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: 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: 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 (

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 diagnóstico rápido da tuberculose bovina (TB), permite a atuação de instituições oficiais de vigilância em áreas que possam constituir possíveis focos de propagação da doença. Essa pesquisa identificou o agente Mycobacterium bovis através do PCR multiplex das colônias isoladas. Através do serviço de inspeção oficial (Federal e Estadual), foram coletadas amostras sugestivas de TB durante a inspeção post-mortem de carcaças bovinas em matadouros-frigoríficos e estas foram encaminhadas ao laboratório para os diagnósticos complementares. As amostras foram comprovadas por meio de cultura bacteriológica e coloração Ziehl-Neelsen. Nos isolados caracterizados por Bacilos ÁcidoÁlcool Resistente (BAAR), aplicou-se a técnica de PCR multiplex visando a confirmação do agente Mycobacterium bovis. Foram analisadas 991.316 carcaças de bovinos. Entre estas, 32 apresentaram amostras sugestivas de TB que foram coletadas, provenientes de animais de 30 municípios. Entre as amostras coletadas, 28,1% (9/32) apresentaram crescimento em meios de cultura bacteriológicos com BAAR. Os BAAR isolados submetidos à PCR multiplex confirmaram a presença de M. bovis. A técnica de PCR multiplex associada ao exame bacteriológico e aplicada aos achados post-mortem permitiu o diagnóstico de M. bovis nos municípios amostrados do estado da Bahia. TB, , (TB) doença isoladas Federal Estadual, Estadual Estadual) postmortem post mortem matadourosfrigoríficos matadouros frigoríficos complementares ZiehlNeelsen. ZiehlNeelsen Ziehl Neelsen. Neelsen Ziehl-Neelsen BAAR, (BAAR) aplicouse aplicou se 991316 991 316 991.31 bovinos 3 281 28 1 28,1 9/32 932 9 (9/32 M Bahia (TB (BAAR 99131 99 31 991.3 2 28, 9/3 93 (9/3 9913 991. 9/ (9/ (9 (

Abstract The rapid diagnosis of bovine tuberculosis (BT) allows official surveillance institutions to act in areas that could be potential hotspots for the spread of the disease. The study aimed to identify the Mycobacterium bovis agent by multiplex PCR of isolated colonies. Through the official inspection service (Federal/State), samples suggestive of BT were collected during the post-mortem inspection of bovine carcasses in slaughterhouses and sent to the laboratory for complementary diagnostics. The samples were analyzed using bacteriological culture and Ziehl-Neelsen staining. In isolates characterized as Acid-Alcohol Resistant Bacilli (BAAR), the multiplex PCR technique confirmed the agent Mycobacterium bovis. Nine hundred ninety-one thousand three hundred sixteen cattle carcasses were inspected, collecting 32 BT-suggestive samples in from animalsfrom 30 municipalities. Of these, 28.1% (9/32) showed growth on bacteriological culture media with BAAR. The BAAR isolates submitted to multiplex PCR confirmed the presence of M. bovis. The multiplex PCR technique associated with bacteriological examination and applied to post-mortem findings enabled the diagnosis of M. bovis in the municipalities sampled in the Bahia state. (BT disease colonies Federal/State, FederalState Federal/State , Federal State (Federal/State) postmortem post mortem diagnostics ZiehlNeelsen Ziehl Neelsen staining AcidAlcohol Acid Alcohol BAAR, (BAAR) ninetyone ninety one inspected 3 BTsuggestive these 281 28 1 28.1 9/32 932 9 (9/32 M state (Federal/State (BAAR 2 28. 9/3 93 (9/3 9/ (9/ (9 (

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.

Resumen La vacuna BCG es clave para el control de la tuberculosis. En ocasiones se observan eventos adversos sistémicos causados por Mycobacterium bovis BCG; usualmente asociados a inmunodeficiencia. Describimos seis casos clínicos de niños vacunados con BCG al nacer, con complicaciones sistémicas post-vacunación. Método: Revisión de historias clínicas de pacientes con infección por M. bovis BCG atendidos en un hospital pediátrico, entre 2010 y 2019. Resultados: De 400 casos confirmados de infecciones por complejo Mycobacterium tuberculosis; ocho fueron identificados como M. bovis BCG; seis casos correspondieron a eventos adversos sistémicos post-vacuna BCG: dos con lesiones cutáneas a distancia, dos osteomielitis y dos infecciones generalizadas. En cinco de los seis pacientes se detectó una alteración de la respuesta inmune. Un paciente falleció por falla multiorgánica, uno se derivó y cuatro completaron 12 meses de tratamiento: dos meses de isoniacida, rifampicina, etambutol, y moxifloxacina, y 10 meses de isoniacida y rifampicina. Tuvieron buena tolerancia a los medicamentos, sin recaída a los dos años. Conclusión: La infección grave por M. bovis BCG es una rara complicación sistémica de la vacunación. Es razonable buscar defectos inmunológicos en los niños que desarrollan este tipo de eventos adversos.

Abstract The BCG vaccine is key to tuberculosis control. Systemic adverse events caused by Mycobacterium bovis BCG are occasionally observed; usually associated with immunodeficiency. In this report we describe six cases of children vaccinated with BCG at birth, with post-vaccination systemic complications. Method: retrospective review of medical records of patients with M. bovis BCG infection treated in a pediatric hospital between 2010 and 2019. Results: Of 400 confirmed cases of Mycobacterium tuberculosis complex infection, eight identified as M. bovis BCG, six corresponded to systemic adverse events post-BCG vaccine: two distant skin lesions, two osteomyelitis and two generalized infections. An altered immune response was detected in five of the six patients. One patient died of multiorgan failure, one was referred and four completed 12 months of treatment: two months of isoniazid, rifampin, ethambutol, and moxifloxacin, and 10 months of isoniazid and rifampin. They had good tolerance to medications, without relapse at two years. Conclusion Serious M. bovis BCG infection is a rare systemic complication of vaccination. It is reasonable to look for immunological defects in children who develop these types of adverse events.

BACKGROUND The Bacille Calmette-Guérin (BCG) vaccine comprises a family of strains with variable protective efficacy against pulmonary tuberculosis (TB) and leprosy, partly due to genetic differences between strains. OBJECTIVES Previous data highlighting differences between the genomes and proteomic profiles of BCG strains Moreau and Pasteur led us to evaluate their behaviour in the macrophage microenvironment, capable of stimulating molecular responses that can impact the protective effect of the vaccine. METHODS Strain infectivity, viability, co-localisation with acidified vesicles, macrophage secretion of IL-1 and MCP-1 and lipid droplet biogenesis were evaluated after infection. FINDINGS We found that BCG Moreau is internalised more efficiently, with significantly better intracellular survival up to 96 h p.i., whereas more BCG Pasteur bacilli were found co-localised in acidified vesicles up to 6 h p.i. IL-1β and MCP-1 secretion and lipid droplet biogenesis by infected macrophages were more prominent in response to BCG Pasteur. MAIN CONCLUSION Overall, our results show that, compared to Pasteur, BCG Moreau has increased fitness and better endurance in the harsh intracellular environment, also regulating anti-microbial responses (lower IL-1b and MCP-1). These findings contribute to the understanding of the physiology of BCG Moreau and Pasteur in response to the intraphagosomal environment in a THP-1 macrophage model. CalmetteGuérin Calmette Guérin (BCG TB (TB leprosy microenvironment infectivity viability colocalisation co localisation IL1 IL 1 IL- MCP1 MCP MCP- infection efficiently 9 pi p i colocalised localised p.i IL1β ILβ 1β β Overall antimicrobial anti microbial lower IL1b ILb 1b b MCP1. . MCP-1) THP1 THP THP- model

RESUMO Objetivo: As crianças são um grupo demográfico importante para a compreensão da epidemiologia da tuberculose em geral, e o monitoramento da tuberculose infantil é essencial para a prevenção adequada. O presente estudo procurou caracterizar a distribuição espacial das taxas de notificação de tuberculose infantil em Portugal continental; identificar áreas de alto risco e avaliar a associação entre taxas de notificação de tuberculose infantil e privação socioeconômica. Métodos: Por meio de modelos espaciais hierárquicos bayesianos, analisamos a distribuição geográfica das taxas de notificação de tuberculose pediátrica em 278 municípios entre 2016 e 2020 e determinamos as áreas de alto e baixo risco. Usamos a versão portuguesa do European Deprivation Index para calcular a associação entre a tuberculose infantil e a privação socioeconômica em cada área. Resultados: As taxas de notificação variaram de 1,8 a 13,15 por 100.000 crianças com idade < 5 anos. Identificamos sete áreas de alto risco, cujo risco relativo era significativamente maior que a média da área de estudo. Todas as sete áreas de alto risco situavam-se na área metropolitana do Porto e de Lisboa. Houve uma relação significativa entre a privação socioeconômica e as taxas de notificação de tuberculose pediátrica (risco relativo = 1,16; intervalo de credibilidade de 95%: 1,05-1,29). Conclusões: Áreas identificadas como sendo de alto risco e desfavorecidas socioeconomicamente devem constituir áreas-alvo para o controle da tuberculose, e esses dados devem ser integrados a outros fatores de risco para definir critérios mais precisos para a vacinação com BCG. Objetivo geral adequada continental Métodos bayesianos 27 201 202 Resultados 18 1 8 1, 1315 13 15 13,1 100000 100 000 100.00 anos situavamse situavam se Lisboa 1,16 116 16 95% 95 1,051,29. 105129 1,05 1,29 . 05 29 1,05-1,29) Conclusões áreasalvo alvo BCG 2 20 131 13, 10000 10 00 100.0 1,1 11 9 051 1,051,29 10512 105 1,0 129 1,2 0 1,05-1,29 1000 100. 1,051,2 1051 12 1,05-1,2 1,051, 1,05-1, 1,051 1,05-1 1,05-

ABSTRACT Objective: Children are an important demographic group for understanding overall tuberculosis epidemiology, and monitoring of childhood tuberculosis is essential for appropriate prevention. The present study sought to characterize the spatial distribution of childhood tuberculosis notification rates in continental Portugal; identify high-risk areas; and evaluate the association between childhood tuberculosis notification rates and socioeconomic deprivation. Methods: Using hierarchical Bayesian spatial models, we analyzed the geographic distribution of pediatric tuberculosis notification rates across 278 municipalities between 2016 and 2020 and determined high-risk and low-risk areas. We used the Portuguese version of the European Deprivation Index to estimate the association between childhood tuberculosis and area-level socioeconomic deprivation. Results: Notification rates ranged from 1.8 to 13.15 per 100,000 children under 5 years of age. We identified seven high-risk areas, the relative risk of which was significantly above the study area average. All seven high-risk areas were located in the metropolitan area of Porto or Lisbon. There was a significant relationship between socioeconomic deprivation and pediatric tuberculosis notification rates (relative risk = 1.16; Bayesian credible interval, 1.05-1.29). Conclusions: Identified high-risk and socioeconomically deprived areas should constitute target areas for tuberculosis control, and these data should be integrated with other risk factors to define more precise criteria for BCG vaccination. Objective epidemiology prevention Portugal highrisk high Methods models 27 201 202 lowrisk low arealevel level Results 18 1 8 1. 1315 13 15 13.1 100000 100 000 100,00 age average Lisbon 1.16 116 16 interval 1.051.29. 105129 1.05 1.29 . 05 29 1.05-1.29) Conclusions control vaccination 2 20 131 13. 10000 10 00 100,0 1.1 11 051 1.051.29 10512 105 1.0 129 1.2 0 1.05-1.29 1000 100, 1.051.2 1051 12 1.05-1.2 1.051. 1.05-1. 1.051 1.05-1 1.05-

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 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.

Abstract The objective of this study was to evaluate the anti-bacterial and antimycobacterial potential of the by-products of white shrimp (Litopenaeus vannamei). The following extracts were obtained: exoskeleton hexanic, methanolic and acqueous extracts (ExHex, ExMe, ExAc); and cephalothorax hexanic, acetonic and methanolic extracts (CeHex, CeAce, CeMe). Antibacterial effect was determined by the broth microdilution method against Gram-positive bacteria: Enterococcus faecalis American Type Culture Collection (ATCC) 51299, Staphylococcus aureus ATCC 25293, and Staphylo-coccus epidermidis; Gram-negative bacteria: Escherichia coli ATCC 25922, Klebsiella pneumoniae, Pseudomonas aeruginosa ATCC 10145, and Salmonella typhimurium; and Mycobacterium bovis bacillus Calmette-Guérin (M. bovis BCG) Danish strain. CeHex resulted active against all Gram-positive and Gram-negative bacteria (MIC50= 400 µg mL-1) and against M. bovis BCG (MIC100= 250 µg mL-1). Gas chromatography (GC) of CeHex identified oleic, linoleic, palmitic, stearic, behenic, palmitoleic and linolenic fatty acids. The strong antibacterial activity of CeHex and the identification of its main chemical constituents justify further studies on the clinical applications of this marine by-product.

Resumen El objetivo de este estudio fue evaluar el potencial antibacteriano y antimicobacteriano de los sub-productos del camarón blanco (Litopenaeus vannamei). Fueron obtenidos los siguientes extractos: extracto hexánico, metanólico y acuoso del exoesqueleto (ExHex, ExMe, ExAc); y extracto hexánico, acetónico y metanólico del cefalotórax (CeHex, CeAce, CeMe). El efecto antibacteriano fue evaluado mediante el método de microdilución en caldo contra las bacterias Gram-positivas: Enterococcus faecalis American Type Culture Collection (ATCC) 51299, Staphylococcus aureus ATCC 25293, y Staphylococcus epidermidis; bacterias Gram-negativas: Escherichia coli ATCC 25922, Klebsiella pneumoniae, Pseudomonas aeruginosa ATCC 10145, y Salmonella typhimurium; y Mycobacterium bovis bacilo Calmette-Guérin (M. bovis BCG) cepa Danesa. CeHex resultó activo contra todas las bacterias Gram-positivas y Gram-negativas (MIC50= 400 µg mL-1) y contra M. bovis BCG (MIC100= 250 µg mL-1). Mediante cromatogra-fía de gases (GC) de CeHex se identificaron los ácidos grasos: oleico, linoleico, palmítico, esteárico, behénico, palmitoleico y linolénico. La fuerte actividad antibacteriana de CeHex y la identificación de sus principales components químicos justifican estudios posteriores en las aplicaciones clínicas de este sub-producto marino.

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.

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.