Molecular Characterization, Oxidative Stress-Mediated Genotoxicity, and Hemato-Biochemical Changes in Domestic Water Buffaloes Naturally Infected with <i>Trypanosoma evansi</i> Under Field Conditions
(1) Background: Surra is a debilitating disease of wild and domestic animals caused by <i>Trypanosoma evansi</i> (<i>T. evansi</i>), resulting in significant mortality and production losses in the affected animals. This study is the first to assess the genetic relationships o...
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| author | Waqas Ahmad Muhammad Yasin Tipu Muti ur Rehman Khan Haroon Akbar Aftab Ahmad Anjum Muhammad Ovais Omer |
| author_facet | Waqas Ahmad Muhammad Yasin Tipu Muti ur Rehman Khan Haroon Akbar Aftab Ahmad Anjum Muhammad Ovais Omer |
| author_sort | Waqas Ahmad |
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| description | (1) Background: Surra is a debilitating disease of wild and domestic animals caused by <i>Trypanosoma evansi</i> (<i>T. evansi</i>), resulting in significant mortality and production losses in the affected animals. This study is the first to assess the genetic relationships of <i>T. evansi</i> in naturally affected buffaloes from Multan district, Pakistan, using ITS-1 primers and evaluating the effects of parasitemia and oxidative stress on DNA damage and hematobiochemical changes in infected buffaloes. (2) Methods: Blood samples were collected from 167 buffaloes using a multi-stage cluster sampling strategy, and trypomastigote identification was performed through microscopy and PCR targeting RoTat 1.2 and ITS-1 primers. Molecular characterization involved ITS-1 via neighbor-joining analysis. The impact of parasitemia loads was correlated with oxidative stress markers, genotoxicity, and hematobiochemical parameters using Pearson correlation and multivariable regression models. (3) Results: Field-stained thin blood film microscopy and molecular identification revealed 8.98% and 10.18% infection rates, respectively. Phylogenetic analysis based on ITS-1 region sequences of the identified isolates showed close genetic associations with Indian isolates. The mean trypomastigote count observed in the infected buffaloes was 5.15 × 10<sup>6</sup> (±5.3 × 10<sup>2</sup>)/µL of blood. The parasitemia loads were significantly correlated with the alterations in oxidative stress markers, DNA damage, and changes in hematobiochemical parameters. Infected animals exhibited significant (<i>p</i> < 0.05) alterations in oxidative stress biomarkers, including catalase, nitric oxide, and malondialdehyde concentrations. Noteworthily, a comet assay revealed a significantly (<i>p</i> < 0.0001) higher mean genetic damage index in the infected buffaloes (0.7 ± 0.04) compared with the healthy ones (0.196 ± 0.004). Alongside significant (<i>p</i> < 0.05) reductions in red cell indices, a marked elevation in leukocyte counts and serum hepatic enzyme levels was recorded in the affected buffaloes. (4) Conclusion: <i>T. evansi</i> isolates of buffaloes from Multan, Pakistan, have genetic similarities to Indian isolates. This study also revealed that higher parasitemia loads induce genotoxicity in the infected animals through oxidative stress and cause hematobiochemical alterations under natural field conditions. |
| format | Article |
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| spelling | doaj-art-2b0804d62dc248d48d763c91f4bab8832025-01-24T13:44:47ZengMDPI AGPathogens2076-08172025-01-011416610.3390/pathogens14010066Molecular Characterization, Oxidative Stress-Mediated Genotoxicity, and Hemato-Biochemical Changes in Domestic Water Buffaloes Naturally Infected with <i>Trypanosoma evansi</i> Under Field ConditionsWaqas Ahmad0Muhammad Yasin Tipu1Muti ur Rehman Khan2Haroon Akbar3Aftab Ahmad Anjum4Muhammad Ovais Omer5Department of Pathology, Faculty of Veterinary Sciences, University of Veterinary and Animal Sciences, Lahore 54000, PakistanDepartment of Pathology, Faculty of Veterinary Sciences, University of Veterinary and Animal Sciences, Lahore 54000, PakistanDepartment of Pathology, Faculty of Veterinary Sciences, University of Veterinary and Animal Sciences, Lahore 54000, PakistanDepartment of Parasitology, Faculty of Veterinary Sciences, University of Veterinary and Animal Sciences, Lahore 54000, PakistanInstitute of Microbiology, Faculty of Veterinary Sciences, University of Veterinary and Animal Sciences, Lahore 54000, PakistanDepartment of Pharmacology and Toxicology, Faculty of Biosciences, University of Veterinary and Animal Sciences, Lahore 54000, Pakistan(1) Background: Surra is a debilitating disease of wild and domestic animals caused by <i>Trypanosoma evansi</i> (<i>T. evansi</i>), resulting in significant mortality and production losses in the affected animals. This study is the first to assess the genetic relationships of <i>T. evansi</i> in naturally affected buffaloes from Multan district, Pakistan, using ITS-1 primers and evaluating the effects of parasitemia and oxidative stress on DNA damage and hematobiochemical changes in infected buffaloes. (2) Methods: Blood samples were collected from 167 buffaloes using a multi-stage cluster sampling strategy, and trypomastigote identification was performed through microscopy and PCR targeting RoTat 1.2 and ITS-1 primers. Molecular characterization involved ITS-1 via neighbor-joining analysis. The impact of parasitemia loads was correlated with oxidative stress markers, genotoxicity, and hematobiochemical parameters using Pearson correlation and multivariable regression models. (3) Results: Field-stained thin blood film microscopy and molecular identification revealed 8.98% and 10.18% infection rates, respectively. Phylogenetic analysis based on ITS-1 region sequences of the identified isolates showed close genetic associations with Indian isolates. The mean trypomastigote count observed in the infected buffaloes was 5.15 × 10<sup>6</sup> (±5.3 × 10<sup>2</sup>)/µL of blood. The parasitemia loads were significantly correlated with the alterations in oxidative stress markers, DNA damage, and changes in hematobiochemical parameters. Infected animals exhibited significant (<i>p</i> < 0.05) alterations in oxidative stress biomarkers, including catalase, nitric oxide, and malondialdehyde concentrations. Noteworthily, a comet assay revealed a significantly (<i>p</i> < 0.0001) higher mean genetic damage index in the infected buffaloes (0.7 ± 0.04) compared with the healthy ones (0.196 ± 0.004). Alongside significant (<i>p</i> < 0.05) reductions in red cell indices, a marked elevation in leukocyte counts and serum hepatic enzyme levels was recorded in the affected buffaloes. (4) Conclusion: <i>T. evansi</i> isolates of buffaloes from Multan, Pakistan, have genetic similarities to Indian isolates. This study also revealed that higher parasitemia loads induce genotoxicity in the infected animals through oxidative stress and cause hematobiochemical alterations under natural field conditions.https://www.mdpi.com/2076-0817/14/1/66<i>Trypanosoma evansi</i>genotoxicityITS-1 primeroxidative stressbuffalo healthhematobiochemical markers |
| spellingShingle | Waqas Ahmad Muhammad Yasin Tipu Muti ur Rehman Khan Haroon Akbar Aftab Ahmad Anjum Muhammad Ovais Omer Molecular Characterization, Oxidative Stress-Mediated Genotoxicity, and Hemato-Biochemical Changes in Domestic Water Buffaloes Naturally Infected with <i>Trypanosoma evansi</i> Under Field Conditions Pathogens <i>Trypanosoma evansi</i> genotoxicity ITS-1 primer oxidative stress buffalo health hematobiochemical markers |
| title | Molecular Characterization, Oxidative Stress-Mediated Genotoxicity, and Hemato-Biochemical Changes in Domestic Water Buffaloes Naturally Infected with <i>Trypanosoma evansi</i> Under Field Conditions |
| title_full | Molecular Characterization, Oxidative Stress-Mediated Genotoxicity, and Hemato-Biochemical Changes in Domestic Water Buffaloes Naturally Infected with <i>Trypanosoma evansi</i> Under Field Conditions |
| title_fullStr | Molecular Characterization, Oxidative Stress-Mediated Genotoxicity, and Hemato-Biochemical Changes in Domestic Water Buffaloes Naturally Infected with <i>Trypanosoma evansi</i> Under Field Conditions |
| title_full_unstemmed | Molecular Characterization, Oxidative Stress-Mediated Genotoxicity, and Hemato-Biochemical Changes in Domestic Water Buffaloes Naturally Infected with <i>Trypanosoma evansi</i> Under Field Conditions |
| title_short | Molecular Characterization, Oxidative Stress-Mediated Genotoxicity, and Hemato-Biochemical Changes in Domestic Water Buffaloes Naturally Infected with <i>Trypanosoma evansi</i> Under Field Conditions |
| title_sort | molecular characterization oxidative stress mediated genotoxicity and hemato biochemical changes in domestic water buffaloes naturally infected with i trypanosoma evansi i under field conditions |
| topic | <i>Trypanosoma evansi</i> genotoxicity ITS-1 primer oxidative stress buffalo health hematobiochemical markers |
| url | https://www.mdpi.com/2076-0817/14/1/66 |
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