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. 2018 Nov;30(6):942-945.
doi: 10.1177/1040638718798688. Epub 2018 Sep 11.

Molecular detection of equine trypanosomiasis in the Riyadh Province of Saudi Arabia

Abdullah D Alanazi  1   2   3   4   5   6 , Robert Puschendorf  1   2   3   4   5   6 , Bashir Salim  1   2   3   4   5   6 , Mohamed S Alyousif  1   2   3   4   5   6 , Ibrahim O Alanazi  1   2   3   4   5   6 , Hajri R Al-Shehri  1   2   3   4   5   6
Affiliations

Molecular detection of equine trypanosomiasis in the Riyadh Province of Saudi Arabia

Abdullah D Alanazi et al. J Vet Diagn Invest. 2018 Nov.

Abstract

We conducted a cross-sectional study to detect trypanosome infections of horses and donkeys in the Riyadh Province of Saudi Arabia. DNA was extracted from blood samples collected from 368 horses and 142 donkeys, and subjected to universal first ribosomal internal transcribed spacer region (ITS1)-PCR followed by Trypanosoma evansi species-specific RoTat1.2-PCR. The universal ITS1-PCR revealed T. evansi infection in horses ( n = 12; 3.3%) and donkeys ( n = 4; 2.8%). There was no significant effect of sex or age on the prevalence of trypanosomiasis in horses or donkeys. Application of the RoTat1.2-PCR revealed that the RoTat1.2 VSG gene was absent from the positive ITS1-PCR samples of 3 horses and 1 donkey. This discrepancy could be explained by the circulation of T. evansi type B in Saudi Arabia; however, this suspicion requires confirmation.

Keywords: Equine; ITS1; PCR; RoTat1.2; Saudi Arabia; trypanosomiasis.

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Conflict of interest statement

Declaration of conflicting interests: The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Figures

Figure 1.
Figure 1.
Map of Riyadh Province, Saudi Arabia.
Figure 2.
Figure 2.
Results of the 1.8% agarose gel electrophoresis showing amplified DNA of the 480-bp band that is specific for Trypanosoma brucei subspecies (T. evansi) using ITS1-PCR. Lane M = 100-bp molecular size marker (GeneRuler); lane T = Trypanosoma spp. positive control DNA; lane N = negative PCR control (water); lanes 1–12 = template DNA isolated from horse blood samples; lanes 13–16 = template DNA isolated from donkey blood samples.
Figure 3.
Figure 3.
Results of 1.8% agarose gel electrophoresis separating the amplicons of the RoTat1.2 VSG gene of Trypanosoma evansi. Lane M = 100-bp molecular size marker (GeneRuler); lane T.e = T. evansi positive control DNA; lane N = negative PCR control (water); lanes 1–12 = template DNA isolated from horse blood samples; lanes 13–16 = template DNA isolated from donkey blood samples. All samples were weakly positive except samples 3, 6, 9, and 16, which were negative. The 151-bp band is specific for T. evansi using the TeRoTat920F/TeRoTat1070R primer set.
Figure 4.
Figure 4.
Phylogenetic relationships based on the partial nucleotide sequences (4983-bp) of ITS1 ribosomal DNA of Trypanosoma (Riyadh equine) with other variants of T. evansi in GenBank. The evolutionary history was inferred by using the maximum likelihood method based on the Tamura–Nei model. Evolutionary analyses were conducted in MEGA v.7. Numbers at nodes are bootstrap values derived from 1,000 replicates.
See this image and copyright information in PMC

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