Bovine Tuberculosis Testing in Colombia: Comparative Histopathological, Microbiological, and Molecular Biology Findings

Paula Palomino Cadavid, D. Balvin, Rafael Villarreal Julio, Enderson Murillo Ramos, Jesus Berdugo Gutierrez, Jhon Didier Ruiz Buitrago, Rene Ramirez Garcia
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Abstract

Introduction: Bovine tuberculosis (bTB) is a zoonotic infectious disease present in Colombia, caused by Mycobacterium bovis, and causes tuberculosis in water buffalo (Bubalus bubalis). Diagnosis of bovine tuberculosis through the intradermal test is difficult; evaluating and understanding the behavior of other diagnostic tests is necessary. Objective: To describe the behavior and results of different diagnostic methods for bovine tuberculosis in water buffalo positive for the Purifed Proteic Derivate (DPP) intradermal test. Methodology: In water buffaloes positive for comparative cervical tuberculin test, different diagnostic methods were applied, described, and compared: Ziehl-Neelsen staining, microbiological culture, histopathological analysis, and PCR-HRM. Results: Histopathological tests showed that 26 water buffalo positive for DPP (52%) had histological lesions compatible with bovine tuberculosis. 37% of the evaluated samples from tuberculin-positive Buffalo's lungs and secondary lymph nodes showed acid-alcohol-resistant bacillus with Ziehl-Neelsen staining. Four samples of Mycobacterium bovis from tuberculin-positive buffalo were isolated and identified, with two of these isolates confirmed from tissues with PCR-HRM, and three buffalo with microbiological isolates presented granulomatous lesions through histological analysis. Seventeen tuberculin-positive buffalo (34%) tested positive for real-time PCR HRM, and nine of these buffalo did not have histological lesions compatible with bTB and were confirmed with the molecular test. Conclusion: Our results provide positive evidence of histological findings, microbiological isolation, and molecular diagnosis of tuberculin-positive water buffalo in the lowlands of Colombia. None of the complementary tests performed showed 100% concordance with the comparative cervical tuberculin test results for bTB.
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哥伦比亚的牛结核病检测:组织病理学、微生物学和分子生物学的比较研究结果
导言:牛结核病(bTB)是一种存在于哥伦比亚的人畜共患传染病,由牛分枝杆菌引起,并导致水牛(Bubalus bubalis)结核病。通过皮内试验诊断牛结核病非常困难,因此有必要评估和了解其他诊断试验的行为。目的描述纯化蛋白衍生物(DPP)皮内试验阳性水牛的牛结核病不同诊断方法的行为和结果。方法:对宫颈结核菌素比较试验呈阳性的水牛采用不同的诊断方法,对其进行描述和比较:Ziehl-Neelsen 染色法、微生物培养法、组织病理学分析法和 PCR-HRM 法。结果组织病理学检测显示,26 头 DPP 阳性水牛(52%)的组织病变与牛结核病相符。在结核菌素阳性水牛的肺部和次级淋巴结的评估样本中,37%的样本经齐氏-奈尔森染色显示出耐酸性杆菌。从结核菌素阳性水牛身上分离并鉴定出了四份牛分枝杆菌样本,其中两份样本通过 PCR-HRM 从组织中得到了证实,三份带有微生物分离物的水牛通过组织学分析出现了肉芽肿病变。17 头结核菌素阳性水牛(34%)的实时 PCR HRM 检测结果呈阳性,其中 9 头水牛的组织学病变与牛结核病不相符,但通过分子检测得到了确认。结论我们的研究结果为哥伦比亚低地结核菌素阳性水牛的组织学发现、微生物分离和分子诊断提供了积极的证据。所进行的辅助检测均未显示与宫颈结核菌素检测结果100%吻合。
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