ABSTRACT: Bovine tuberculosis is a zoonotic disease with global distribution. This study aimed to describe its prevalence in cattle through a systematic review and meta-analysis of studies conducted all around the world. The research consisted of a systematic literature review following the precepts of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) methodology. Cross-sectional studies that described the prevalence of the disease using the diagnostic method through an intradermal test with bovine and avian purified protein derivative (PPD) were selected. Identifying articles was performed in the PubMed, ScienceDirect, Scopus, and Web of Science databases. Of the total number of studies found (n=1,839), 60 met all criteria and were included in this review. The overall prevalence was 3.27% (2.11-5.05%) for animals and 18.09% (11.20-27.90%) for herds. Analysis of risk factors for tuberculosis in cattle was found or performed in 50 studies. The heterogeneity identified among the works included was expected, given the differences in research design, year of publication, and the number of animals sampled. It is necessary to evaluate the insertion of new ante mortem diagnostic tests into control and eradication programs, which, combined with allergic tests, may identify the largest number of animals that presented an actual positive for the disease. ABSTRACT distribution metaanalysis meta analysis world MetaAnalyses Meta Analyses PRISMA (PRISMA methodology Crosssectional Cross sectional PPD (PPD selected PubMed ScienceDirect Scopus databases n=1,839, n1839 n n=1,839 , 1 839 (n=1,839) 6 327 3 27 3.27 2.115.05% 211505 2.11 5.05% 2 11 5 05 (2.11-5.05% 1809 18 09 18.09 11.2027.90% 11202790 11.20 27.90% 20 90 (11.20-27.90% herds expected design publication sampled programs which n183 n=1,83 83 (n=1,839 32 3.2 115 2.115.05 21150 211 2.1 505 5.05 0 (2.11-5.05 180 18.0 2027 11.2027.90 1120279 1120 11.2 2790 27.90 9 (11.20-27.90 n18 n=1,8 8 (n=1,83 3. 2.115.0 2115 21 2. 5.0 (2.11-5.0 18. 202 11.2027.9 112027 112 11. 279 27.9 (11.20-27.9 n1 n=1, (n=1,8 2.115. 5. (2.11-5. 11.2027. 11202 27. (11.20-27. n=1 (n=1, 2.115 (2.11-5 11.2027 (11.20-27 n= (n=1 (2.11- 11.202 (11.20-2 (n= (2.11 (11.20- (n (2.1 (11.20 (2. (11.2 (2 (11. ( (11 (1

RESUMO: A tuberculose bovina é uma doença zoonótica com distribuição global. O objetivo do presente trabalho foi descrever a prevalência da enfermidade em bovinos através de uma revisão sistemática e meta-análise em estudos realizados no mundo. A pesquisa consistiu em uma revisão sistemática de literatura seguindo os preceitos da metodologia “Preferred Reporting Items for Systematic Reviews and Meta-Analyses” (PRISMA). Estudos transversais que descreveram a prevalência da enfermidade utilizando o método de diagnóstico através da prova intradérmica com derivado proteico purificado (PPD) bovino e aviário foram selecionados. O processo de identificação dos artigos foi desenvolvido nas bases de dados PubMed, ScienceDirect, Scopus e Web of Science. Do total de estudos encontrados (n=1.839), 60 atenderam a todos os critérios e foram incluídos na presente revisão. A prevalência geral encontrada foi de 3,27% (2,11-5,05%) para animais e 18,09% (11,20-27,90%) para rebanhos. Análises de fatores de riscos para a tuberculose em bovinos foi encontrada ou realizada em 50 estudos. A heterogeneidade identificada entre os trabalhos incluídos nesta revisão era esperada tendo em vista as diferenças relacionadas aos desenhos de pesquisa, anos de publicação dos estudos e a quantidade de animais amostrados. É necessária a avaliação da introdução de novos testes de diagnóstico ante-mortem aos programas de controle e erradicação que, combinados com os testes alérgicos, possam identificar o maior número de animais verdadeiramente positivos para a enfermidade. RESUMO global metaanálise meta análise mundo Preferred MetaAnalyses Meta Analyses Meta-Analyses PRISMA. PRISMA . (PRISMA) PPD (PPD selecionados PubMed ScienceDirect Science n=1.839, n1839 n n=1.839 , 1 839 (n=1.839) 6 327 3 27 3,27 2,115,05% 211505 2,11 5,05% 2 11 5 05 (2,11-5,05% 1809 18 09 18,09 11,2027,90% 11202790 11,20 27,90% 20 90 (11,20-27,90% rebanhos amostrados antemortem ante mortem alérgicos (PRISMA n183 n=1.83 83 (n=1.839 32 3,2 115 2,115,05 21150 211 2,1 505 5,05 0 (2,11-5,05 180 18,0 2027 11,2027,90 1120279 1120 11,2 2790 27,90 9 (11,20-27,90 n18 n=1.8 8 (n=1.83 3, 2,115,0 2115 21 2, 5,0 (2,11-5,0 18, 202 11,2027,9 112027 112 11, 279 27,9 (11,20-27,9 n1 n=1. (n=1.8 2,115, 5, (2,11-5, 11,2027, 11202 27, (11,20-27, n=1 (n=1. 2,115 (2,11-5 11,2027 (11,20-27 n= (n=1 (2,11- 11,202 (11,20-2 (n= (2,11 (11,20- (n (2,1 (11,20 (2, (11,2 (2 (11, ( (11 (1

ABSTRACT: This study determined the frequency of positive tests for bovine tuberculosis in animals and breeders/establishments in Rio Grande do Norte, Brazil. Data were provided by the Instituto de Defesa e Inspeção Agropecuária do Rio Grande do Norte (IDIARN), from its Unidade Local de Sanidade Animal e Vegetal (ULSAV’S), acquired in monthly reports issued by qualified Veterinarians under the National Program for the Control and Eradication of Brucellosis and Animal Tuberculosis (PNCEBT), from June 2012 to December 2021. For diagnosis, the comparative cervical test was used as a routine and confirmatory test. A total of 45,804 cattle were tested, 53 (0.1%) of which tested positive. Secondary data are essential in the evaluation of sanitary measures, allowing rapid generation of hypotheses about diseases and providing support for decision-making. Measures such as raising awareness among producers, sanitary control in the acquisition and sale of cows for reproduction, inspection of sanitary barriers, and conducting epidemiological surveys to understand the actual situation of this disease in the state of Rio Grande do Norte are crucial. ABSTRACT breedersestablishments breeders establishments Brazil IDIARN, IDIARN , (IDIARN) ULSAVS, ULSAVS ULSAV S (ULSAV’S) PNCEBT, PNCEBT (PNCEBT) 201 2021 diagnosis 45804 45 804 45,80 5 0.1% 01 0 1 (0.1% measures decisionmaking. decisionmaking decision making. making decision-making producers reproduction barriers crucial (IDIARN (ULSAV’S (PNCEBT 20 202 4580 4 80 45,8 0.1 (0.1 2 458 8 45, 0. (0. (0 (

RESUMO: O objetivo do trabalho foi determinar a frequência de animais e estabelecimentos positivos para a tuberculose bovina no estado do Rio Grande do Norte, Brasil. Os dados foram fornecidos pelo Instituto de Defesa e Inspeção Agropecuária do Rio Grande do Norte (IDIARN) e são oriundos de suas Unidades Locais de Sanidade Animal e Vegetal (ULSAV) coletados dos relatórios mensais emitidos por Médicos Veterinários habilitados para atuação no âmbito do Programa Nacional de Controle e Erradicação da Brucelose e da Tuberculose Animal (PNCEBT), compreendendo o período de junho de 2012 a dezembro de 2021. Para o diagnóstico foi utilizado o Teste Cervical Comparativo (TCC) como teste de rotina e confirmatório. No total, foram testados 45.804 bovinos, dos quais 53 animais (0,1%) apresentaram resultado positivo. Os dados secundários são informações de extrema importância na avaliação de medidas sanitárias, permitindo uma rápida geração de hipóteses sobre as enfermidades e dando suporte na tomada de decisões. É fundamental a condução de medidas que incluam a conscientização dos produtores, controle sanitário na aquisição e venda de matrizes, fiscalização nas barreiras sanitárias e a realização de um inquérito epidemiológico para se conhecer a real situação desta doença no estado do Rio Grande do Norte. RESUMO Brasil IDIARN (IDIARN ULSAV (ULSAV PNCEBT, PNCEBT , (PNCEBT) 201 2021 TCC (TCC confirmatório total 45804 45 804 45.80 bovinos 5 0,1% 01 0 1 (0,1% positivo decisões produtores matrizes (PNCEBT 20 202 4580 4 80 45.8 0,1 (0,1 2 458 8 45. 0, (0, (0 (

The current study was designed to evaluate the reliability of the interferon-gamma release assay (IGRA) as a screening test compared to intensive post-mortem inspection using agreement analyses. This study also aimed to determine the prevalence and risk factors of bovine tuberculosis (bTB) among bovines presented for slaughter in the largest abattoir in Lahore, Pakistan. After anti-mortem inspection, a total of 102 animals were randomly selected for sample and data collection. Selected animals were slaughtered and a thorough post-mortem examination was done for all carcasses to find TB-like lesions. Blood samples were processed by IGRA. Prevalence estimates were generated and Cohen's Kappa test was done for agreement analyses to compare the reliability of the two tests for bTB diagnosis. A substantial agreement (κ = 0.79) was estimated between the IGRA and intensive post-mortem inspection. The apparent prevalence was computed as 5.88% (95% CI; 2.59-11.97) and the true prevalence was estimated as 3.92% (95% CI; 1.35-9.47). A parallel testing strategy with IGRA and intensive post-mortem inspection is a useful approach for screening bTB.

Resumen Tenosinovitis es la inflamación o infección de un tendón y su vaina sinovial. Una de las causas puede ser bacteriana. Los gérmenes comunes son los más frecuentes, pero deben investigarse también las micobacterias. En la Argentina, la tuberculosis es un problema de salud pública. Mycobacterium bovis ocasiona zoonosis que afectan animales y humanos. La infección en el hombre se da por inhalación de aerosoles de ganado enfermo, por vía digestiva o cutánea. Las manifestaciones extrapulmonares pueden ser digestivas, ganglionares, cutáneas u osteoarticulares. La tenosinovitis tuberculosa de muñeca es infrecuente y, generalmente, es diagnosticada como tenosinovitis reumática o inespecífica. Se presenta el caso de una mujer de 48 años, residente de zona rural, que presentaba hipertrofia y edema en dedos de la mano izquierda. Se le realizó cirugía del túnel carpiano y se observaron cuerpos como granos de arroz. Del tejido sinovial se aisló M. bovis. Comenzó tratamiento tuberculostático más claritromicina. Evolucionó favorablemente.

Resumo A tenossinovite é a inflamação ou infecção de um tendão e sua bainha sinovial. A causa pode ser bacteriana, sendo os germes comuns os mais frequentes, mas as micobactérias devem ser investigadas. Na Argentina, a tuberculose é um problema de saúde pública. Mycobacterium bovis causa zoonose que afeta animais e humanos. A infecção no homem ocorre por inalação de aerossóis de gado doente, por via digestiva ou cutânea. As manifestações extrapulmonares podem ser digestivas, ganglionares, cutâneas, osteoarticulares. A tenossinovite tuberculosa do pulso é pouco frequente, geralmente diagnosticada como tenossinovite reumática ou inespecífica. Apresenta-se o caso de uma mulher de 48 anos, moradora de zona rural, com hipertrofia e edema nos dedos da mão esquerda. Foi realizada cirurgia do túnel do carpo, observando-se corpos como grãos de arroz. Do tecido sinovial foi isolado M. bovis. Iniciou-se tratamento tuberculostático mais claritromicina. Evoluiu favoravelmente.

Abstract Tenosynovitis is an inflammation or infection of a tendon and its synovial sheath. One cause of this illness may be bacterial, being common organisms more frequent, but mycobacteria should be also investigated. In Argentina, tuberculosis is a public health problem. Mycobacterium bovis causes zoonoses that affect animals and humans. Infection in man occurs by inhalation of aerosols from sick cattle, by digestive or cutaneous routes. Extrapulmonary manifestations can be digestive, nodal, cutaneous or osteoarticular. Tuberculous wrist tenosynovitis is uncommon, usually diagnosed as rheumatic or nonspecific tenosynovitis. The case of a 48-year-old woman, a resident of rural areas, presenting hypertrophy, and edema in the fingers of the left hand was introduced. Carpal tunnel surgery was performed, while rice grain bodies were noticed. Synovial tissue material was taken for mycobacterial culture, where M. bovis was isolated. She began tuberculostatic treatment plus clarithromycin. She evolved favourably.

ABSTRACT: Bovine tuberculosis (bTB) is a chronic disease of animals mainly caused by Mycobacterium bovis, a zoonotic pathogen that generates economic losses in the milk and meat industry. In central Chile, the Metropolitan Region concentrates dairy herds with the highest bTB prevalence of the country and the official veterinary service has supported the evaluation of the M. bovis Bacillus Calmette-Guerin (BCG) vaccine in this area with the replacement of tuberculin purified protein derivative (PPDs) by the DIVA (Differentiating Infected from Vaccinated Animals) peptides for the bTB diagnosis in the herds. This study aimed to describe the IFN-ℽ response against PPDs (bovine and avian PPD) and DIVA antigenic cocktails (ESAT-6/CFP-10 and Rv3615c) in BCG vaccinated 11-month-old heifers under a natural transmission scenario. Sixty-two animals were vaccinated via subcutaneous route with a 2-8 x 105 colony forming units of BCG Russia strain and 60 control animals received sterile saline. Blood sampling was performed at time 0, previous to vaccination, and then at 3, 6, 9, 12, 15 and 18 months post-inoculation. The follow up of the IFN-ℽ response in animals determined that the BCG vaccination interferes with the diagnosis of bTB using the traditional bovine PPD between 9 and 12 months post-inoculation. Furthermore, the sensitization with non-tuberculous mycobacteria (NTM) was also interfering the diagnosis relying in PPDs, suggesting the need of using DIVA antigens under this epidemiological condition, whether or not the BCG vaccine is administered in cattle, in order to improve the accuracy of bTB diagnosis in central Chile.

Abstract The inspection of products of animal origin, carried out by veterinarian agricultural inspectors, is based on the Brazilian legislation described in the Brazilian Regulation for the Industrial and Sanitary Inspection of Animal Origin Products (RIISPOA) of the Ministry of Agriculture, Livestock, and Food Supply (MAPA). Decree No. 9,013, which revokes Decree No. 30,691 of 1952, was published on March 29, 2017, and major changes occurred in the legislation described in RIISPOA, such as the criteria for judging carcasses. This study aimed to verify whether these changes had an impact on the condemnations of bovine carcasses due to tuberculosis and cysticercosis in the State of Rio Grande do Sul, RS, Brazil. The number of slaughtered animals and the number of carcasses condemned due to tuberculosis and cysticercosis from 2014 to 2020 were obtained from the Agricultural Defense System (SDA) database, the official data system of the Secretariat of Agriculture, Livestock, and Rural Development (SEAPDR). An increase in the number of cases of carcasses condemned due to cysticercosis and a reduction of animals affected by the disease were observed in the period after changes in the regulation. The number of carcasses condemned due to tuberculosis and the number of animals affected by the disease remained stable after changes in the regulation. The results showed that changes in federal legislation had a real impact on the historical series of condemnations of bovine carcasses due to cysticercosis in the State of Rio Grande do Sul, while the impact of condemnations on carcasses affected by tuberculosis was not significant. origin inspectors RIISPOA (RIISPOA Agriculture Livestock MAPA. MAPA . (MAPA) No 9013 9 013 9,013 30691 30 691 30,69 1952 29 2017 Sul RS Brazil 201 202 SDA (SDA database SEAPDR. SEAPDR (SEAPDR) regulation significant (MAPA 901 01 9,01 3069 3 69 30,6 195 2 20 (SEAPDR 90 0 9,0 306 6 30, 19 9, 1

Resumo A inspeção de produtos de origem animal, realizada por médicos veterinários fiscais agropecuários, é baseada na legislação brasileira descrita no Regulamento da Inspeção Industrial e Sanitária de Produtos de Origem Animal (RIISPOA) do Ministério da Agricultura, Pecuária e Abastecimento (MAPA). No dia 29 de março de 2017 foi publicado o Decreto nº 9.013, que revoga o Decreto nº 30.691, de 1952, com isso grandes alterações ocorreram na legislação descrita no RIISPOA, como os critérios de julgamento de carcaças. O objetivo desse estudo foi verificar, se essas alterações tiveram impacto nas condenações de carcaças de bovídeos por cisticercose e tuberculose no Estado do Rio Grande do Sul (RS). O número de animais abatidos e o número de carcaças condenadas por tuberculose e cisticercose entre 2014 e 2020 foram obtidos do banco de dados do Sistema de Defesa Agropecuária (SDA), sistema oficial de dados da Secretaria da Agricultura Pecuária e Desenvolvimento Rural (SEAPDR), foi possível observar aumento do número de casos de carcaças condenadas por cisticercose e redução de animais afetados pela enfermidade no período após as alterações no regulamento. O número de carcaças condenadas por tuberculose, assim como o número de animais afetados pela doença se manteve estável após as mudanças no regulamento. Os resultados encontrados mostraram que as modificações na legislação federal trouxeram impacto real na série histórica de condenações de carcaças bovídeas por cisticercose no âmbito do Estado do Rio Grande do Sul enquanto nas carcaças afetadas por tuberculose o impacto das condenações não foi significativo. animal agropecuários RIISPOA (RIISPOA MAPA. MAPA . (MAPA) 2 201 9013 9 013 9.013 30691 30 691 30.691 1952 verificar RS. RS (RS) 202 SDA, SDA , (SDA) SEAPDR, SEAPDR (SEAPDR) regulamento significativo (MAPA 20 901 01 9.01 3069 3 69 30.69 195 (RS (SDA (SEAPDR 90 0 9.0 306 6 30.6 19 9. 30. 1

Abstract Apoptosis is a normally occurring process characterized by morphological changes in the cell. The species of the Mycobacterium tuberculosis complex induce apoptosis. In bovines, Mycobacterium bovis and its protein extracts induce apoptosis independently of caspase activation. However, the identity of the proteins in the extracts is unknown. In this study, we used size-exclusion chromatography to separate protein fractions from the culture filtrate. We obtained protein fractions that induced DNA fragmentation in bovine macrophages independently of caspase-3 activation. Liquid chromatography-mass spectrometry (LC-MS) identified three candidates responsible for the biological activity: MPB70, MPB83, and 60 kDa chaperonin.

Resumen La apoptosis es un proceso que ocurre de manera normal en los tejidos. Se caracteriza por cambios morfológicos en la célula. Las especies del complejo Mycobacterium tuberculosis inducen este fenómeno; en el modelo bovino, se demostró que tanto Mycobacterium bovis como sus extractos proteicos, inducen apoptosis independientemente de la activación de las caspasas, sin embargo, la identidad de las proteínas en los extractos proteicos se desconoce. En este estudio reportamos que, a través de cromatografía de exclusión por peso molecular, se obtuvieron fracciones proteicas con la capacidad de inducir fragmentación del DNA en macrófagos de bovinos independientemente de la activación de la caspasa 3. Asimismo, mediante cromatografía líquida-espectrometría de masas (LC-MS) se identificó la presencia de tres candidatos responsables de la actividad biológica: MPB70, MPB83 y chaperonina de 60 kDa.

RESUMO O objetivo do trabalho foi pesquisar Mycobacterium bovis em lesões sugestivas de tuberculose nas carcaças de bovinos no estado do Ceará, por meio dos testes de diagnóstico bacteriológico e molecular. Entre agosto de 2017 e janeiro de 2019, o Serviço de Inspeção Estadual (SIE) inspecionou 59.512 bovinos; destes, 7,4% (44/59.512) apesentaram lesões sugestivas. Desses animais foram enviadas 68 amostras, das quais 4,5% (31/68) estavam localizadas no pulmão, 2,9% (20/68) nos linfonodos, 2,0% (14/68) no fígado e 0,4% (3/68) na carcaça. Ao realizar o isolamento bacteriológico, 15,9% (7/44) dos bovinos evidenciaram crescimento de colônias nas amostras. Os esfregaços dos isolados foram submetidos à coloração de Zielh-Neelsen e todos eles confirmaram bacilo álcool-ácido resistente. A reação em cadeia da polimerase identificou todos os isolados, 100% (7/7), como M. bovis. A associação das técnicas de diagnóstico permitiu identificar a presença do agente no estado, e a análise molecular demonstrou ser uma técnica benéfica no monitoramento da tuberculose bovina, podendo ser utilizada como um método auxiliar no programa de controle e erradicação da tuberculose bovina no estado do Ceará.

ABSTRACT The objective of this research was to identify Mycobacterium bovis in lesions suggestive of tuberculosis in bovine carcasses in the State of Ceará, by means of bacteriological and molecular diagnostic tests. Between August 2017 and January 2019, the State inspection service (SIE) inspected 59,512 cattle, of which 7.4% (44 / 59,512) presented suggestive lesions. Of these animals, 68 samples were sent, of which 4.5% (31/68) located in the lung, 2.9% (20/68) in lymph nodes, 2.0% (14/68) in the liver, and 0.4% in the carcass (3/68). When performing bacteriological isolation, 15.9% (7/44) of bovines showed colony growth in the samples. The smears of the isolates were submitted to Zielh-Neelsen staining and all confirmed acid-fast bacilli. The polymerase chain reaction identified all isolates 100% (7/7) as M. bovis. The association of diagnostic techniques allowed to identify the presence of the agent in the State and the molecular analysis proved to be a beneficial technique in the monitoring of bovine tuberculosis and can be used as an auxiliary method in the bovine tuberculosis control and eradication program in the State of Ceará.

RESUMO: A tuberculose bovina é um problema econômico mundial e de saúde, que requer métodos de diagnóstico precisos para controle e erradicação. Objetivou-se com este estudo avaliar o desempenho de um teste comercial de ELISA no diagnóstico da tuberculose bovina. Foram avaliados pelo teste cervical comparativo (TCC) 1644 bovinos, provenientes de oito rebanhos de exploração leiteira, sendo três deles sem histórico recente da doença, e outros cinco com resultados positivos no último exame de tuberculinização. Para o diagnóstico sorológico da tuberculose utilizou-se um kit comercial de ELISA Mycobacterium bovis antibody test (ELISA M. bovis). Amostras de soro sanguíneo de 846 bovinos provenientes dos mesmos rebanhos foram também avaliadas no ELISA M. bovis. Os bovinos positivos no TCC e/ou ELISA M. bovis foram encaminhados para o abate sanitário. Dos positivos no ELISA para M. bovis foi realizado o exame macroscópico das carcaças e classificação das mesmas em: com presença ou ausência de lesões. Amostras de pulmão, fígado, e linfonodos, foram colhidas em duplicata para realização dos exames de cultivo microbiológico com posterior confirmação por PCR, e exame histopatológico com coloração de hematoxilina e eosina (HE). Dos 1,644 bovinos, 61 foram considerados positivos e 65 inconclusivos no TCC. O reteste dos inconclusivos identificou mais 19 positivos. No total, 80 (4,8%) bovinos positivos no TCC, provenientes de 5 rebanhos foram encaminhados para abate. O ELISA para M. bovis identificou 4,2% (36/846) de bovinos positivos, sendo 36 considerados negativos e um inconclusivo no exame de tuberculinização. Dos positivos no ELISA para M. bovis, 27 foram eutanasiados, e no exame macroscópico das carcaças 11% (3/27) dos animais apresentaram lesões sugestivas de tuberculose, e em apenas dois houve confirmação da doença pelos métodos histológico, microbiológico e PCR. A baixa associação dos resultados obtidos no ELISA para M. bovis com os exames macroscópico, histológico e isolamento microbiológico apontam para a fragilidade do desempenho do ELISA para o diagnóstico de tuberculose. Sugere-se assim que seu uso como método complementar para saneamento de rebanhos, seja adotado com cautela e considere a situação epidemiológica local.

ABSTRACT: Bovine tuberculosis is an economic and health problem, requiring precise diagnostic methods for its control and eradication. The aim of this study was to evaluate the performance of a commercial enzyme-linked immunosorbent assay (ELISA) test for the diagnosis of bovine tuberculosis. A total of 1,644 cattle from eight dairy herds were evaluated using the comparative cervical tuberculin test (CCTT). Three of the herds had no recent tuberculosis infection, and the other five had shown positive results in a previous tuberculin test. For the serological diagnosis of tuberculosis, a commercial ELISA antibody test kit for Mycobacterium bovis was used. Serum samples from 846 cattle from the eight herds were evaluated using ELISA for M. bovis. Animals that were positive based on either CCTT or ELISA for M. bovis or both were sent to slaughter. Samples of their lungs, livers, and lymph nodes were collected and stored under refrigeration for microbiological culture and subsequent confirmation by polymerase chain reaction. Samples from the same tissues were also fixed with 10% formaldehyde in bottles for histopathological examination and stained with hematoxylin and eosin (HE). Of the 1,644 cattle, 61 were considered positive and 65 inconclusive based on CCTT. Retesting of the inconclusive samples identified an additional 19 positive cases, totaling 80 (4.8%) CCTT-positive animals from five herds. ELISA for M. bovis identified 4.2% (36/846) positive cattle, of which 35 were considered negative and one inconclusive based on CCTT. Of the 36 positive cases identified by ELISA for M. bovis, 27 were euthanized, 11% (3/27) showed suggestive lesions of tuberculosis on macroscopic examination, and two were confirmed by histological, microbiological, and PCR methods. The weak association of ELISA for M. bovis with the results obtained by macroscopic, histological, and microbiological isolation indicates the fragility of ELISA performance in field conditions. Therefore, it is suggested that its use as a complementary method for herd sanitation be based on the local epidemiological situation.

Resumen Antecedentes: La tuberculosis bovina (TBB) y la brucelosis están asociadas con problemas persistentes en la ganadería Colombiana e incluso en los países desarrollados. La vigilancia de enfermedades en tiempo real es una estrategia clave para controlar y erradicar brotes de enfermedades infecciosas. Objetivo: Diseñar una herramienta epidemiológica para monitorear TBB y brucelosis en Colombia. Métodos: Se diseñó un panel de control interactivo para el mapeo de ambas enfermedades durante el periodo de observación entre los años 2004 y 2019. Resultados: El análisis de la herramienta mostró que las Provincias de Cundinamarca y Valle del Cauca han sido áreas epidémicas para TBB y brucelosis, respectivamente (p<0,001). Además, se encontró un aumento de la detección de casos de TBB en 2012 y de brucelosis durante 2019 (p<0,001). Conclusiones: Este panel epidemiológico permite el seguimiento de puntos críticos de TBB y tuberculosis, identificando sus tendencias a lo largo del tiempo, y proporciona información útil para las autoridades de sanidad animal que diseñan nuevas estrategias para los programas de control.

Resumo Antecedentes: A tuberculose bovina (TBB) e a brucelose estão associadas a problemas persistentes no campo da pecuária na Colômbia e até em países desenvolvidos. Portanto, a vigilância de doenças em tempo real é uma estratégia essencial para controlar e erradicar surtos de doenças infecciosas. Objetivo: Projetar uma ferramenta epidemiológica para monitorar a TB e a brucelose na Colômbia. Métodos: Um painel de controle interativo foi projetado para o mapeamento de ambas as doenças entre 2004 e 2019 como período de observação. Resultados: A análise da ferramenta mostrou que as Províncias de Cundinamarca e Valle del Cauca foram áreas epidêmicas para TBB e brucelose, respectivamente (p<0,001). Além disso, foi encontrado um aumento na detecção de casos em 2012 para TBB e brucelose durante 2019 (p<0,001). Conclusões: Esse painel epidemiológico poderia permitir o monitoramento de pontos críticos dessas doenças, identificando tendências ao longo do tempo, fornecendo informações úteis para as autoridades de saúde animal que elaboram novas estratégias para programas de controle.

Abstract Background: Bovine tuberculosis (BTB) and brucellosis are associated with devastating losses in the livestock sector in Colombia and even in developed countries. Real-time disease surveillance is a key strategy to control and eradicate infectious disease outbreaks. Objective: To design an epidemiological tool for monitoring BTB and brucellosis in Colombia. Methods: An interactive platform for disease mapping of BTB and brucellosis during an observation period between years 2004 and 2019 was designed. Results: Our analysis showed that the provinces of Cundinamarca and Valle del Cauca are regions affected by BTB and brucellosis epidemics, respectively (p<0.001). Furthermore, increased case detection of BTB was reported in 2012 and brucellosis in 2019 (p<0.001). Conclusions: This epidemiological platform allows tracking BTB and tuberculosis hotspots, identifying trends over time, and provides useful information to animal health authorities for designing new strategies in control programs.

Abstract Bovine tuberculosis is a disease caused by Mycobacterium bovis that affects cattle and other species, including humans. Mycobacterium bovis resides mainly in macrophages, so bacilli survival within macrophages is related to virulence. Isolation and strain identification are important for disease control. However, little is known about virulence of the circulating strains in cattle populations. Therefore, the aim of this study was to compare the intracellular survival of Mycobacterium bovis strains with high and low frequency genotypes in cattle in Mexico. Four high frequency genotypes and four low frequency genotypes were identified and subjected to intracellular survival assays in bovine macrophages. Results showed that the phagocytosis proportion was approximately 63 % for all strains. There were no significant differences in the average Colony Forming Units (CFUs) in phagocytosis and survival between the high and low frequency groups; however, when the CFU average of phagocytosis was compared with the survival, significant differences were found in both groups. In intracellular growth, a significant difference was observed between low and high frequency strains, and between low frequency strains. Finally, the intracellular growth average of the groups was analyzed showing no significant difference. These results suggest that the frequency of the genotype in cattle population is not related to the intracellular survival and the virulence of the M. bovis strains.

Resumen Causada por Mycobacterium bovis, la tuberculosis bovina es una enfermedad que afecta al ganado y a otras especies, incluidos los humanos. M. bovis reside principalmente en macrófagos, por lo que la supervivencia de los bacilos dentro de los macrófagos está relacionada con la virulencia. El aislamiento y la identificación de cepas son importantes para el control de las enfermedades causadas por M. bovis. Se sabe poco sobre la virulencia de las cepas de M. bovis circulantes en las poblaciones de ganado. En este estudio se comparó la supervivencia intracelular de cepas de M. bovis de genotipos que se encuentran en altas y bajas frecuencias en los bovinos en México. Se identificaron cuatro genotipos de alta frecuencia y cuatro de baja frecuencia a las cuales se sometieron a ensayos de supervivencia intracelular en macrófagos bovinos. La proporción de fagocitosis fue de aproximadamente 63% para todas las cepas. No hubo diferencias entre los grupos de alta y baja frecuencia en cuanto al promedio de las unidades formadoras de colonias (UFC) en fagocitosis y supervivencia. Sin embargo, el promedio de UFC de fagocitosis sí difirió de UFC de supervivencia en ambos grupos. El crecimiento intracelular fue diferente entre las cepas de baja y alta frecuencia, pero también entre las cepas de baja frecuencia. El promedio de crecimiento intracelular no fue diferente entre las cepas de alta y baja frecuencia. Estos resultados sugieren que la supervivencia intracelular y la virulencia de las cepas de M. bovis evaluadas no están relacionadas con la frecuencia de los genotipos en una población bovina.

ABSTRACT A clinical case of tuberculous meningoencephalitis in a 5-month-old Holstein calf that presented nervous symptoms is described. The calf was part of a herd with a 30% prevalence of tuberculosis. The animal was euthanized, and a necropsy was performed. No lesions were found in organs of the thoracic or abdominal cavity. Brain samples were taken for histopathology (hematoxylin-eosin and Zielh Nelsen staining), bacteriology and PCR-MRD1 studies. In the brain, macroscopic inflammatory lesions with congestion and granular appearance were observed in the meninges, as well as fibrous adhesions in the ventral area. Microscopically, inflammatory infiltrate, lymphocytes, macrophages, epithelioid cells, plasma cells and multinucleated giant cells, areas of central necrosis, dystrophic mineralization and acid-fast bacilli were observed. Granulomatous meningoencephalitis due to Mycobacterium bovis was confirmed by bacterial isolation and PCR-MRD1.

RESUMEN Se describe un caso clínico de meningoencefalitis tuberculosa en un ternero Holstein de cinco meses de edad que presentaba signología nerviosa; además de pertenecer a un hato con 30% de prevalencia a tuberculosis. Se llevó a cabo la eutanasia del animal y se practicó la necropsia. No se encontraron lesiones en órganos de cavidad torácica ni abdominal. Se tomaron muestras de encéfalo para estudios de histopatología (tinción con hematoxilina-eosina y Zielh Nelsen), bacteriología y PCR-MRD1. En el encéfalo se observaron lesiones inflamatorias macroscópicas con congestión y aspecto granular en meninges, así como adherencias fibrosas en la zona ventral. Microscópicamente se observó infiltrado inflamatorio, linfocitos, macrófagos, células epitelioides, células plasmáticas y células gigantes multinucleadas, áreas de necrosis central, mineralización distrófica y bacilos ácido-alcohol resistentes. La meningoencefalitis granulomatosa por Mycobacterium bovis se confirmó por aislamiento bacteriano y PCR-MRD1.

ABSTRACT: Bovine brucellosis is caused by Brucella abortus. The bacterial pathogen causes economic losses because it induces abortion in cattle. Vaccination of calves with live B. abortus strain 19 induces a certain level of protection but induces persistent antibodies against cell envelope lipopolysaccharide that make it difficult to Distinguish Infected from Vaccinated Animals (DIVA). Live vaccine B. abortus strain RB51 was developed to elimínate such interfering antibodies and therefore, facilitate the differentiation of infected from vaccinated animals and help in the eradication of the disease. Vaccination with strain RB51 induces levels of protection similar to strain 19 but neither of the two vaccines give complete protection. We have been working to enhance protection induced by strain RB51 vaccine. Protective Brucella antigens can be over-expressed in strain RB51 by introducing a plasmid containing the leuB gene and the genes encoding such antigens. To avoid the expression of antibiotic resistance genes, we produced a leuB deficient strain RB51 and introduced a plasmid containing the leuB gene and the genes to be over-expressed. This new strain maintains the plasmid and has induced significantly high protection levels in mice. In addition, it allowed the construction of an RB51 vaccine strain able to express Mycobacterium bovis protective antigens so that the vaccine could protect against brucellosis and tuberculosis simultaneously.

ABSTRACT The diagnosis of bovine tuberculosis (TB) by molecular techniques has been broadly studied. These methods allow accelerating the diagnosis, in addition to presenting high specificity and sensitivity in the identification of the pathogen, critical characteristic for public health, especially when it comes to the direct diagnosis of the biologic samples, which has been little explored. This paper has evaluated a multiplex polymerase chain reaction (mPCR) as a tool to diagnose TB, which was performed directly on the granulomatous material of suspicious lesions collected in a cold chamber under state inspection in the state of Bahia, Brazil. Of the 74 samples evaluated, 14.86% were positive, with 10.81% positive for mPCR and culture, 4.05% negative for cultivation and positive for mPCR. The correlation between the cultivation and the mPCR presented agreeance higher than 61.54% of the cases. The results have indicated that the protocol proved itself effective, fast and very promising in the surveillance in slaughterhouses for the diagnosis of tuberculosis directly from the granuloma.

RESUMO: A aplicação de análises espacial e temporal em epidemiologia visa compreender as causas e consequências da heterogeneidade espacial e temporal nos estudos de doenças infecciosas. A tuberculose bovina (TBb) é uma doença infecciosa crônica e progressiva causada pelo Mycobacterium bovis e uma importante zoonose em todo o mundo. O objetivo deste estudo foi realizar análises espaciais e temporais em um banco de dados secundário de casos positivos de TBb registrados pelo Serviço de Inspeção Federal em dois frigoríficos da mesorregião Oeste de Minas, estado de Minas Gerais, Brasil, de 2008 a 2012 e sugerir sua utilização na vigilância epidemiológica. Os bovinos abatidos com lesões macroscópicas de TBb durante a inspeção post mortem foram considerados positivos neste estudo. Os dados utilizados foram os casos positivos cadastrados no Sistema de Informações Gerenciais do Serviço de Inspeção Federal do Ministério da Agricultura, Pecuária e Abastecimento (SIGSIF/MAPA), o número de animais abatidos mensalmente e seus municípios de origem, por frigorífico. Foi calculada a prevalência (%) de casos por bovinos abatidos e avaliada a relação entre esses casos de TBb e os casos de tuberculose humana (TBh) e o PIB per capita das mesorregiões que compõem os municípios pesquisados. A prevalência foi de 1.030 casos de TBb (0,28%). Os menores números de casos e prevalências foram obtidos em 2010, enquanto as maiores prevalências foram observadas em 2011. O Frigorífico 1 apresentou estacionariedade, enquanto o Frigorífico 2 apresentou tendência anual crescente para os dados desde outubro de 2009. Correlação estatística entre o número de casos de TBb e TBh (p=0,006, r=0,148) foi observada. As prevalências nos Frigoríficos 1 e 2 não se correlacionaram (p>0,05). Os municípios de origem demonstraram aleatoriedade em sua distribuição espacial. O SIGSIF/MAPA pode ser utilizado para a análise da distribuição espacial e temporal a fim de contribuir com o monitoramento dos serviços de saúde animal com informações sobre a prevalência de condenações por TBb nas mesorregiões Oeste de Minas, Sul/Sudoeste de Minas, Campo das Vertentes e Zona da Mata de Minas Gerais, Brasil.

ABSTRACT: The application of spatial and temporal analysis in epidemiology aims to understand the causes and consequences of spatial and temporal heterogeneity in studies of infectious diseases. Bovine tuberculosis (bTB) is a chronic and progressive infectious disease caused by Mycobacterium bovis and is an important zoonosis worldwide. The aim of this study was to conduct spatial and temporal analyses of a secondary database of bTB-positive cases registered by the Federal Inspection Service in two slaughterhouses from the West of Minas mesoregion of Minas Gerais state, Brazil, from 2008 to 2012 and to suggest its use in epidemiological surveillance. The culled cattle with bTB macroscopic lesions during post mortem inspection were considered positive in this study. The data used were the positive cases registered on the “Sistema de Informações Gerenciais do Serviço de Inspeção Federal” (SIG/SIF - Management Information System of the Federal Inspection Service) of the “Ministério da Agricultura, Pecuária e Abastecimento” (MAPA - Ministry of Agriculture, Livestock and Supplies), the number of animals slaughtered monthly and their municipalities of origin, per slaughterhouse. The prevalence (%) of cases per cattle slaughtered was calculated, and the relationship between these cases of bTB and cases of human tuberculosis (hTB) and the GDP per capita of the mesoregions comprising the municipalities surveyed was evaluated. The prevalence was 1,030 cases of bTB (0.28%). The lowest case numbers and prevalence rates were obtained in 2010, while the highest rates were observed in 2011. Slaughterhouse 1 showed stationarity, while Slaughterhouse 2 showed an increasing annual trend for data since October 2009. A statistical correlation between the numbers of cases of bovine and human TB (p=0.006, r=0.148) was observed. The prevalence for Slaughterhouses 1 and 2 were not correlated (p>0.05). The municipalities of origin demonstrated randomness in their spatial distribution. SIG/SIF-MAPA may be used for the analysis of spatial and temporal distribution to contribute to the monitoring of animal health services with information on the prevalence of culling for bTB in the West of Minas, South/Southwest of Minas, Campo das Vertentes and Zona da Mata mesoregions of Minas Gerais, Brazil.