Molecular characterization and high prevalence of Tritrichomonas foetus in bulls from the North-West Province of South Africa using real - time polymerase chain reaction (PCR) and conventional PCR diagnostics

Vet World Vol.18 December-2025 Article - 34

Research Article

Veterinary World, 18(12): 4128-4144

https://doi.org/10.14202/vetworld.2025.4128-4144

Afaque H. Syed¹, Mpinda Edoaurd Tshipamba², Ngoma Lubanza¹, Baitsholetsi G. Mokolopi², Jean Marie Dibungi Luseba³, and Mulunda Mwanza¹

1. Department of Animal Health, School of Agriculture, Faculty of Natural and Agricultural Sciences, Mafikeng Campus, North-West University, Private Bag X2046, Mmabatho, 2735, South Africa.

2. College of Agriculture and Environmental Sciences, Department of Agriculture and Animal Health, University of South Africa, Florida Science Campus, Johannesburg, South Africa.

3. University of Kinshasa, Faculty of Veterinary Medicine, Democratic Republic of Congo.

Background and Aim: Bovine trichomonosis, caused by Tritrichomonas foetus, is a significant reproductive disease that impacts cattle productivity and breeding efficiency. In South Africa, routine diagnostic methods often depend on culture and microscopy, which may not accurately distinguish T. foetus from nonpathogenic trichomonads. This study aimed to determine the prevalence of T. foetus in bulls from the Dr. Segomotsi Ruth Mompati (DSRM) District, North-West Province, South Africa, using advanced molecular diagnostics, including real-time polymerase chain reaction (RT-PCR), conventional PCR, DNA sequencing, and phylogenetic analysis.

Materials and Methods: A total of 239 sheath wash samples were collected between June 2018 and October 2020. Of these, 51 culture-positive trichomonad isolates were selected for molecular analysis. Microscopy and modified Giemsa staining were used to characterize protozoal morphology. DNA was extracted and subjected to RT-PCR with 5’ TaqMan™ probes, as well as conventional PCR targeting the 5.8S rRNA/Internal Transcribed Spacer (ITS) regions. PCR amplicons were sequenced, and phylogenetic trees were constructed using MEGA (maximum-likelihood, 1,000 bootstrap replicates). Statistical comparisons between diagnostic methods were performed using Chi-square and Cochran’s Q test.

Results: RT-PCR detected T. foetus in 80.4% (41/51) of the culture-positive samples, with most isolates showing low Ct values, indicating strong positivity. Conventional PCR successfully amplified 12 isolates (300–340 bp), all of which were confirmed as T. foetus by sequencing. Phylogenetic analysis showed that the isolates clustered with the Southern African genotype, exhibiting 77%–87% similarity to Namibian strains and were closely related to Australian and Turkish isolates. No significant correlation was found between geographic location and PCR positivity. RT-PCR demonstrated significantly higher sensitivity than conventional PCR (p < 0.05).

Conclusion: This study confirms a high prevalence of T. foetus in bulls in the DSRM district and demonstrates the superior accuracy of molecular diagnostics compared with culture and microscopy. The identification of genotypes closely related to Southern African strains highlights potential transboundary spread. Incorporating PCR-based screening into routine surveillance is essential for accurate diagnosis, minimizing unnecessary culling, and enhancing reproductive herd health. Further longitudinal studies are recommended to assess disease dynamics and inform regional control programs.

Keywords: Tritrichomonas foetus, RT-PCR, conventional PCR, phylogeny, bovine trichomonosis, South Africa.

How to cite this article: Syed, A. H., Tshipamba, M. E., Lubanza, N., Mokolopi, B. G., Luseba, J. M. D., & Mwanza, M. (2025) Molecular characterization and high prevalence of Tritrichomonas foetus in bulls from the North-West Province of South Africa using real - time polymerase chain reaction (PCR) and conventional PCR diagnostics, Veterinary World, 18(12): 4128–4144.

Received: 15-01-2025 Accepted: 22-11-2025 Published online: 27-12-2025

Corresponding author: Mpinda Edoaurd Tshipamba E-mail: edotshipamba@gmail.com, tshipme@unisa.ac.za

DOI: 10.14202/vetworld.2025.4128-4144

Copyright: Syed, 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.