The establishment of in vitro culture and drug screening systems for a newly isolated strain of Trypanosoma equiperdum.
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 activity-based 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 IC values of 4 reference trypanocidal compounds (pentamidine, diminazene, suramin and melarsomine) were evaluated and compared using established assays. The IC 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.
Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.
Publication Date: 2017-04-13 PubMed ID: 28437733PubMed Central: PMC5403793DOI: 10.1016/j.ijpddr.2017.04.002Google Scholar: Lookup
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Summary
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This research article details the development of new in vitro culture and drug screening systems for a strain of Trypanosoma equiperdum, a parasite causing the disease Dourine in horses, and further explores potential treatments.
Introduction to Dourine and T. equiperdum
- Dourine is a disease affecting horses, caused by the parasite Trypanosoma equiperdum. The disease is transmitted through sexual contact with an infected horse.
- Despite the disease’s global prevalence and its classification as an internationally important animal disease by the World Organization for Animal Health, effective treatment strategies have yet to be established.
- This research study focused on a new strain of T. equiperdum isolated from a stallion exhibiting symptoms characteristic of Dourine.
Establishing New In Vitro Culture and Drug Screening Systems
- The researchers adapted the T. equiperdum strain from a soft agarose medium to an HMI-9 liquid medium with the goal of developing a drug screening system.
- They utilized two assays in examining trypanocidal effects of different compounds: an intracellular ATP-based luciferase assay using CellTiter-Glo reagent, and an intracellular dehydrogenase activity-based colorimetric assay using WST-8 tetrazolium salt (CCK-8).
- These assays were specially chosen as they allow for the effective screening of possible agents against T. equiperdum.
Evaluation of Reference Trypanocidal Compounds
- The study evaluated the inhibitory concentration (IC) values of four established trypanocidal compounds: pentamidine, diminazene, suramin, and melarsomine.
- These IC values were compared using the newly-developed assays, and the results reflected well on earlier studies conducted on other T. equiperdum strains.
- The findings highlight the potential applicability of the established reference compounds as treatment for this specific strain of T. equiperdum.
Potential Applications for New Drug Screening Assays
- The luciferase assay is highlighted as being suitable for mass screening of chemical libraries against T. equiperdum. It enables simple and swift evaluation of trypanocidal activities of test compounds.
- The colorimetric assay, described as simple and inexpensive, would be useful in evaluating the drug sensitivity of epidemic trypanosome strains in developing countries.
- These newly-developed assays are steps toward establishing an effective treatment strategy for Dourine.
Cite This Article
APA
Suganuma K, Yamasaki S, Molefe NI, Musinguzi PS, Davaasuren B, Mossaad E, Narantsatsral S, Battur B, Battsetseg B, Inoue N.
(2017).
The establishment of in vitro culture and drug screening systems for a newly isolated strain of Trypanosoma equiperdum.
Int J Parasitol Drugs Drug Resist, 7(2), 200-205.
https://doi.org/10.1016/j.ijpddr.2017.04.002 Publication
Researcher Affiliations
- Research Center for Global Agromedicine, Obihiro University of Agriculture and Veterinary Medicine, Inada, Obihiro, Hokkaido, Japan; National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada, Obihiro, Hokkaido, Japan. Electronic address: k.suganuma@obihiro.ac.jp.
- National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada, Obihiro, Hokkaido, Japan. Electronic address: yamasakishino0824@yahoo.co.jp.
- National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada, Obihiro, Hokkaido, Japan. Electronic address: nthatisimolefe@gmail.com.
- National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada, Obihiro, Hokkaido, Japan. Electronic address: pmusinguzi@yahoo.com.
- National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada, Obihiro, Hokkaido, Japan; Institute of Veterinary Medicine, Laboratory of Molecular Genetics, Mongolian University of Life Sciences, Zaisan 17024, Ulaanbaatar, Mongolia. Electronic address: davlag_mgl@yahoo.com.
- National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada, Obihiro, Hokkaido, Japan; Department of Pathology, Parasitology and Microbiology, Collage of Veterinary Medicine, Sudan University of Science and Technology, P.O. Box 204, Khartoum, Sudan. Electronic address: ehabmssd7@gmail.com.
- Institute of Veterinary Medicine, Laboratory of Molecular Genetics, Mongolian University of Life Sciences, Zaisan 17024, Ulaanbaatar, Mongolia. Electronic address: naran69@gmail.com.
- Institute of Veterinary Medicine, Laboratory of Molecular Genetics, Mongolian University of Life Sciences, Zaisan 17024, Ulaanbaatar, Mongolia. Electronic address: bat912b@yahoo.com.
- Institute of Veterinary Medicine, Laboratory of Molecular Genetics, Mongolian University of Life Sciences, Zaisan 17024, Ulaanbaatar, Mongolia. Electronic address: bata07@gmail.com.
- Obihiro University of Agriculture and Veterinary Medicine, Inada, Obihiro, Hokkaido, Japan. Electronic address: ircpmi@obihiro.ac.jp.
MeSH Terms
- Animals
- Antiprotozoal Agents / pharmacology
- Colorimetry / methods
- Drug Evaluation, Preclinical / methods
- Horse Diseases / parasitology
- Horses
- Inhibitory Concentration 50
- Luminescent Measurements / methods
- Parasitic Sensitivity Tests / methods
- Trypanosoma / drug effects
- Trypanosoma / growth & development
- Trypanosoma / isolation & purification
- Trypanosomiasis / parasitology
- Trypanosomiasis / veterinary
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This article includes 18 references
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Citations
This article has been cited 6 times.- Uchida M, Higashimoto R, Hoshino T, Natori T, Hatanaka D, Hayashi H, Kanaki T, Suzuki T, Ishii I. Reversible suspension culture of human vascular smooth muscle cells using the functional biopolymer FP003. In Vitro Cell Dev Biol Anim 2023 Jul 28;.
- Munsimbwe L, Seetsi A, Namangala B, N'Da DD, Inoue N, Suganuma K. In Vitro and In Vivo Trypanocidal Efficacy of Synthesized Nitrofurantoin Analogs. Molecules 2021 Jun 2;26(11).
- Sanità G, Carrese B, Lamberti A. Nanoparticle Surface Functionalization: How to Improve Biocompatibility and Cellular Internalization. Front Mol Biosci 2020;7:587012.
- Nurbyek S, Murata T, Suganuma K, Ichimaru Y, Kurosawa N, Ishikawa Y, Buyankhishig B, Davaapurev BO, Byambajav T, Otgonsugar P, Sasaki K, Batkhuu J. Isolation of Sesquiterpenoids and Trypanocidal Constituents from Artemisia adamsii. ACS Omega 2025 May 20;10(19):19665-19674.
- Janse van Rensburg HD, N'Da DD, Suganuma K. In Vitro and In Vivo Trypanocidal Efficacy of Nitrofuryl- and Nitrothienylazines. ACS Omega 2023 Nov 14;8(45):43088-43098.
- Janse van Rensburg HD, Suganuma K, N'Da DD. In vitro trypanocidal activities and structure-activity relationships of ciprofloxacin analogs. Mol Divers 2024 Aug;28(4):2667-2680.
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