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2003 Fiscal Year Final Research Report Summary

Basic research about the quick differential diagnosis of the Cryptosporidium and the prevention

Research Project

Project/Area Number 13660322
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied veterinary science
Research InstitutionOsaka Prefecture University

Principal Investigator

SASAKI Kazumi  Osaka Prefecture University, Graduate School of Agriculture and Life Sciences, Associate prof., 農学生命科学研究科, 助教授 (70211935)

Co-Investigator(Kenkyū-buntansha) MATSUBAYASHI Makoto  Osaka Prefecture University, Department of Food and Nutrition, Osaka Joshi-Gakuen Junior College, Assistant professor, 食物学科, 助手 (00321076)
TANI Hiroyuki  Osaka Prefecture University, Graduate School of agriculture and Life Sciences, Department of Veterinary Medicine, Assistant professor, 農学生命科学研究科, 助手 (00305658)
Project Period (FY) 2001 – 2003
KeywordsCryptosporidium / chicken monoclonal antibody / Diagnosis method / Giardia
Research Abstract

Applicants produced six sorts of chicken monoclonal antibodies (mAb) which used Eimeria acervulina as the antigen. Among these, mAb 6D-12-G10 showed the cross-reactivity with the sporozoites of Cryptosporidium parvum (following, CP) and C.muris. This time, cross-reactivities with the merozoites of CP cultivated in vitro were investigated using these chicken mAbs. Furthermore, it was examined whether the mAbs would inhibit host cell invasion of parasites. MAB 6D-12-G10 showed the cross-reactivities with merozoites of CP similarly, and the rate of invasion inhibition (%) was 75.4±4.0. It is suggested that anti-Eimeria chicken monoclonal antibody, 6D-12-Gb recognized the epitope related to host cell invasion of CP. Then, applicants tried to produce chicken monoclonal antibodies aiming at development of the simple differential-diagnosis method of Cryptosporidium (following, CR). In a field, CR shows cross-reactivities with the diatoms which inhabit in water resources using the commercial k … More its for CR detection. The immune tolerance to diatoms is made to gain by carrying out the immunization of the diatoms reported cross-reactivities with CR to chickens, then, the immunization of the oocysts of CR was carried out. The hybridoma was prepared by fusion of chicken myeloma cells with spleen cells isolated from the chicken gained the tolerance to diatoms and the immunity to CR, although production of a chicken monoclonal antibody was tried, a hybridoma stops an antibody production during the stage of single cloning and the production is under continuation.
Moreover, Giardia (following, Gi) trophozoites were detected in fecal samples from a pet dog with diarrhea. After treatment with metronidazole, this dog recovered from illness and Gi was not found in its fecal samples. When we examined the fecal samples of this dog in detail, CR oocysts were also found, and therefore this case was found to be a mixed infection with CR and Gi. In spite of the treatment with metronidazole, CR oocysts were still found in its fecal samples. This result suggests that metronidazole have no anti-cryptosporidial activity. CR isolate from this dog was identified as C.canis that was specific to dogs by PCR-RFLP analysis. This is the second case report about CR infection in pet dog and has documented the first finding of mixed infection with CR and Gi in dog in Japan. Less

  • Research Products

    (9 results)

All Other

All Publications (9 results)

  • [Publications] Matsubayashi M, et al.: "First description of Isospora spp. from the banded Mongoose, Mungos mungo, (Herpestidae) in Japan"Bulletin of Osaka Joshigakuen Junior College. 45. 43-45 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Constantinoiu C.C.et al.: "Analysis of cross-reactivity of five new chicken monoclonal antibodies which recognize the apical complex of Eimeria using confocal laser immunofluorescence assay."Veterinary Parasitology. 118(1-2). 29-35 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Matsubayashi M., et al.: "First record of Cryptosporidium infection in a racconn dog (Nyctereutes procyonoides viverrinus)."Veterinary Parasitology. 120(13). 171-175 (2004)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Constantinoiu C.C.et al.: "Characterization of stage-specific and cross-reactie antigens from Eimeria acervulina by chicken monoclonal antibodies."J.Vet.Med.Sci.. 66(4). 403-408 (2004)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Matsubayashi M., et al.: "飼い犬におけるGiardiaとCryptosporidiumの混合感染"日本獣医師会雑誌. (In press). (2004)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Matsubayashi M, et al.: "First description of Isospora spp.from the banded mongoose, Mungos mungo, (Herpestidae) in Japan"Bulletin of Osaka Joshigakuen Junior College. 45. 43-45 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Constantinoiu C.C., et al.: "Analysis of cross-reactivity of five new chicken monoclonal antibodies which recognize the apical complex of Eimeria using confocal laser immunofluorescence assay"Veterinary Parasitology. 118. 29-35 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Matsubayashi M., et al.: "First record of Cryptosporidium infection in a raccoon dog (Nyctereutes procyonoides viverrinus)"Veterinary Parasitology. 120. 171-175 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Constantinoiu C.C., et al.: "Characterization of stage-specific and cross-reactive antigens from Eimeria acervulina by chicken monoclonal antibodies"J.Vet.Med.Sci.. 66. 403-408 (2004)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2005-04-19  

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