Characterization and phylogenetic analysis of multidrug-resistant protein-encoding genes in Trypanosoma evansi isolated from buffaloes in Ngawi district, Indonesia

Vet World Vol.12 October-2019 Article-9

Research Article

Veterinary World, 12(10): 1573-1577

https://doi.org/10.14202/vetworld.2019.1573-1577

Mohammad Mirza Nuryady¹, Rini Widayanti², Raden Wisnu Nurcahyo³, Brilyantika Fadjrinatha³, and Ahmad Fahrurrozi Z. S.⁴

1. Department of Biology Education, Faculty of Teacher Training and Education, Universitas Muhammadiyah Malang, Malang, Indonesia.
2. Department of Biochemistry, Veterinary Medicine Faculty, Universitas Gadjah Mada, Yogyakarta, Indonesia.
3. Department of Parasitology, Veterinary Medicine Faculty, Universitas Gadjah Mada, Yogyakarta, Indonesia.
4. Department of Tropical Medicine, Medicine Faculty, Universitas Gadjah Mada, Yogyakarta, Indonesia.

Background and Aim: Excessive use of trypanocidal drugs can lead to cases of drug resistance. Multiple cases of resistance have been widely reported for drugs such as isometamidium chloride and diminazene aceturate. These cases deserve serious attention, especially in Indonesia, where the first case was recorded and where the molecular basis of trypanocidal drug resistance has never been evaluated. This study aimed to analyze the multidrug resistance protein (MRP) gene in Trypanosoma evansi isolates, sampled from Indonesia, by focusing on the phylogenetic relationship between these isolates and other Trypanosoma spp.

Materials and Methods: A total of 88 blood samples were drawn from buffaloes in the Ngawi district, Indonesia. Animals infected with T. evansi were detected through the microhematocrit technique and Giemsa blood smear methods. Positive blood samples were used to inoculate in male mice (Mus musculus BALB-C strain) as an animal model for culturing the T. evansi. The genomic DNA of the blood taken from the T. evansi-infected mice was used for polymerase chain reaction amplification, sequencing, and phylogenetic analysis.

Results: Two genes were analyzed; the first gene detected for T. evansi corresponded to Trypanosoma brucei with a homology of 99% and the second gene to Trypanosoma brucei gambiense, with a homology of 100%. These two genes of the MRP from T. evansi showed clear similarity to the MRPE and MRPA genes of the T. brucei ssp.

Conclusion: The MRP gene is conserved on the subspecies level of T. brucei. Only few point mutations were found between various sequences, which mean that the proteins have the same structure. This is important to treat the parasite with the appropriate drugs in the future. Keywords: multidrug-resistant protein gene, phylogenetic analysis, surra, Trypanosoma evansi.

Keywords: multidrug-resistant protein gene, phylogenetic analysis, surra, Trypanosoma evansi.

How to cite this article: Nuryady MM, Widayanti R, Nurcahyo RW, Fadjrinatha B, Fahrurrozi AZS (2019) Characterization and phylogenetic analysis of multidrug-resistant protein-encoding genes in Trypanosoma evansi isolated from buffaloes in Ngawi district, Indonesia, Veterinary World, 12(10): 1573-1577.

Received: 12-04-2019 Accepted: 03-09-2019 Published online: 17-10-2019

Corresponding author: Raden Wisnu Nurcahyo E-mail: wisnu-nc@ugm.ac.id

DOI: 10.14202/vetworld.2019.1573-1577

Copyright: Nuryady, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.