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      Diagnóstico histopatológico e molecular de lesões sugestivas de tuberculose em búfalos abatidos nos municípios de Macapá e Santana, estado do Amapá Translated title: Histopathological and molecular diagnosis of lesions suggestive of tuberculosis in buffaloes slaughtered in the municipalities of Macapá and Santana, Amapá state, Brazil

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          Abstract

          RESUMO: Este estudo teve como objetivo avaliar lesões sugestivas de tuberculose em búfalos abatidos em matadouros oficiais no Estado do Amapá, Brasil, a fim de confirmar o diagnóstico de tuberculose por avaliação histopatológica e molecular. As amostras de tecido de 20 búfalos que apresentavam lesões sugestivas de tuberculose, dos municípios de Macapá e Santana, foram coletadas. As amostras foram divididas em duas partes: uma delas foi fixada em formalina a 10% tamponada e rotineiramente processadas para avaliação histopatológica, coradas pela hematoxilina-eosina e Ziehl-Neelsen; e o outra parte foi usado para Nested-PCR para o complexo de Mycobacterium tuberculosis (CMT) e para Mycobacterium bovis. As lesões macroscópicas sugestivas de tuberculose foram observadas nos pulmões, linfonodos brônquicos, mediastínicos, retrofaríngeos e submandibulares, fígado e pleura. Histopatologicamente, todas as amostras apresentaram lesões sugestivas de tuberculose, caracterizadas por granulomas compostos por grande quantidade de infiltração de células epitelióides, células de Langerhans e linfócitos, margeando um centro necrótico, calcificado ou não, rodeado por cápsula de tecido conjuntivo fibroso. Bacilos álcool-ácido resistentes foram observados nos tecidos de 3/20 (15%) búfalos. Com relação à detecção molecular, 13/20 (65%) bubalinos apresentaram amostras de tecidos positivos: 6 foram positivos nas Nested-PCRs para CMT e M. bovis, um foi positivo apenas na Nested-PCR para CMT, e 6 foram positivos apenas na Nested-PCR para M. bovis. Os resultados deste estudo demonstram a importância de diagnosticar a tuberculose em búfalos na região e apontam para a necessidade de implementar medidas eficazes para controlar e erradicar a enfermidade.

          Translated abstract

          ABSTRACT: This study aimed to evaluate suggestive lesions of tuberculosis in buffaloes slaughtered in official slaughterhouses in the State of Amapá, Brazil, in order to confirm the diagnosis of tuberculosis by histopathological and molecular evaluation. Tissue samples of 20 buffaloes showing lesions suggestive of tuberculosis, from the municipalities of Macapá and Santana, were collected. The samples were divided into two parts: one was fixed in 10% buffered formalin and routinely processed for histopathological evaluation, stained by hematoxylin-eosin and Ziehl-Neelsen; and the other was used for Nested-PCRs for Mycobacterium tuberculosis complex (MTC) and for Mycobacterium bovis. Gross lesions suggestive of tuberculosis were observed in the lungs, bronchial, mediastinic, retropharyngeal and submandibular lymph nodes, liver and pleura. Histopathologically, all samples showed lesions suggestive of tuberculosis, characterized by granulomas composed of large amount of infiltration of epithelioid cells, Langhans cells and lymphocytes, bordering a necrotic core, calcified or not, surrounded by a fibrous connective tissue capsule. Acid-fast bacilli were observed in the tissues of 3/20 (15%) buffaloes. With regards to the molecular detection, 13/20 (65%) buffaloes showed positive tissue samples: 6 were positive both in the MTC and M. bovis Nested-PCRs, one was positive only in the MTC Nested-PCR, and 6 were positive only in the M. bovis Nested-PCR. The results of this study demonstrate the importance of diagnosing TB in buffaloes in the region and point to the requirement to implement effective measures to control and eradicate the disease.

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          Post mortem diagnosis of Mycobacterium bovis infection in cattle.

          A tentative diagnosis of bovine tuberculosis can be made following the macroscopic detection at necropsy of typical lesions. Histo-pathological examination of the lesion may increase the confidence of the diagnosis but bacteriological isolation of Mycobacterium bovis from the lesion is the only way to make a definitive diagnosis. The sensitivity of gross post mortem examination is affected by the method employed and the anatomical sites examined. Careful examination of as few as 6 pairs of lymph nodes, the lungs and the mesenteric lymph nodes can result in 95% of cattle with macroscopic lesions being identified. Although during post mortem inspection of carcases at abattoir all the principle sites where lesions are to be found were examined, this procedure was found to be insensitive for the detection of lesions. To determine the significance of cattle that give a positive reaction in diagnostic tests but do not have visible lesions (NVL), a bacteriological examination is necessary. NVLs may be due to early infection, poor necropsy technique or infection with mycobacteria other than M. bovis. M. bovis was found to survive best in frozen tissue and the tissue preservative, sodium tetraborate, was found to have adverse effects on viability. It was found desirable to use two different culture media for the primary isolation of M. bovis; agar media for rapid growth and egg media for control of contamination. Additional control of contamination was achieved without adversely affecting the viability by treating the specimen before culture with 0.075% hexadecylpyridinium chloride. The addition of CO2 to the incubation atmosphere did not enhance the recovery of M. bovis. Conventional identification of isolates of M. bovis is by biochemical tests and cultural characteristics, but methods employing monoclonal antibodies and DNA probes may be used to obtain a rapid identification.
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            Novel Mycobacterium tuberculosis Complex Pathogen, M. mungi

            Seven outbreaks involving increasing numbers of banded mongoose troops and high death rates have been documented. We identified a Mycobacterium tuberculosis complex pathogen, M. mungi sp. nov., as the causative agent among banded mongooses that live near humans in Chobe District, Botswana. Host spectrum and transmission dynamics remain unknown.
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              Characterization of Mycobacterium orygis as M. tuberculosis Complex Subspecies

              The oryx bacilli are Mycobacterium tuberculosis complex organisms for which phylogenetic position and host range are unsettled. We characterized 22 isolates by molecular methods and propose elevation to subspecies status as M. orygis. M. orygis is a causative agent of tuberculosis in animals and humans from Africa and South Asia.
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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Journal
                pvb
                Pesquisa Veterinária Brasileira
                Pesq. Vet. Bras.
                Colégio Brasileiro de Patologia Animal - CBPA. Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) (Rio de Janeiro, RJ, Brazil )
                0100-736X
                1678-5150
                November 2017
                : 37
                : 11
                : 1198-1204
                Affiliations
                [4] Campo Grande Mato Grosso do Sul orgnameUniversidade Federal de Mato Grosso do Sul Brazil
                [3] Campo Grande Mato Grosso do Sul orgnameEmbrapa Gado de Corte Brazil
                [2] Castanhal Pará orgnameUniversidade Federal do Pará Brazil
                Article
                S0100-736X2017001101198
                10.1590/s0100-736x2017001100003
                9c071476-d4da-4ddb-9ea6-9eb3b0403fbd

                This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

                History
                : 31 January 2017
                : 20 May 2016
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 63, Pages: 7
                Product

                SciELO Brazil


                tuberculose,búfalos,Amapá,bubalinos,Mycobacterium bovis,histopatologia,Complexo Mycobacterium tuberculosis,Nested-PCR,Histopathology,tuberculosis,buffaloes,Brazil,Mycobacterium tuberculosis complex,Histopatológia

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