Abstract
Trypanosoma (Megatrypanum) theileri is a cosmopolitan, usually non-pathogenic, trypanosome of cattle transmitted by blood-sucking arthropods, mainly tabanid flies. Several T. theileri strains isolated from domestic and wild ruminants via co-culturing with mammalian feeder cells or blood cells have been characterized morphologically and genetically. Here, we cultured a new trypanosome isolate from a Holstein cow in Hokkaido, Japan, and performed morphological and molecular characterization studies. The new isolate (Obihiro strain) was co-cultivated with Madin–Darby bovine kidney (MDBK) cells in GIT medium supplemented with 10% fetal bovine serum. Trypomastigotes and epimastigotes, but not intracellular parasites, were identified in the culture. Analysis of the V7-V8 region of 18S rRNA sequences showed that the Obihiro strain is positioned within the subgenus Megatrypanum. A dendrogram based on whole internal transcribed spacer rDNA sequence showed that the Obihiro strain clustered in the lineage TthII together with the Japanese isolates of T. theileri, Esashi 9, and Esashi 12, and isolates from Zambia and the USA. T. theileri of the KM strain and a T. theileri-like trypanosome isolated from deer (TSD1 strain) clustered in the lineage TthI, separate from the Obihiro strain. Based on a partial cathepsin L-like protein gene analysis, the Obihiro strain clustered with isolates of the TthIIF genotype, which includes T. theileri from Vietnam, Sri Lanka, and Brazil. Our analyses of the T. theileri Obihiro strain provide relevant insights into its genetic diversity in Japanese cattle and corroborate the host specificity of cattle and deer trypanosomes of the subgenus Megatrypanum.
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Acknowledgements
We thank Ms. Yoko Matsushita for her excellent technical assistance. This study was partially supported by the “International Collaborative Research Program for Tackling the NTDs (Neglected Tropical Diseases) Challenges in African Countries” from the Japan Agency for Medical Research and Development, AMED. The study was financially supported by the Japan Society for the Promotion of Science (JSPS), KAKENHI Grant Number 16K18793 (Grants-in-Aid for Young Scientists (B)) and the AMED/JICA SATREPS (17jm0110006h0005).
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Supplementary fig. S1
PCR products of Trypanosoma theileri Obihiro strain. All of the PCR-amplified target loci had expected sizes. M: 100 bp marker; Lane 1: V7-V8 region of 18S rRNA; Lane 2: whole internal transcribed spacer (ITS); Lane 3: Tth625 fragment; Lane 4: partial CATL; and Lane 5: ITS1. (PDF 59 kb)
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Suganuma, K., Kondoh, D., Sivakumar, T. et al. Molecular characterization of a new Trypanosoma (Megatrypanum) theileri isolate supports the two main phylogenetic lineages of this species in Japanese cattle. Parasitol Res 118, 1927–1935 (2019). https://doi.org/10.1007/s00436-019-06313-x
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DOI: https://doi.org/10.1007/s00436-019-06313-x