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Molecular Prevalence of Different Genotypes of Theileria orientalis Detected from Cattle and Water Buffaloes In Thailand OAK
Altangerel, Khukhuu; Sivakumar, Thillaiampalam; Inpankaew, Tawin; Jittapalapong, Sathaporn; Terkawi, Mohamad Alaa; Ueno, Akio; Xuan, Xuenan; Igarashi, Ikuo; Yokoyama, Naoaki; 玄, 学南; 五十嵐, 郁男; 横山, 直明.
Here we report on an epidemiological study regarding the molecular prevalence of different genotypes of Theileria orientalis present among domestic cattle and water buffalo populations bred in Thailand. A phylogenetic analysis based on the parasitic gene encoding a major piroplasm surface protein revealed the presence of 5 genotypes (Types 1, 3, 5, 7, and N-3) in cattle and 7 genotypes (Types 1, 3, 4, 5, 7, N-2, and N-3) in water buffaloes. Types 4, 7, and N-3 of T. orientalis were reported for the first time in water buffaloes. The previously reported C and Thai types from Thailand clustered as types 7 and 6, respectively, in the present analysis. Great similarities were observed among nucleotide sequences of isolates of the same genotype from cattle and...
Ano: 2011 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3592
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Development and evaluation of an enzyme-linked immunosorbent assay using recombinant p23 for the detection of antibodies to Cryptosporidium parvum in cattle OAK
Bannai, Hiroshi; Nishikawa, Yoshifumi; Seo, Jin-yong; Nakamura, Chinatsu; Zhang, Sofa; Kimata, Isao; Takashima, Yasuhiro; Li, Junyou; Igarashi, Ikuo; Xuan, Xuenan; 西川, 義文; 五十嵐, 郁男; 玄, 学南.
An enzyme-linked immunosorbent assay (ELISA) based on the recombinant p23 of Cryptosporidium parvum was established for the detection of antibodies against C. parvum in cattle. The sensitivity and specificity of the ELISA were evaluated with the standard indirect fluorescent antibody test (IFAT) using sporozoites as antigens. Of 77 bovine sera collected from China, 20 (26.0%) were identified as positive by the IFAT. The same samples were tested with the ELISA. The optical densities at 415 nm were compared to the IFAT results and statistically analyzed to designate a provisional cut-off point. As a result, the cut-off point was concluded to be 0.08, which was considered to be the best condition in the light of its sensitivity (80%) and specificity (73.7%)....
Palavras-chave: Cryptosporidium parvum; P23; ELISA; IFAT.
Ano: 2006 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/142
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The Influence of the regulation of Toxoplasma gondii TgMIC2 transgene on host cell infection OAK
Buates, S.; Xuan, Xuenan; Igarashi, Makoto; Sugimoto, Chihiro; Inoue, Noboru; 玄, 学南; 五十嵐, 慎; 井上, 昇.
Toxoplasma gondii microneme protein 2 (TgMIC2), an apically stored adhesin, was shown to be a key participant involved in the initial attachment and invasion to a host cell. In this study, we had established the clonal line of T. gondii tachyzoites which over-expressed TgMIC2 using the tetracycline repressor (TetR)-based inducible gene expression system. The TgMIC2 over-expression significantly reduced tachyzoite propagation in vitro (p < 0.002) and significantly reduced the parasite virulence in mice (p = 0.002). The over-expression of exogenous TgMIC2 caused the increase of mRNA expression level of endogenous TgMIC2. Wild type parasites mainly expressed TgMIC2115 protein, the microneme store and the surface form of TgMIC2. Over-expression of TgMIC2...
Palavras-chave: Toxoplasma gondii; TgMIC2; Over-expression.
Ano: 2008 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/2236
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Serological evidence of infection of Anaplasma and Ehrlichia in domestic animals in Xinjiang Uygur Autonomous Region area, China OAK
Chahan, Bayin; Jian, Zijian; Xuan, Xuenan; Sato, Yukita; Kabeya, Hidenori; Tuchiya, Kotaro; Itamoto, Kazuhito; Okuda, Masaru; Mikami, Takeshi; Maruyama, Soichi; Inokuma, Hisashi; 玄, 学南; 猪熊, 壽.
Serological methods were utilized to detect Anaplasma and Ehrlichia infection in domestic animals in Xinjiang Uygur Autonomous Region, China. By using an indirect immunofluorescence assay (IFA), antibodies that reacted with Anaplasma phagocytophilum and Ehrlichia chaffeensis were detected mainly in ruminants kept on pastureland in Altai, Ili and Kashgar area. Antibody titers up to 1:320 were recorded. These results indicate that ruminants kept in these areas may be infected with some species of Anaplasma and Ehrlichia.
Palavras-chave: Anaplasma; Ehrlichia domestic animals seroepidemiology Xinjiang Uygur Autonomous Region China.
Ano: 2005 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/742
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Toxoplasma gondii: DNA vaccination with genes encoding antigens MIC2, M2AP, AMA1 and BAG1 and evaluation of their immunogenic potential OAK
Dautu, George; Munyaka, Biscah; Carmen, Gabriella; Zhang, Guohong; Omata, Yoshitaka; Xuan, Xuenan; Igarashi, Makoto.
A combination of antigenic regions of microneme proteins have been previously reported as being protective against chronic toxoplasmosis. In this work, we evaluated immune responses induced by immunizing BALB/c and C57BL/6 mice intradermally with plasmid DNA encoding the protein sequences of Toxoplasma gondii AMA1, MIC2, M2AP and BAG1. Mice immunized with the AMA1 gene developed high levels of serum IgG2a and c antibodies as well as cellular immune responses associated with IFN-γ synthesis suggesting a modulated Th1 type of response. Immunization with the AMA1 gene resulted in a partial but significant protection against the acute phase of toxoplasmosis compared to MIC2, M2AP and BAG1 genes. Therefore, the AMA1 gene appears to generate a strong specific...
Palavras-chave: Toxoplasma gondii; DNA; Deoxyribonucleic acid; DNA immunization; MIC2; Microneme protein-2; M2AP; MIC2 associated protein; AMA1; Apical membrane antigen-1; BAG1; Bradyzoite antigen-1; IgG; Immunoglobulin G.
Ano: 2007 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/1031
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TRANSIENT EXPRESSION OF A GREEN FLUORESCENT PROTEIN GENE IN BABESIA BOVIS OAK
Fujii, Kei; Yokoyama, Naoaki; Takabatake, Noriyuki; Suzuki, Hiroshi; Xuan, Xuenan; Igarashi, Ikuo; 横山, 直明; 鈴木, 宏志; 玄, 学南; 五十嵐, 郁男.
In hemoprotozoa,gene transfer technology provides an important tool to aid in the functional study of parasite genes. In this study,a transfer vector containing the enhanced green fluorescent protein (EGFP) gene laid between the Toxoplasma gondii GRAI promoter region and the GRA2 polyA signal region was constructed and transfected into the in vitro culture of Babesia boνis by the electroporation method.On the first and second days post-transfection,clear positive fluorescences were detected in some parasite bodies, indicating the successful expression of the EGFP gene controlled by the GRA-1 promoter in B. bovis. However, the positive parasites were not ubiquitous and finally disappeared until the forth day post-transfection. This is the first time that...
Ano: 2003 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/631
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CLONING AND EXPRESSION OF AN ANTIGEN OF BABESIA GIBSONI IN ESCHERICHIA COLI AND ITS USE FOR THE IMMUNODIAGNOSIS OF CANINE BABESIOSIS OAK
Fukumoto, Shinya; Sekine, Yukiko; Kimbita, Elikira; Huang, Xiaohong; Xuan, Xuenan; Inoue, Noboru; Yokoyama, Naoaki; Igarashi, Ikuo; Fujisaki, Kozo; Sugimoto, Chihiro; Nagasawa, Hideyuki; Mikami, Takeshi; Suzuki, Hiroshi; 福本, 晋也; 玄, 学南; 井上, 昇; 横山, 直明; 五十嵐, 郁男; 鈴木, 宏志.
Palavras-chave: Babesia gibsoni; Babesia canis; CDNA library; ELISA; P30 gene.
Ano: 2002 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/624
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Prime-boost immunization with DNA followed by a recombinant vaccinia virus expressing P50 induced protective immunity against Babesia gibsoni infection in dogs. OAK
Fukumoto, Shinya; Tamaki, Yoh; Okamura, Masashi; Bannai, Hiroshi; Yokoyama, Natsuko; Suzuki, Tomoko; Igarashi, Ikuo; Suzuki, Hiroshi; Xuan, Xuenan.
A heterologous prime-boost immunization regime with priming DNA followed by recombinant vaccinia virus expressing relevant antigens has been shown to induce effective immune responses against several infectious pathogens. In this study, we constructed a recombinant plasmid and vaccinia virus, both of which expressed P50 of Babesia gibsoni, to investigate the immunogenicity and protective efficacy of a heterologous prime-boost immunization against canine babesiosis. The dogs immunized with the prime-boost regime developed a significantly high level of specific antibody against P50 when compared with the control groups, and the antibody level was strongly increased after a booster immunization with a recombinant vaccinia virus. The prime-boost immunization...
Ano: 2007 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/812
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Artesunate, a potential drug for treatment of Babesia infection OAK
Goo, Youn-Kyoung; Terkawi, M. Alaa; Jia, Honglin; Aboge, G. Oluga; Ooka, Hideo; Kim, Suk; Igarashi, Ikuo; Nishikawa, Yoshifumi; Xuan, Xuenan.
The effect of artesunate, a water-soluble artemisinin derivative, against Babesia species, such as Babesia bovis, Babesia gibsoni, and Babesia microti was studied. Cultures of B. bovis and B. gibsoni were treated with 0.26 μM, 2.6 μM, 26 μM, and 260 μM artesunate, and the growth-inhibitory effect was shown in over 2.6 μM artesunate in day 4 and day 3 post-subculture for B. bovis and B. gibsoni, respectively, in dose-dependent manner. In vivo experiment for B. microti, strong inhibition effects were observed in mouse groups treated with over 1.0 mg/kg body weight of artesunate on day 9 and 10 post-infection. These results suggest that artesunate could be a potential drug for Babesia infection.
Palavras-chave: Artesunate; Babesia bovis; Babesia gibsoni; Babesia microti.
Ano: 2010 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/2820
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Expression of recombinant dense granule protein 7 of Neospora caninum and evaluation of its diagnostic potential for canine neosporosis OAK
Hara, Olgga A; Liao, Min; Baticados, Waren; Bannai, Hiroshi; Zhang, Guohong; Zhang, Sofa; Lee, Eung-goo; Nishikawa, Yoshifumi; Claveria, Florencia G.; Igarashi, Makoto; Nagasawa, Hideyuki; Xuan, Xuenan.
Neospora caninum infection is an important disease affecting bovine and canine populations worldwide. The dense granule antigen 7 of N. caninum (NcGRA7) is considered to be an immunodominant antigen. In the present study, the gene encoding truncated NcGRA7 (NcGRA7t) lacking an N-terminal signal peptide was expressed in Escherichia coli, and its diagnostic potential in an enzyme-linked immunosorbent assay (ELISA) was evaluated. The ELISA could clearly discriminate between known N. caninum-positive and -negative sera from dogs. Serum samples randomly collected from dogs in Japan and China were examined for the diagnosis of neosporosis using the ELISA. Twenty-three of 135 samples (17.03%) and 9 of 95 (9.47%) samples from Japan and China, respectively, were...
Palavras-chave: Canine; ELISA; Neospora caninum; NcGRA7.
Ano: 2006 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/995
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CURRENT SERODIAGNOSTIC MEASURES FOR EQUINE BABESIOSIS OAK
Hirata, Haruyuki; Yokoyama, Naoaki; Xuan, Xuenan; Igarashi, Ikuo; 横山, 直明; 玄, 学南; 五十嵐, 郁男.
Palavras-chave: Babesia equi; Babesia caballi; Diagnosis; ELISA; ICT.
Ano: 2004 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/638
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The epidemiological survey for atovaquone resistant related gene of Babesia gibsoni in Japan OAK
Iguchi, Aiko; Soma, Takehisa; Suzuki, Hiroshi; Xuan, Xuenan.
In 73 gDNA samples from Babesia gibsoni-infected dogs, the M121I variant population was measured by using allele-specific real-time PCR. Although the mechanism of atovaquone against B. gibsoni has not been clearly identified, it is reported that the mitochondria cytochrome b gene of the atovaquone-resistant B. gibsoni had a single-nucleotide substitution at nt363 (G to T), which resulted in the substitution of methionine with isoleucine (M121I). In this study, 3/73 samples showed over 5% M121I variant population. Although the M121I variant population is a low percentage, it runs the risk of spreading drug-resistant parasites. It is important to prevent the spread of drug-resistance, so we need to gather information about this at regular intervals.
Palavras-chave: Allele-specific real-time PCR; Babesia gibsoni; Drug resistance; M121I variant population.
Ano: 2016 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/4421
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Kinetoplast DNA and Procyclic Acidic Repetitive Protein A-α Gene of Trypanosoma evansi OAK
Inoue, Noboru; Honzako, Y.; Hirumi, K.; Xuan, Xuenan; Agatsuma, T.; Nagasawa, Hideyuki; Mikami, Takeshi; Hirumi, Hiroyuki.
Palavras-chave: PARP; KDNA; Trypanosoma evansi; T. brucei.
Ano: 1998 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/276
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Characterization of a leucine aminopeptidase from Toxoplasma gondii OAK
Jia, Honglin; Nishikawa, Yoshifumi; Luo, Yuzi; Yamagishi, Junya; Sugimoto, Chihiro; Xuan, Xuenan.
The M17 family leucine aminopeptidase (LAP) hydrolyze amino acids from the N-terminus of peptides. Many LAPs from parasitic protozoa including Plasmmodium, Trypanosoma and Leishmania, have been intensely investigated because of their crucial roles in parasite biology. In this study, a functional recombinant Toxoplasma gondii LAP (rTgLAP) was expressed in Escherichia coli and its enzymatic activity against synthetic substrates for aminopeptidase, as well as the cellular localization was determined. Our results indicated that TgLAP is a functional aminopeptidase in the cytoplasma of T. gondii.
Palavras-chave: Toxoplasma gondii; Leucine aminopeptidase; Enzymatic activity.
Ano: 2010 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/2822
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Identification of a novel gene encoding a secreted antigen 1 of Babesia gibsoni and evaluation of its use in serodiagnosis OAK
Jia, Honglin; Zhou, Jinlin; Ikadai, Hiromi; Matsuu, Aya; Suzuki, Hiroshi; Fujisaki, Kozo; Xuan, Xuenan; 鈴木, 宏志; 五十嵐, 郁男; 玄, 学南.
Serum from a dog immunized with blood plasma from a B. gibsoni-infected dog, putatively containing secreted antigens, was used to screen a cDNA expression library. A novel gene encoding BgSA1 was identified from the isolated clones. The serum raised in mice immunized with the recombinant BgSA1 expressed in Escherichia coli could recognize a native parasite protein with a molecular mass of 59 kDa. Comparing with the previously established ELISA with recombinant P50 as antigen, the ELISA with recombinant BgSA1 as the antigen was more sensitive when they were used to detect field samples. Moreover, a sandwich ELISA with anti-BgSA1 antibodies could detect the circulating BgSA1 in a serial blood plasma from a dog experimentally infected with B. gibsoni. These...
Palavras-chave: Babesia gibsoni; Secreted Antigen 1; Identification; Serodiagnosis.
Ano: 2006 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/1655
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Characterization of a novel thrombospondin-related protein in Toxoplasma gondii OAK
Kawase, Osamu; Nishikawa, Yoshifumi; Bannai, Hiroshi; Matsuo, Tomohide; Xuan, Xuenan.
Toxoplasma gondii is an obligate intracellular protozoan parasite that invades a wide range of host cells. Upon encountering host cells, the parasite releases a large variety of proteins from secretory organelles, such as micronemes, rhoptries and dense granules. The secretion of microneme protein is essential for parasite invasion. We found that a secreted protein with an altered thrombospondin repeat of Toxoplasma gondii (TgSPATR) was a novel microneme protein, which was different from known microneme proteins that carry thrombospondin repeat domains. TgSPATR was secreted in response to an intra-parasitic elevation of Ca2+ and probably secreted during early stages of parasite invasion. Thus, we suggested that TgSPATR, new member of microneme secretory...
Palavras-chave: Ca2+-dependent secretion; Microneme protein; Secreted protein with an alteredthrombospondin repeat; Toxoplasma gondii.
Ano: 2010 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/2819
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Preliminary evaluation of oligomannose-coated liposome vaccines against lethal protozoan infections in mice OAK
Kuboki, Noritaka; Tiwananthagorn, Weerawan; Takagi, Hideaki; Nakayama, Tomoko; Xuan, Xuenan; Inoue, Noboru; Igarashi, Ikuo; Tsujimura, Kunio; Ikehara, Yuzuru; Kojima, Naoya; Yokoyama, Naoaki.
The oligomannose-coated liposome (OML) vaccine is known to induce cellular immunity specific for the encapsulated antigen in immunized mice. In the present study, we preliminarily evaluated the effect of the OML vaccine encapsulating the soluble protozoan lysate of Toxoplasma gondii, Trypanosoma brucei gambiense, or Babesia rodhaini on the corresponding protozoan infections in mice. After the challenge of T. gondii, the OML vaccine group avoided the high mortality resulting from acute infection that was dominantly observed in other control groups. During the infectious course, the development of the T. gondii-specific antibody, which is an indicator of humoral immunity, was constantly controlled at a lower level in the surviving mice of the OML vaccine...
Palavras-chave: Oligomannose-coated liposome (OML); OML; Soluble protozoan antigen; Antibody responses.
Ano: 2007 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/1050
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Identification and characterization of cross-reactive antigens from Neospora caninum and Toxoplasma gondii OAK
Liao, Min; Xuan, Xuenan; Huang, Xiaohong; Shirafuji, Hiroaki; Fukumoto, Shinya; Hirata, Haruyuki; Suzuki, Hiroshi; Fujisaki, Kozo.
Murine monoclonal antibodies (mAbs) against Neospora caninum tachyzoites were produced to identify the cross-reactive antigens between N. caninum and Toxoplasma gondii. Ten mAbs recognizing cross-reactive antigens of both parasites were obtained and tentatively classified into 6 different groups based on their reactivity patterns in an indirect fluorescent antibody test and Western blot analysis. Three mAbs in group I recognized antigens located on the surface of parasites with molecular masses ranging from 28 to 76 kDa; one mAb in group 2 recognized antigens located on interior organelles of parasites with a molecular mass of 50 kDa; one mAb in group 3 recognized antigens located on interior organelles of parasites with molecular masses of 35 kDa and 14...
Palavras-chave: Neospora caninum; Toxoplasma gondii; Cross-reactive antigen; Protein disulfide isomerase; Heat-shock protein; Ribosomal protein 1.
Ano: 2005 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/816
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Identification and functional analysis of a novel mitochondria-localized 2-Cys peroxiredoxin, BbTPx-2, from Babesia bovis OAK
Masatani, Tatsunori; Asada, Masahito; Hakimi, Hassan; Hayashi, Kei; Yamagishi, Junya; Kawazu, Shin-ichiro; Xuan, Xuenan.
Cysteine-based peroxidases, known as peroxiredoxins (Prx) or thioredoxin peroxidases (TPx), are important antioxidant enzymes that prevent oxidative damage caused by reactive oxygen species (ROS). In this study, we identified a novel mitochondrial 2-Cys Prx, BbTPx-2, from a bovine Babesia parasite, B. bovis. BbTPx-2 complementary DNA (cDNA) encodes a polypeptide of 254 amino acid residues. This protein has a mitochondrial targeting peptide at the N-terminus and two conserved cysteine residues of the typical 2-Cys Prx. By using a thiol mixed-function oxidation assay, the antioxidant activity of recombinant BbTPx-2 was revealed, and its antioxidant activity was comparable to that of a cytosolic 2-Cys Prx from B. bovis, BbTPx-1. Notably, we confirmed that...
Palavras-chave: Antioxidant activity; Babasia bovis; Mitochondria; Peroxiredoxin.
Ano: 2016 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/4487
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Construction of Recombinant Feline Herpesvirus Type 1 Expressing Toxoplasma gondii SRS1 OAK
Mishima, Masayuki; Xuan, Xuenan; Takeiri, Akira; Makala, Levi H. C; Igarashi, Ikuo; Fujisaki, Kozo; Nagasawa, Hideyuki; Mikami, Takeshi.
Palavras-chave: Toxoplasma; Vaccine; SRS1; FHV.
Ano: 2000 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/343
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