The establishment of in vitro culture and drug screening systems for a newly isolated strain of Trypanosoma equiperdum
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Date
2017
Journal Title
Journal ISSN
Volume Title
Publisher
Drugs and Drug Resistance
Abstract
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.
Description
Keywords
Colorimetric assay, Drug screening system, Liquid culture, Luciferase assay, Trypanosoma equiperdum
Citation
Suganuma, K., Yamasaki, S., Molefe, N. I., Musinguzi, P. S., Davaasuren, B., Mossaad, E., ... & Inoue, N. (2017). The establishment of in vitro culture and drug screening systems for a newly isolated strain of Trypanosoma equiperdum. International Journal for Parasitology: Drugs and Drug Resistance, 7(2), 200-205. http://dx.doi.org/10.1016/j.ijpddr.2017.04.002