| Project/Area Number |
06660397
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Applied veterinary science
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| Research Institution | Obihiro University Agriculture and Veterinary Medicine |
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Principal Investigator |
IGARASHI Ikuo Obihiro Univ., Res.Ctr.Protozoan Mol.Immunol., Assoc.Prof., 原虫病分子免疫研究センター, 助教授 (80159582)
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| Co-Investigator(Kenkyū-buntansha) |
OMATA Yoshitaka Obihiro Univ., Vet.Physiol., Assoc.Prof., 畜産学部, 助教授 (10132987)
SAITO Atsushi Obihiro Univ., Vet.Physiol., Prof., 畜産学部, 教授 (10002263)
SUZUKI Naoyoshi Obihiro Univ., Res.Ctr.Protozoan Mol.Immunol., Emeritus.Prof., 名誉教授 (10003071)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1995: ¥500,000 (Direct Cost: ¥500,000)
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| Keywords | Babesia ovata / Babesia equi / Babesia caballi / In vitro culture / IFAT / Antibody survy / Monoclonal antibody / 酵素抗体法(ELISA) / ウマバベシア / in vitro連続培養 / 低酸素 / 凍結保存 |
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| Research Abstract |
Babesiosis in cattle and equine is endemic and causes enormous economic losses throughout most of the tropical and subtropical regions of the world. In this study, establishment of continuous cultivation of bovine and equine Babesia parasites were examined.Babesia ovata was initiated their multiplication within adult bovine RBC in Medium 199 supplemented with 40% adult bovine serum under a low oxygen atmosphere, 5% O2,5% CO2 and 90% N2.Babesia equi and B.caballi were continuously cultured in vitro in M199 and RPMI 1640 medium respectively, supplemented with 2 mM L-glutamine, 20 mM HEPES and 40% adult horse serum in a low oxygen atmosphere (2%O2,5%CO2 and 93% N2). All parasite growth was enhanced by transfer of cultures from a low oxygen to 5% CO2 in air.
With the successful establishment of in vitro cultures for B.equi and B.caballi, in vitro propagated B.equi and B.caballi infected erythrocytes were used as antigen in the IFAT.None showed a positive reaction of a 1 : 80 serum dilution In the serum samples in Japan However, 3 of the 19 trace or false positive ((]SY.+-。[)) cases were observed as pseudo-positive ((]SY.+-。[) -+) in a weak and sharp fluorescence around the parasite of B.equi and/or B.caballi. Antibody survey indicated that both B.equi and B.caballi are endemic in horses in Central Mongolia with 81.8% of horses being seropositive to both infections. Monoclonal antibodies (MAb) were produced against B.equi and B.caballi. One of MAbrecognized 19kDa antigen of B.equi and inhibited the multiplication of B.equi in vitro. This MAb is now being utiuzed to purify the corresponding antigen in order to test as antigen for ELISA.
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