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A PCR Based Survey of Babesia ovata in Cattle from Various Asian, African and South American Countries OAK
SIVAKUMAR, Thillaiampalam; ASADA, Masahito; BATTSETSEG, Badgar; HUANG, Xiaohong; LAN, Dinh Thi Bich; INPANKAEW, Tawin; P. YBAÑEZ, Adrian; ALHASSAN, Andy; THEKISOE, Oriel M. M.; DE MACEDO, Alan Caine Costa; INOKUMA, Hisashi; IGARASHI, Ikuo; YOKOYAMA, Naoaki; 猪熊, 壽; 五十嵐, 郁男; 横山, 直明.
Babesia ovata is a tick-transmitted hemoprotozoan parasite that infects cattle. In our study, bovine blood samples (n=2,034) were collected from 10 different countries (Brazil, China, Ghana, Japan, Mongolia, the Philippines, South Africa, Sri Lanka, Thailand and Vietnam) and DNA extracted. The DNA samples were screened using an established and specific polymerase chain reaction (PCR) assay targeting the Apical membrane antigen 1 (AMA-1) gene. Parasite DNA was detected among samples collected from Japan, Mongolia and Thailand. Sequence analyses confirmed that the PCR assay detected only B. ovata AMA-1, and that amplicons from different geographical locations were conserved. Our findings highlight the importance of designing adequate strategies to control B....
Palavras-chave: AMA-1; Babesia ovata; Cattle; Mongolia; Thailand.
Ano: 2013 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/4016
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Babesia bovis Dihydroorotate Dehydrogenase (BboDHODH) is a Novel Molecular Target of Drug for Bovine Babesiosis OAK
KAMYINGKIRD, Ketsarin; CAO, Shinuo; MASATANI, Tatsunori; MOUMOUNI, Paul Franck Adjou; VUDRIKO, Patrick; MOUSA, Ahmed Abd El Moniem; TERKAWI, Mohamad Alaa; NISHIKAWA, Yoshifumi; IGARASHI, Ikuo; XUAN, Xuenan; 西川, 義文; 五十嵐, 郁男; 玄, 学南.
The emergence of drug resistance and adverse side effects of current bovine babesiosis treatment suggest that the search of new drug targets and development of safer and effective compounds are required. This study focuses on dihydroorotate dehydrogenase (DHODH), the fourth enzyme of pyrimidine biosynthesis pathway as a potential drug target for bovine babesiosis. Recombinant Babesia bovis DHODH protein (rBboDHODH) was produced in Escherichia coli and used for characterization and measurement of enzymatic activity. Furthermore, the effects of DHODH inhibitors were evaluated in vitro. The recombinant B. bovis DHODH histidine fusion protein (rBboDHODH) had 42.4-kDa molecular weight and exhibited a specific activity of 475.7 ± 245 Unit/mg, a Km = 276.2 µM for...
Palavras-chave: Atovaquone; Babesia bovis; Chemotherapeutic target agent; Dihydroorotate dehydrogenase; Leflunomide.
Ano: 2014 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3918
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Epidemiology of bovine hemoprotozoa parasites in cattle and water buffalo in Vietnam OAK
WEERASOORIYA, Gayani; SIVAKUMAR, Thillaiampalam; LAN, Dinh Thi Bich; LONG, Phung Thang; TAKEMAE, Hitoshi; IGARASHI, Ikuo; INOUE, Noboru; YOKOYAMA, Naoaki.
A PCR-based survey of hemoprotozoa parasites detected Babesia bigemina, Theileria orientalis and Trypanosoma theileri among cattle and water buffalo in Vietnam, and a new Babesia sp. closely related to Babesia ovata was detected in cattle only. In addition, Theileria annulata and Trypanosoma evansi were not detected in both cattle and water buffalo. Phylogenetic analysis detected T. orientalis MPSP genotypes 3, 5, 7 and N3 in cattle and 5, 7, N1 and N2 in water buffalo. Additionally, water buffalo-derived T. theileri CATL sequences clustered together with a previously reported cattle-derived sequence from Vietnam. This is the first report of a new Babesia sp. in cattle, and T. orientalis MPSP genotype 7 and T. theileri in water buffalo in Vietnam.
Palavras-chave: Cattle; Epidemiology; Hemoprotozoa; Vietnam; Water buffalo.
Ano: 2016 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/4344
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Expression Patterns of the Implantation-associated Genes in the Uterus during the Estrous Cycle in Mice OAK
LEE, Dong-Soo; YANAGIMOTO UETA, Yoshiko; XUAN, Xuenan; IGARASHI, Ikuo; FUJISAKI, Kozo; SUGIMOTO, Chihiro; TOYODA, Yutaka; SUZUKI, Hiroshi; 玄, 学南; 五十嵐, 郁男; 鈴木, 宏志.
The mRNA expression patterns of EGF, HB-EGF, Amphiregulin, EGF receptor, IGF-1, CSF-1, IL-1 alpha, IL-1 beta, IL-1 receptor type 1, IL-1 receptor antagonist, LIF, COX-1, COX-2, Mucin-1, calcitonin, and rat USAG-1 mouse homologue, all of which are involved in the process of conceptus implantation to the endometrium, were examined during the estrous cycle by means of real-time quantitative PCR. COX-2, HB-EGF, LIF, Mucin-1, CSF-1, IL-1 alpha, IL-1 beta, and IL-1 receptor antagonist were temporally regulated during the estrous cycle and highly expressed during the estrous stage. In the case of COX-1, EGF, IGF-1, and EGF receptor, the highest mRNA expression was during the diestrous stage. In contrast, the rat USAG-1 mouse homologue mRNA expression did not...
Palavras-chave: Estrous cycle; Implantation; Mice; Ovarian hormone.
Ano: 2005 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3022
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Genotypic Diversity of Theileria orientalis Detected from Cattle Grazing in Kumamoto and Okinawa Prefectures of Japan OAK
YOKOYAMA, Naoaki; UENO, Akio; MIZUNO, Daisuke; KUBOKI, Noritaka; KHUKHUU, Altangerel; IGARASHI, Ikuo; MIYAHARA, Tokuji; SHIRAISHI, Takashi; KUDO, Ryuta; OSHIRO, Mamoru; ZAKIMI, Satoshi; SUGIMOTO, Chihiro; MATSUMOTO, Kotaro; INOKUMA, Hisashi; 横山, 直明; 五十嵐, 郁男; 松本, 高太郎; 猪熊, 壽.
Theileria orientalis is a benign protozoan species that is widely distributed in Japan, yet sometimes causes serious economic losses in the livestock industry. In this study, we conducted a molecular survey based on genes encoding the major piroplasm surface protein (MPSP) and p23 for T. orientalis detected in cattle grazing in southern areas of Japan, consisting of 2 farms in Kumamoto prefecture (Aso and Kuma districts) and 3 farms in Okinawa prefecture (Ishigaki, Iriomote, and Yonaguni Islands). High prevalence rates of T. orientalis infection were shown in all the cattle populations using the diagnostic MPSP- and p23-PCR assays. Phylogenetic analyses revealed 4 MPSP genotypes and 3 p23 genotypes. Furthermore, MPSP genotype-specific PCR methods were...
Palavras-chave: Cattle; Genotype-specific PCR; Japan; MPSP gene; Theileria orientalis.
Ano: 2011 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3584
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Identification and Characterization of a Trypanosoma congolense 46 kDa Protein as a Candidate Serodiagnostic Antigen OAK
ZHOU, Mo; SUGANUMA, Keisuke; RUTTAYAPORN, Ngasaman; NGUYEN, Thu-Thuy; YAMASAKI, Shino; IGARASHI, Ikuo; KAWAZU, Shin-ichiro; SUZUKI, Yasuhiko; INOUE, Noboru; 五十嵐, 郁男; 井上, 昇.
Trypanosoma congolense is a major livestock pathogen in Africa, causing large economic losses with serious effects on animal health. Reliable serodiagnostic tests are therefore urgently needed to control T. congolense infection. In this study, we have identified one T. congolense protein as a new candidate serodiagnostic antigen. The 46.4 kDa protein (TcP46, Gene ID: TcIL3000.0.25950) is expressed 5.36 times higher in metacyclic forms than epimastigote forms. The complete nucleotide sequences of TcP46 contained an open reading frame of 1,218 bp. Southern blot analysis indicated that at least two copies of the TcP46 gene were tandemly-arranged in the T. congolense genome. The recombinant TcP46 (rTcP46) was expressed in Escherichia coli as a glutathione...
Palavras-chave: Diagnosis; ELISA; Nagana; TcP46; Trypanosoma.
Ano: 2014 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3983
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Involvement of a Host Erythrocyte Sialic Acid Content in Babesia bovis Infection OAK
TAKABATAKE, Noriyuki; OKAMURA, Masashi; YOKOYAMA, Naoaki; OKUBO, Kazuhiro; IKEHARA, Yuzuru; IGARASHI, Ikuo; 横山, 直明; 五十嵐, 郁男.
Host sialic acid (SA) has recently been suggested to play an important role in erythrocyte (RBC) infection by Babesia spp. The present study attempted to further determine the specific type of SAs important in the RBC invasion. Bovine RBC was found to bear abundant α2-3-linked SA residues but not α2-6-linked SA in nature, confirmed by flow cytometric analysis of the neuraminidase (Nm)-treated RBCs. Lectin-blot analyses revealed the removal of α2-3-linked SAs from the 97-, 33-, and 31-kDa bands by the Nm treatment. Addition of the Nm-treated RBCs into an in vitro culture of B. bovis resulted in a decreased population of the parasitized RBCs. The thin smear samples from the cultures were then observed under a confocal laser scanning microscope after staining...
Palavras-chave: Babesia bovis; Erythrocyte; Invasion; Sialic acid.
Ano: 2007 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/4166
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Methylcholanthrene自家誘発腫瘍担癌ラットに対するトキソプラズマ溶解抗原の抗腫瘍効果 OAK
MIYAHARA, Kazuro; YOKOO, Naoya; SAKURAI, Haruhisa; IGARASHI, Ikuo; SAKATA, Yu; YOSHIDA, Yutaka; SAITO, Atsushi; HIROSE, Tsuneo; SUZUKI, Naoyoshi; 宮原, 和郎; 五十嵐, 郁男.
Growth of the tumor autoinduced by 20-methylcholanthrene (MC) in rats was inhibited after administration of Toxoplasma lysate antigen (TLA). The antitumor activity of TLA was most obvious in the early stage of tumoral growth. When TLA was administered to rats before the appearance of tumor, tumor formation was delayed slightly. Histopathological studies revealed dense growths of spindle tumor cells in untreated control rat, while enlarged central necrosis with the infiltration of lymphocytes and neutrophils was apparent in TLA-treated rats. According to the immunohistological examination of tumor tissue with anti-Thy-1 antibody, the rats treated with TLA showed large Thy-1 positive granular cells, whereas the untreated rats indicated only a few small Thy-1...
Palavras-chave: Antitumor activity; Methylcholanthrene-autoinduced tumor; Rat; Thy-1 positive granular cell; Toxoplasma lysate antigen.
Ano: 1992 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/4064
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Molecular Epidemiological Survey of Theileria orientalis in Thua Thien Hue Province, Vietnam OAK
KHUKHUU, Altangerel; LAN, Dinh Thi Bich; LONG, Phung Thang; UENO, Akio; LI, Yan; LUO, Yuzi; MACEDO, Alan Caine Costa de; MATSUMOTO, Kotaro; INOKUMA, Hisashi; KAWAZU, Shin-Ichiro; IGARASHI, Ikuo; XUAN, Xuenan; YOKOYAMA, Naoaki; 松本, 高太郎; 猪熊, 壽; 河津, 信一郎; 五十嵐, 郁男; 玄, 学南; 横山, 直明.
Theileria orientalis is a benign bovine protozoan parasite that occasionally causes serious economic loss in the livestock industry. We report the findings of a molecular epidemiological survey of T. orientalis in 94 Vietnamese yellow cattle, 43 water buffaloes, 21 sheep, 21 goats and 85 blood-sucking ticks of cattle in the Thua Thien Hue province of Vietnam. The major piroplasm surface protein (MPSP) gene of T. orientalis was detected using polymerase chain reaction from 13 cattle (13.8%), 11 water buffaloes (25.6%), 1 sheep (4.8%) and 9 ticks (10.6%). Phylogenetic analysis using MPSP gene sequences showed the presence of seven genotypes, four previously categorized genotypes (Types 1, 3, 5 and 7) and three new genotypes (Types N-1, N-2 and N-3).
Palavras-chave: Cattle; Genotype; MPSP; Theileria orientalis; Vietnam.
Ano: 2011 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3585
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PCR Detection and Genetic Diversity of Bovine Hemoprotozoan Parasites in Vietnam OAK
SIVAKUMAR, Thillaiampalam; LAN, Dinh Thi Bich; LONG, Phung Thang; YOSHINARI, Takeshi; TATTIYAPONG, Muncharee; GUSWANTO, Azirwan; OKUBO, Kazuhiro; IGARASHI, Ikuo; INOUE, Noboru; XUAN, Xuenan; YOKOYAMA, Naoaki; 井上, 昇; 玄, 学南; 横山, 直明.
Hemoprotozoan infections often cause serious production losses in livestock. In the present study, we conducted a PCR-based survey of Babesia bovis, Babesia bigemina, Theileria annulata, Theileria orientalis, Trypanosoma evansi and Trypanosoma theileri, using 423 DNA samples extracted from blood samples of cattle (n=202), water buffaloes (n=43), sheep (n=51) and goats (n=127) bred in the Hue and Hanoi provinces of Vietnam. With the exception of T. annulata and T. evansi, all other parasite species (B. bovis, B. bigemina, T. orientalis and T. theileri) were detected in the cattle populations with B. bovis being the most common among them. Additionally, four water buffaloes and a single goat were infected with B. bovis and B. bigemina, respectively. The Hue...
Palavras-chave: Epidemiology; Hemoprotozoa; Livestock; PCR; Vietnam.
Ano: 2013 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/4015
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The Genetic Diversity of Merozoite Surface Antigen 1 (MSA-1) among Babesia bovis Detected from Cattle Populations in Thailand, Brazil and Ghana OAK
NAGANO, Daisuke; SIVAKUMAR, Thillaiampalam; DE DE MACEDO, Alane Caine Costa; INPANKAEW, Tawin; ALHASSAN, Andy; IGARASHI, Ikuo; YOKOYAMA, Naoaki; 五十嵐, 郁男; 横山, 直明.
In the present study, we screened blood DNA samples obtained from cattle bred in Brazil (n=164) and Ghana (n=80) for Babesia bovis using a diagnostic PCR assay and found prevalences of 14.6% and 46.3%, respectively. Subsequently, the genetic diversity of B. bovis in Thailand, Brazil and Ghana was analyzed, based on the DNA sequence of merozoite surface antigen-1 (MSA-1). In Thailand, MSA-1 sequences were relatively conserved and found in a single clade of the phylogram, while Brazilian MSA-1 sequences showed high genetic diversity and were dispersed across three different clades. In contrast, the sequences from Ghanaian samples were detected in two different clades, one of which contained only a single Ghanaian sequence. The identities among the MSA-1...
Palavras-chave: Babesia bovis; Brazil; Ghana; MSA-1; Thailand.
Ano: 2013 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/4014
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トキソブラズマ溶解抗原(TLA)によって誘導される細胞障害性における脾臓の付着性細胞の役割 OAK
MIYAHARA, Kazuro; TOSE, Shinichi; SAKURAI, Haruhisa; IGARASHI, Ikuo; SAITO, Atsushi; HIROSE, Tsuneo; SUZUKI, Naoyoshi; 宮原, 和郎; 五十嵐, 郁男.
In order to identify mechanisms responsible for the anti-tumor effects of Toxoplasma lysate antigen (TLA), we used an in vitro 51Cr release assay to study the functional properties of plastic-adherent cells during induction of splenic cytotoxic activity by TLA. Cytotoxic activity of non-adherent cells was measured in all experiments after a 6 days incubation. Induction of cytotoxic non-adherent cells by TLA required the presence of plastic-adherent spleen cells. In contrast, rhIL-2 alone was able to induce transformation of cytotoxic non-adherent cells from non-adherent spleen cells. Contact between adherent and non-adherent spleen cells was necessary for successful induction of cytotoxic non-adherent cells by TLA. Treatment of spleen cells with...
Palavras-chave: Adherent cell; Cytotoxic activity; Macrophage; Spleen cell; Toxoplasma lysate antigen.
Ano: 1992 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/4065
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トキソプラズマ溶解抗原(TLA)で誘導されるマウス脾臓内の細胞障害性細胞の性質 OAK
MIYAHARA, Kazuro; MATSUMOTO, Toru; TOSE, Shinichi; SAKURAI, Haruhisa; IGARASHI, Ikuo; SAITO, Atsushi; HIROSE, Tsuneo; SUZUKI, Naoyoshi; 宮原, 和郎; 五十嵐, 郁男.
Spleen cells from Toxoplasma lysate antigen (TLA)-sensitized BALB/c mice showed the strong cytotoxic activity against both natural killer (NK)-sensitive cells (YAC-1 and RL♂-1) and NK-insensitive cells (P-815), when incubated with TLA or recombinant human IL-2 (rhIL-2). The increment of TLA concentration in culture medium increased the cytotoxic activity. Treatment of effector cells; spleen cells from TLA-sensitized mice incubated with TLA, with anti-asialo GM1 or anti-Thy-1 plus complement inhibited the cytotoxic activity of effector cells, whereas treatment with anti-mouse Lyt-2.2 serum plus complement had no effect on the cytotoxic activity. Treatment of spleen cells from TLA-sensitized mice with anti-asialo GM1 and/or anti-Thy-1 plus complement...
Palavras-chave: Cytotoxic activity; Mouse; Phenotype; Spleen cell; Toxoplasma lysate antigen.
Ano: 1992 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/4063
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