2003 Fiscal Year Final Research Report Summary
Basic research about the quick differential diagnosis of the Cryptosporidium and the prevention
| Project/Area Number |
13660322
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied veterinary science
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| Research Institution | Osaka Prefecture University |
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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)
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| 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)
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| Project Period (FY) |
2001 – 2003
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| Keywords | Cryptosporidium / chicken monoclonal antibody / Diagnosis method / Giardia |
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| 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
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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
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