Browsing by Author "Battsetseg, Badgar"

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    The establishment of in vitro culture and drug screening systems for a newly isolated strain of Trypanosoma equiperdum
    (Drugs and Drug Resistance, 2017) Suganuma, Keisuke; Yamasaki, Shino; Innocentia Molefe, Nthatisi; Musinguzi, Peter Simon; Davaasuren, Batdorj; Mossaad, Ehab; Narantsatsral, Sandagdorj; Battur, Banzragch; Battsetseg, Badgar; Inoue, Noboru
    Dourine is caused by Trypanosoma equiperdum via coitus with an infected horse. Although dourine is distributed in Equidae worldwide and is listed as an internationally important animal disease by the World Organization for Animal Health (OIE), no effective treatment strategies have been established. In addition, there are no reports on drug discovery, because no drug screening system exists for this parasite. A new T. equiperdum strain was recently isolated from the genital organ of a stallion that showed typical symptoms of dourine. In the present study, we adapted T. equiperdum IVM-t1 from soft agarose media to HMI-9 liquid media to develop a drug screening assay for T. equiperdum. An intracellular ATP-based luciferase assay using CellTiter-Glo reagent and an intracellular dehydrogenase activitybased colorimetric assay using WTS-8 tetrazolium salt (CCK-8 reagent) were used in order to examine the trypanocidal effects of each compound. In addition, the IC50 values of 4 reference trypanocidal compounds (pentamidine, diminazene, suramin and melarsomine) were evaluated and compared using established assays. The IC50 values of these reference compounds corresponded well to previous studies involving other strains of T. equiperdum. The luciferase assay would be suitable for the mass screening of chemical libraries against T. equiperdum because it allows for the simple and rapid-evaluation of the trypanocidal activities of test compounds, while a simple, inexpensive colorimetric assay will be applicable in developing countries for the evaluation of the drug sensitivity of epidemic trypanosome strains.
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    The evaluation of GM6-based ELISA and ICT as diagnostic methods on a Mongolian farm with an outbreak of non-tsetse transmitted horse trypanosomosis
    (Veterinary Parasitology, 2017) Davaasuren, Batdorj; Amgalanbaatar, Tovuu; Musinguzi, Simon Peter; Suganuma, Keisuke; Otgonsuren, Davaajav; Mossaad, Ehab; Narantsatsral, Sandagdorj; Battur, Banzragch; Battsetseg, Badgar; Xuan, Xuenan; Inoue, Noboru
    Trypanosoma equiperdum, which is the etiological agent of dourine, spreads through sexual intercourse in equines. Dourine (T. equiperdum) has been reported in Mongolia, where it is considered an economically important disease of horses. T. evansi has also been reported in Mongolian domestic animals. The objective of this study was to evaluate the potential application of recombinant T. evansi GM6 (rTeGM6-4r)-based diagnostic methods on a farm with an outbreak of non-tsetse transmitted horse trypanosomosis. Ninety-seven percent homology was found between the amino acid sequences of T. equiperdum GM6 and the GM6 of another Trypanozoon, which also shared the same cellular localization. This finding suggests the utility of rTeGM6-4r-based serodiagnostic methods for epidemiological studies and the diagnosis of both surra and dourine in Equidae. Fifty blood samples were examined from a herd of horses. The diagnostic value of an rTeGM6-4r-based ELISA and an rTeGM6-4r-based immunochromatographic test (ICT) were measured in comparison to a T. evansi crude antigen-based ELISA, which is a diagnostic method recommended by the OIE. However, this is not a perfect diagnostic method for trypanosomosis. Positive serum samples were detected in 46%, 42% and 28% of the tested horses using an rTeGM6-4r-based ELISA, crude antigen-based ELISA and rTeGM6-4r-based ICT, respectively. The sensitivity of rTeGM6-based ELISA was 81%, the specificity was 79%, and the agreement was moderate. We conclude that rTeGM6-4r-based ELISA and ICT represent alternative options for baseline epidemiological studies and the on-site diagnosis of horse trypanosomoses in the field, respectively.
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    Isolation, cultivation and molecular characterization of a new Trypanosoma equiperdum strain in Mongolia
    (Parasites & vectors, 2016) Suganuma, Keisuke; Narantsatsral, Sandagdorj; Battur, Banzragch; Yamasaki, Shino; Otgonsuren, Davaajav; Musinguzi, Simon Peter; Davaasuren, Batdorj; Battsetseg, Badgar; Inoue, Noboru
    Trypanosoma equiperdum causes dourine via sexual transmission in Equidae. T. equiperdum is classified under the subgenus Trypanozoon along with the T. brucei sspp. and T. evansi; however, the species classification of Trypanozoon remains a controversial topic due to the limited number of T. equiperdum reference strains. In addition, it is possible that some were misclassified T. evansi strains. Thus, there is a strong need for a new T. equiperdum strain directly isolated from the genital mucosa of a horse with a clinically- and parasitologically-confirmed dourine infection. Methods: Trypanosomes isolated from the urethral tract of a stallion with suspected dourine, were directly cultivated using soft agarose media at 37 °C in 5 % CO2. For molecular characterization, 18S ribosomal RNA (rRNA) gene, the internal transcribed spacer (ITS) and 8 maxicircle DNA regions were amplified by a PCR and their sequences were determined. To analyze the ratio of the kinetoplastic/akinetoplastic population, the kinetoplasts and the nuclei of trypanosomes were subjected to Hoechst staining and observed by fluorescence microscopy. Results: In addition to the clinical symptoms and the molecular diagnosis, this stallion was definitively diagnosed with dourine by the detection of trypanosomes in the urethral mucosa. These results strongly suggested that the isolated trypanosome was true T. equiperdum. T. equiperdum isolated from the urethral tract was adapted in vitro using soft agarose media. Based on the results of a phylogenetic analysis of 18S rRNA and ITS, this T. equiperdum isolate was classified into the Trypanozoon clade. In a PCR of the maxicircle DNA region, only NADH-dehydrogenase subunits 4 and 5 was amplified. Clear kinetoplasts were observed in most of the T. equiperdum isolates. In contrast, most cultureadapted T. equiperdum were of the akinetoplastic form. Conclusion: We concluded that our isolated trypanosome was the first confirmed case of T. equiperdum in Mongolia and named it “T. equiperdum IVM-t1”. T. equiperdum IVM-t1 was well adapted and propagated in soft agarose media, which indicates that this culture method is useful for isolation of T. equiperdum from horses with dourine.
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    A PCR-based survey of animal trypanosomes among domestic animals herded together in the Bayan-Ulgii and Khovd provinces of Mongolia
    (The Journal of Protozoology Research, 2022) Musinguzi, Simon Peter; Suganuma, Keisuke; Davaasuren, Batdorj; Battsetseg, Badgar; Battur, Banzragch; Inoue, Noboru
    Trypanosoma evansi affects a wide range of domestic animal hosts and can be mechanically cross-transmitted by biting flies when various animals graze together. A total of 1,058 animals (camels, yaks, goats, sheep, cattle, and horses), herded together in the Bayan-Ulgii and Khovd provinces of Mongolia, were screened for animal trypanosomosis using KIN (ITS1) PCR; 21.27% of the samples tested positive. There were significant differences in prevalence among animal species (small ruminants, p < 0.01), sex (males, p < 0.05), and age (adult, p < 0.05). Considering the location, a significant difference in sheep was observed, with the prevalence of trypanosomosis being higher among sheep in Khovd Province (p < 0.01) than in Bayan-Ulgii. This is the first study to highlight the prevalence of animal trypanosomosis in domestic animals that range together in Mongolian grasslands. This study also highlights the significance of small ruminants as possible reservoirs of trypanosomes, and shows the relationship between herd structure, age, and sex and trypanosomosis prevalence in Mongolia.