Development of discrimination methods among bovine Eimeria species and its utilization to diagnose bovine coccidiosis

• Project/Area Number: 17K08073
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Veterinary medical science
• Research Institution: Iwate University
• Principal Investigator: Itagaki Tadashi 岩手大学, 農学部, 教授 (80203074)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 牛コクシジウム症 / nested multiplex PCR / Eimeria / 種識別 / 牛 / コクシジウム症 / ITS2 / nested multiplex PCR法 / 種識別診断法 / コクシジウム / ウシ / 遺伝子診断

Outline of Final Research Achievements

The purpose of this study was to develop technique to discriminate between bovine Eimeria spp. and diagnostic method for bovine coccidiosis. Ten Eimeria species, E. cylidrica、E. subspherica、E. wyomingensis、E. auburnensis、E. ellipsoidaris、E. zuernii、E. alabamensis、E. bovis、E. bukidnensis、E. canadensis were discriminated by newly developed nested multiplex PCR available for field samples .

Academic Significance and Societal Importance of the Research Achievements

牛のコクシジウム症はEimeria属原虫の感染に起因し、世界の畜産業に甚大な経済損失を与えている.その診断には糞便中オーシストの検出とその形態的種同定が求められる。しかし、検査には寄生虫学的熟練と時間を要し、新たな検査法の開発が望まれている。本研究では、牛寄生Eimeria10種を遺伝子領域のDNA塩基配列の違いで識別できることを明らかにし、 複数種の同時検出も可能なnested multiplex PCR法の開発に成功した。これにより、特別知識を要しないPCR検査法で牛のコクシジウム症を診断することが可能になった。

Research Products

• [Journal Article] Phylogenetic relationships between Lymnaeidae in relation to infection with Fasciola sp. in Hokkaido, Japan. (2020)
  • Author(s): Yuma Ohari, Kei Hayashi, Uday Kumar Mohanta, Tatsuo Oshida, and Tadashi Itagaki
  • Journal Title: Mollus. Res. Volume: 40 Pages: 160-168
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] The prevalence and molecular characterization of Acarapis woodi and Varroa destructor mites in honeybees in the Tohoku region of Japan. (2020)
  • Author(s): Shunsuke Takashima, Yuma Ohari, Tadashi Itagaki
  • Journal Title: Parasitol Int Volume: 75 Pages: 102052-102052
  • Peer Reviewed
• [Journal Article] Multigene typing and phylogenetic analysis of Fasciola from endemic foci in Iran. (2020)
  • Author(s): Ehsan Javanmard, Yuma Ohari, Amir Sadeghi, Kourosh Cheraghipour,Hamid Asadzadeh Aghdaei , Hamed Mirjalali , Mohammad Reza Zali , Tadashi Itagaki
  • Journal Title: Inf. Gen. Evol. Volume: 80 Pages: 104202-104202
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] First report of Paragonimus skrjabini miyazakii metacercariae in Geothelphusa dehaani (Sawagani) occurring in Iwate Prefecture, Japan. (2019)
  • Author(s): Yuma Ohari, Yuma Suzuki, Toshiyuki Shibahara and Tadashi Itagaki
  • Journal Title: J Vet Med Sci Volume: 81(8) Pages: 1109-1112
  • Peer Reviewed
• [Journal Article] Morphological and genetic characterization of green-banded broodsacs of T Leucochloridium (Leucochloridiidae: Trematoda) sporocysts detected in Succinea lauta in Hokkaido, Japan (2019)
  • Author(s): Yuma Ohari, Yasuhiro Kuwahara, Tadashi Itagaki
  • Journal Title: Parasitology International Volume: 68 Pages: 53-56
  • Peer Reviewed
• [Journal Article] Characterization of Echinostoma revolutum and Echinostoma robustum from ducks in Bangladesh based on morphology, nuclear ribosomal ITS2 and mitochondrial nad1 sequences (2019)
  • Author(s): Uday Kumar Mohanta, Takuya Watanabe, Anisuzzaman, Yuma Ohari, Tadashi Itagaki
  • Journal Title: Parasitology International Volume: 69 Pages: 1-7
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Hybrid origin of Asian aspermic Fasciola flukes is confirmed by analyzing two single-copy genes, pepck and pold. (2018)
  • Author(s): Hayashi K, Ichikawa-Seki M, Mohanta UK, Shoriki T, Chaichanasak P, Itagaki T.
  • Journal Title: Journal of Veterinary Medical Science Volume: 80 Pages: 98-102
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Phylogenetic relationships between Dicrocoelium chinensis populations in Japan and China based on mitochondrial nad1 gene sequences. (2017)
  • Author(s): Hayashi K, Tang W, Ohari Y, Ohtori M, Mohanta UK, Matsuo K, Sato H, Itagaki T
  • Journal Title: Parasitol Res. Volume: 116 Issue: 9 Pages: 2605-2609
  • DOI: 10.1007/s00436-017-5557-0
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] First report of <i>Fasciola</i> larva infection in <i>Galba truncatula</i> (Müller, 1774) (Gastropoda, Lymnaeidae) occurring in the natural environment in Hokkaido, Japan (2017)
  • Author(s): Ohari Y, Hayashi K, Mohanta UK, Kuwahara Y, Itagaki T.
  • Journal Title: Journal of Veterinary Medical Science Volume: 79 Issue: 8 Pages: 1381-1383
  • DOI: 10.1292/jvms.17-0215
  • NAID: 130005950122
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Molecular characterization and phylogeny of <i>Linguatula serrata</i> (Pentastomida: Linguatulidae) based on the nuclear 18S rDNA and mitochondrial cytochrome <i>c</i> oxidase I gene (2017)
  • Author(s): Mohanta UK, Itagaki T.
  • Journal Title: Journal of Veterinary Medical Science Volume: 79 Issue: 2 Pages: 398-402
  • DOI: 10.1292/jvms.16-0508
  • NAID: 130005397098
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Molecular and phylogenetic analyses of the liver amphistome Explanatum explanatum (Creplin, 1847) Fukui, 1929 in ruminants from Bangladesh and Nepal based on nuclear ribosomal ITS2 and mitochondrial nad1 sequences. (2017)
  • Author(s): Mohanta UK, Rana HB, Devkota B, Itagaki T.
  • Journal Title: J Helminthol Volume: 91 Issue: 4 Pages: 497-503
  • DOI: 10.1017/s0022149x16000420
  • Peer Reviewed / Int'l Joint Research