, 893-0032, Japan

    Takahisa Yano

  5. Livestock Industry Division of Kagoshima Prefecture, 10-1 Kamoike-shinmachi, Kagoshima, 890-8577, Japan

    Terunori Koreeda

  6. Department of Veterinary Science, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, 1-58 Rinku-oraikita, Izumisano, 598-8531, Japan

    Kazumi Sasai & Makoto Matsubayashi

Authors
  1. Yoshimasa Hirashima
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  2. Tilusha Manchanayake
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  3. Takahisa Yano
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  4. Syoei Kitahara
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  5. Terunori Koreeda
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  6. Syunsuke Kamimura
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  7. Kazumi Sasai
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  8. Makoto Matsubayashi
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  9. Tomoyuki Shibahara
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Corresponding authors

Correspondence to Makoto Matsubayashi or Tomoyuki Shibahara.

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Hirashima, Y., Manchanayake, T., Yano, T. et al. Development of molecular diagnostic protocols for detecting three types of Entamoeba from diarrheal and asymptomatic pigs and environmental moist soils. Parasitol Res 116, 2001–2007 (2017). https://doi.org/10.1007/s00436-017-5483-1

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  • DOI: https://doi.org/10.1007/s00436-017-5483-1

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