Ocular tuberculosis in a calf.

BACKGROUND: Bovine tuberculosis is a chronic inflammatory disease that causes granuloma formation mainly in retropharyngeal, tracheobronchial, mediastinal lymph nodes and lungs of bovines. The presence of these lesions in other tissues such as the eyeball is very rare and difficult to diagnose. This study describes macroscopic and microscopic pathological findings in a calf with ocular and meningeal tuberculosis. CASE PRESENTATION: March 2019, an eight-month-old Holstein Friesian calf was identified in a dairy farm located in central Mexico with a clinical cough, anorexia, incoordination, corneal opacity and vision loss. At necropsy, pneumonia, lymphadenitis, meningitis, and granulomatous iridocyclitis were observed. The histopathological examination revealed granulomatous lesions in lung tissue, lymph nodes, meninges and eyes with the presence of acid-fast bacilli associated with Mycobacterium spp. CONCLUSION: To the best of our knowledge, this is the first report that describes macroscopic and microscopic pathological findings of ocular tuberculosis in cattle. This report highlights the importance of considering bovine tuberculosis in the differential diagnosis of corneal opacity and loss of vision in cattle.

BoLA-DRB3 exon2 polymorphisms among tuberculous cattle: Nucleotide and functional variability and their association with bovine tuberculosis pathology.

Bovine tuberculosis (bTB) is caused by Mycobacterium bovis and disseminated worldwide. In Argentina, the highest prevalence occurs in dairy areas. BoLA DRB3.2 is related to the adaptive immunity in mycobacterial infections. Genetic polymorphisms of this marker have been associated with resistance or susceptibility to bovine diseases. We evaluated the association between BoLA DRB3.2 polymorphisms and bTB pathology scores in dairy and beef cattle breeds of Argentina. Most bovines exhibited visible lesions compatible with tuberculosis and, furthermore, 150 (85.7%) were also positive by bacteriology. A pathology index showed a variable degree of disease, from 3 to 76 (median pathology score = 9 (IQR: 7-15)). Thirty-five BoLA DRB3.2 alleles were identified with an associated frequency from 16% to 0.3%, distributed 73% (n = 128) in heterozygosis and 27% (n = 47) in homozygosis, with 12 BoLA DRB3.2 alleles (*0101, *1101, *1501, *0201, *2707 *1001, *1002, *1201, *14011, *0501 *0902 and *0701) representing the 74.7% of the population variability. A functional analysis grouped them in 4 out of 5 clusters (A-D), suggesting a functional overlapping. Among the 90 identified genotypes, *1101/*1101, *1101/*1501 and *0101/*0101 were the most frequent (10%, 8.9% and 8.9%, respectively). No association was detected between the pathology scores and a specific DRB3.2 allele (p > .05). Animals infected with M. bovis spoligotype SB0153 showed a significantly higher pathology score than those affected by the spoligotype SB0145 (p = .018). Furthermore, the Aberdeen Angus breed exhibited highest pathological scores (p < .0001), which were associated with disseminated lesion, thus suggesting that the host component could be important to the disease progression.

Epidemiological analysis of bovine tuberculosis in the southern region of Rio Grande do Sul from 2000 to 2015 / Análise epidemiológica da tuberculose bovina na região sul do Rio Grande do Sul no período de 2000 a 2015

Bovine tuberculosis (BTB) is an infectious and zoonotic disease with socioeconomic importance, responsible for economic losses in livestock due to the decrease in animal productivity, mortality, bovine carcass condemnation and restriction on exportations. The objective of the present study was to evaluate the epidemiological aspects of this disease through a descriptive analysis and time series of cases of bovine tuberculosis diagnosed at the Laboratório Regional de Diagnóstico (LRD) of the Faculdade de Medicina Veterinária, Universidade Federal de Pelotas (UFPel) from 2000 to 2015. A survey was carried out in the LRD database of cases diagnosed as tuberculosis in cattle during this period. The incidence rate of BTB ([cases/population] x 100000) was calculated. Qualitative variables (sex, age, race, origin, type of material and presumptive diagnosis) were analyzed in a descriptive way, aiming to establish the profile of affected animals, characterize the municipalities in which the disease is originated and also fix the main referral organs for diagnosis. The time series analysis was performed using the Gretl statistical software 1.9.12 (GNU Regression, Econometric and Time-series Library). To verify the existence of the stationary distribution, the test of Dickey-Fuller was used, considering a value of p<0.05. During this period, 331 cases of BTB were diagnosed in the LRD. The present study identified that the predominant profile of the bovines affected by tuberculosis was female bovine aged more than two years old and crossbreed; the main organ sent for histopathological examination was lymph nodes. The microregion of Pelotas presented the highest incidence of the disease. There was also a tendency of a decrease in the incidence rate of this disease over the years, absence of seasonal influence in the occurrence of the disease and no cases of epidemics occurred from 2002 to 2015 in the area covered by the LRD-UFPel.(AU)

Tuberculose bovina é uma enfermidade infectocontagiosa e zoonótica, de importância socioeconômica, responsável por prejuízos econômicos na pecuária em decorrência da diminuição na produtividade animal, mortalidades, condenações de carcaças e restrições nas exportações. O objetivo do presente estudo foi avaliar os aspectos epidemiológicos, por meio de uma análise descritiva e de séries temporais, dos casos de tuberculose bovina diagnosticados no Laboratório Regional de Diagnóstico da Faculdade de Veterinária, Universidade Federal de Pelotas (LRD-UFPel), no período de 2000 a 2015. Foi realizado um levantamento no banco de dados do LRD dos casos diagnosticados como tuberculose em bovinos, entre os anos de 2000 e 2015. Foi calculada a taxa de incidência da tuberculose bovina ([casos/população] x 100000). As variáveis qualitativas (sexo, idade, raça, procedência, tipo de material e diagnóstico presuntivo) foram analisadas de forma descritiva, tendo como finalidade estabelecer o perfil dos animais acometidos, caracterizar os municípios de origem, e estabelecer os principais órgãos remetidos para diagnóstico. A análise de série temporal foi feita através do software estatístico Gretl 1.9.12 (GNU Regression, Econometricand Time-series Library) e para verificar a existência da estacionariedade utilizou-se o teste de Dickey-Fuller aumentado, considerando um valor de p<0,05. Durante este período foram diagnosticados 331 casos de tuberculose bovina no LRD. O presente estudo permitiu concluir que o perfil predominante dos bovinos acometidos por tuberculose foi de fêmeas, com mais de dois anos de idade e mestiças; o principal órgão encaminhado para exame histopatológico foram linfonodos e a microrregião de Pelotas apresentou a maior incidência da doença. Foi constatada, ainda, uma tendência de queda na taxa de incidência da doença ao longo dos anos analisados, ausência de influência sazonal na ocorrência da enfermidade e, não foram registrados casos de epidemias nos anos de 2002 a 2015, na área de abrangência do LRD-UFPel.(AU)

Epidemiological analysis of bovine tuberculosis in the southern region of Rio Grande do Sul from 2000 to 2015 / Análise epidemiológica da tuberculose bovina na região sul do Rio Grande do Sul no período de 2000 a 2015

Bovine tuberculosis (BTB) is an infectious and zoonotic disease with socioeconomic importance, responsible for economic losses in livestock due to the decrease in animal productivity, mortality, bovine carcass condemnation and restriction on exportations. The objective of the present study was to evaluate the epidemiological aspects of this disease through a descriptive analysis and time series of cases of bovine tuberculosis diagnosed at the Laboratório Regional de Diagnóstico (LRD) of the Faculdade de Medicina Veterinária, Universidade Federal de Pelotas (UFPel) from 2000 to 2015. A survey was carried out in the LRD database of cases diagnosed as tuberculosis in cattle during this period. The incidence rate of BTB ([cases/population] x 100000) was calculated. Qualitative variables (sex, age, race, origin, type of material and presumptive diagnosis) were analyzed in a descriptive way, aiming to establish the profile of affected animals, characterize the municipalities in which the disease is originated and also fix the main referral organs for diagnosis. The time series analysis was performed using the Gretl statistical software 1.9.12 (GNU Regression, Econometric and Time-series Library). To verify the existence of the stationary distribution, the test of Dickey-Fuller was used, considering a value of p<0.05. During this period, 331 cases of BTB were diagnosed in the LRD. The present study identified that the predominant profile of the bovines affected by tuberculosis was female bovine aged more than two years old and crossbreed; the main organ sent for histopathological examination was lymph nodes. The microregion of Pelotas presented the highest incidence of the disease. There was also a tendency of a decrease in the incidence rate of this disease over the years, absence of seasonal influence in the occurrence of the disease and no cases of epidemics occurred from 2002 to 2015 in the area covered by the LRD-UFPel.(AU)

Tuberculose bovina é uma enfermidade infectocontagiosa e zoonótica, de importância socioeconômica, responsável por prejuízos econômicos na pecuária em decorrência da diminuição na produtividade animal, mortalidades, condenações de carcaças e restrições nas exportações. O objetivo do presente estudo foi avaliar os aspectos epidemiológicos, por meio de uma análise descritiva e de séries temporais, dos casos de tuberculose bovina diagnosticados no Laboratório Regional de Diagnóstico da Faculdade de Veterinária, Universidade Federal de Pelotas (LRD-UFPel), no período de 2000 a 2015. Foi realizado um levantamento no banco de dados do LRD dos casos diagnosticados como tuberculose em bovinos, entre os anos de 2000 e 2015. Foi calculada a taxa de incidência da tuberculose bovina ([casos/população] x 100000). As variáveis qualitativas (sexo, idade, raça, procedência, tipo de material e diagnóstico presuntivo) foram analisadas de forma descritiva, tendo como finalidade estabelecer o perfil dos animais acometidos, caracterizar os municípios de origem, e estabelecer os principais órgãos remetidos para diagnóstico. A análise de série temporal foi feita através do software estatístico Gretl 1.9.12 (GNU Regression, Econometricand Time-series Library) e para verificar a existência da estacionariedade utilizou-se o teste de Dickey-Fuller aumentado, considerando um valor de p<0,05. Durante este período foram diagnosticados 331 casos de tuberculose bovina no LRD. O presente estudo permitiu concluir que o perfil predominante dos bovinos acometidos por tuberculose foi de fêmeas, com mais de dois anos de idade e mestiças; o principal órgão encaminhado para exame histopatológico foram linfonodos e a microrregião de Pelotas apresentou a maior incidência da doença. Foi constatada, ainda, uma tendência de queda na taxa de incidência da doença ao longo dos anos analisados, ausência de influência sazonal na ocorrência da enfermidade e, não foram registrados casos de epidemias nos anos de 2002 a 2015, na área de abrangência do LRD-UFPel.(AU)

Bovine tuberculosis: diagnosis in dairy cattle through the association of analyzes / Tuberculose bovina: diagnóstico em bovinos leiteiros através da associação de análises

Tuberculosis is a chronic anthropozoonosis of worldwide occurrence, caused by the bacterium Mycobacterium tuberculosis and its variants. In Brazil, the National Program for the Control and Eradication of Brucellosis and Tuberculosis in cattle, is responsible for diagnosing and the correctly allocate positive animals, but there is still a lack of definitive diagnosis of the disease. This study described the use of five diagnostic tools that can be used, preferably together, for the confirmation of suspected cases. These tools included the clinical examination comparative cervical tuberculin test, macroscopic findings during the slaughtering and histopathology of the damaged tissues followed by histochemistry. We evaluated a total of 211 dairy cattle, where 15.1% (32/211) had classic clinical signs of bovine tuberculosis, 74 (35%) showed reactivity in the comparative cervical tuberculin test. Of the total number of animals, 141 (66.8%) were referred for sanitary slaughter due to legal and control issues in the outbreaks of the disease. In the follow-up of slaughtering and inspection of viscera and carcasses, 74 (52.5%) had macroscopic lesions compatible with bovine tuberculosis, while 67 (47.5%) showed no visible changes. During the inspection, fragments of lymph nodes and liver and lung parenchyma were collected from five cattle with macroscopic lesions and five with no lesions. The histopathological analysis showed numerous areas of caseous necrosis with or without central calcification and granulomatous inflammatory infiltrate. In the special staining of Ziehl-Neelsen, numerous acid-fast bacilli were evidenced in all cases.(AU)

A tuberculose é uma antropozoonose crônica de ocorrência mundial, causada pela bactéria Mycobacterium tuberculosis e suas variantes. No Brasil existe o Programa Nacional de Controle e Erradicação da Brucelose e Tuberculose em bovinos que viabiliza o diagnóstico e a destinação correta dos animais positivos, porém ainda há carência quanto ao diagnóstico da doença. Assim, este trabalho descreve a utilização de cinco ferramentas diagnósticas para a confirmação de casos suspeitos de tuberculose. As ferramentas utilizadas compreenderam o exame clínico, teste tuberculínico cervical comparativo, os achados macroscópicos durante o abate sanitário e a histopatologia dos tecidos lesados seguido de histoquímica. O estudo avaliou um total de 211 bovinos leiteiros, dos quais 15,1% (32/211) apresentaram sinais clínicos clássicos de tuberculose bovina, 35,1% (74/211) apresentaram reatividade no teste tuberculínico cervical comparativo, e 143 animais (67,8%) foram encaminhados para abate sanitário devido a questões legais e de controle nos focos da doença. No acompanhamento do abate e inspeção sanitária de vísceras e carcaças verificou-se que 51,8% (74/143) dos bovinos abatidos apresentavam lesões macroscópicas compatíveis com tuberculose bovina, enquanto 48,2% (69/143) não apresentavam alterações visíveis. Durante a inspeção foram coletados fragmentos de linfonodos e parênquima de fígado e pulmão de cinco bovinos com lesões macroscópicas e de cinco sem lesões, que na análise histopatológica apresentaram numerosas áreas de necrose caseosa com ou sem calcificação central e infiltrado inflamatório granulomatoso. Na coloração de Ziehl-Neelsen foram evidenciados numerosos bacilos álcool-ácido resistentes em todos os casos. Assim, diante dos resultados obtidos verifica-se que as análises empregadas no presente estudo foram de extrema importância para o diagnóstico acurado de tuberculose em bovinos.(AU)

Bovine tuberculosis: diagnosis in dairy cattle through the association of analyzes / Tuberculose bovina: diagnóstico em bovinos leiteiros através da associação de análises

Tuberculosis is a chronic anthropozoonosis of worldwide occurrence, caused by the bacterium Mycobacterium tuberculosis and its variants. In Brazil, the National Program for the Control and Eradication of Brucellosis and Tuberculosis in cattle, is responsible for diagnosing and the correctly allocate positive animals, but there is still a lack of definitive diagnosis of the disease. This study described the use of five diagnostic tools that can be used, preferably together, for the confirmation of suspected cases. These tools included the clinical examination comparative cervical tuberculin test, macroscopic findings during the slaughtering and histopathology of the damaged tissues followed by histochemistry. We evaluated a total of 211 dairy cattle, where 15.1% (32/211) had classic clinical signs of bovine tuberculosis, 74 (35%) showed reactivity in the comparative cervical tuberculin test. Of the total number of animals, 141 (66.8%) were referred for sanitary slaughter due to legal and control issues in the outbreaks of the disease. In the follow-up of slaughtering and inspection of viscera and carcasses, 74 (52.5%) had macroscopic lesions compatible with bovine tuberculosis, while 67 (47.5%) showed no visible changes. During the inspection, fragments of lymph nodes and liver and lung parenchyma were collected from five cattle with macroscopic lesions and five with no lesions. The histopathological analysis showed numerous areas of caseous necrosis with or without central calcification and granulomatous inflammatory infiltrate. In the special staining of Ziehl-Neelsen, numerous acid-fast bacilli were evidenced in all cases.(AU)

A tuberculose é uma antropozoonose crônica de ocorrência mundial, causada pela bactéria Mycobacterium tuberculosis e suas variantes. No Brasil existe o Programa Nacional de Controle e Erradicação da Brucelose e Tuberculose em bovinos que viabiliza o diagnóstico e a destinação correta dos animais positivos, porém ainda há carência quanto ao diagnóstico da doença. Assim, este trabalho descreve a utilização de cinco ferramentas diagnósticas para a confirmação de casos suspeitos de tuberculose. As ferramentas utilizadas compreenderam o exame clínico, teste tuberculínico cervical comparativo, os achados macroscópicos durante o abate sanitário e a histopatologia dos tecidos lesados seguido de histoquímica. O estudo avaliou um total de 211 bovinos leiteiros, dos quais 15,1% (32/211) apresentaram sinais clínicos clássicos de tuberculose bovina, 35,1% (74/211) apresentaram reatividade no teste tuberculínico cervical comparativo, e 143 animais (67,8%) foram encaminhados para abate sanitário devido a questões legais e de controle nos focos da doença. No acompanhamento do abate e inspeção sanitária de vísceras e carcaças verificou-se que 51,8% (74/143) dos bovinos abatidos apresentavam lesões macroscópicas compatíveis com tuberculose bovina, enquanto 48,2% (69/143) não apresentavam alterações visíveis. Durante a inspeção foram coletados fragmentos de linfonodos e parênquima de fígado e pulmão de cinco bovinos com lesões macroscópicas e de cinco sem lesões, que na análise histopatológica apresentaram numerosas áreas de necrose caseosa com ou sem calcificação central e infiltrado inflamatório granulomatoso. Na coloração de Ziehl-Neelsen foram evidenciados numerosos bacilos álcool-ácido resistentes em todos os casos. Assim, diante dos resultados obtidos verifica-se que as análises empregadas no presente estudo foram de extrema importância para o diagnóstico acurado de tuberculose em bovinos.(AU)

Atypical granuloma formation in Mycobacterium bovis-infected calves.

Bovine tuberculosis is a chronic inflammatory disease that causes granuloma formation. Characterization of granulomatous lesions of Mycobacterium bovis (M. bovis) experimentally infected cattle has helped to better understand the pathogenesis of this disease. However, few studies have described granulomas found in M. bovis naturally infected cattle. The aim of this work was to examine granulomas from Holstein-Friesian cattle naturally infected with M. bovis from a dairy basin located in the central region of Mexico. Tissue samples from thirty-two cattle with lesions suggestive of tuberculosis were collected post-mortem. Fifteen of the 32 sampled animals (46.8%) were 4 months of age or younger (calves), whereas the rest (53.2%, 17/32) were over one year old (adults). Macroscopic lesions suggestive of tuberculosis were found in the mediastinal lymph node chain of all animals (32/32). From the 1,143 granulomatous lesions that were microscopically analyzed, 34.6% (396/1143) were collected from adult animals and subsequently classified according to the nomenclature suggested by Wangoo et al., 2005. Surprisingly, lesions from calf tissues showed an atypical pattern which could not be fitted into the established developmental stages of this classification. Granulomatous lesions found in calves covered most of the affected organ, histologically showed large necrotic areas with central calcification, absence of a connective tissue capsule, and few giant cells. Also, there was a higher percentage of lesions with acid-fast bacilli (AFB) when compared to studied granulomas in adults. Growth of Mycobacterium spp was detected in 11 bacteriological tissue cultures. Genotypic identification of M. bovis was performed by DNA extraction from bacterial isolates, formalin-fixed and paraffin-embedded (FFPE) tissues and samples without bacterial isolation. M. bovis was detected by PCR in 84.3% (27/32) of the studied cases; whereas other AFB were observed in tissues of the remaining sampled animals (5/32). Our results describe atypical granuloma formation in calves 4 months of age or younger, naturally infected with M. bovis. These findings contribute to better understanding the physiopathology of M. bovis infection in cattle.

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

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.(AU)

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.(AU)

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

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.(AU)

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.(AU)

Association between spoligotype-VNTR types and virulence of Mycobacterium bovis in cattle.

Mycobacterium bovis is the causative agent of bovine tuberculosis, a disease that affects approximately 5% of Argentine cattle. The aim of this research was to study if it is possible to infer the degree of virulence of different M. bovis genotypes based on scorified observations of tuberculosis lesions in cattle. In this study, we performed association analyses between several parameters with tuberculosis lesions: M. bovis genotype, degree of progression of tuberculosis, and animal age. For this purpose, the genotype was determined by spoligotyping and the degree of bovine tuberculosis gross lesion was quantified with a score based on clinical observations (number, size, and location of granulomas along with histopathologic features). This study was performed with naturally infected cattle of slaughterhouses from three provinces in Argentina. A total of 265 M. bovis isolates were obtained from 378 pathological lesion samples and 192 spoligotyping and VNTR (based on ETR sequences) typing patterns were obtained. SB0140 was the most predominant spoligotype, followed by SB0145. The spoligotype with the highest lesion score was SB0273 (median score of 27 ± 4.46), followed by SB0520 (18 ± 5.8). Furthermore, the most common spoligotype, SB0140, had a median score of 11 ± 0.74. Finally, the spoligotype with the lowest score was SB0145 (8 ± 1.0). ETR typing of SB0140, SB0145, SB0273, and SB0520 did not subdivide the lesion scores in those spoligotypes. In conclusion, SB0273 and SB0520 were the spoligotypes with the strongest association with hypervirulence and both spoligotypes were only found in Río Cuarto at the south of Córdoba province. Interestingly, there is no other report of any of these spoligotyes in Latin America.

Tuberculose cerebral bovina no oeste do Paraná: relato de caso / Tuberculosis bovine brain in the west of Parana: Case report

A Holstein heifer was attended in Palotina/PR to be in lateral recumbency and loss of appetite, excitability, nystagmus, trismus, opisthotonos, bilateral exophthalmos, opacity on right corneal, prolapse of third eyelid, congestion of scleral vessels, spasticity of the four members and increasing the volume of bilateral metacarpophalangeal and carpal joint. Euthanasia was performed and the macroscopic examination revealed injuries mainly at the base of the brain stem, small nodules in the ventricles and thickening of meninges. By microscopy was noted infl ammatory granulomatous infi ltrate with giant Langhans cells and various mineralization foci. The staining test for ZiehlNielsen resistant acid-fast bacilli was positive and confi rmed the cerebral tuberculosis, which is the fi rst report of this form of the disease in western Parana.

Uma novilha da raça holandesa foi atendida em Palotina/PR por estar em decúbito lateral e com inapetência, excitabilidade, nistagmo, trismo, opistótono, exoftalmia bilateral, opacidade da córnea direita, prolapso de terceira pálpebra, congestão dos vasos da esclera, espasticidade dos quatro membros e aumento de volume da articulação carpiana e metacarpofalangeana bilateral. Foi realizada eutanásia e o exame macroscópico revelou lesões principalmente na base do tronco encefálico, pequenos nódulos nos ventrículos e espessamento de meninges. À microscopia notou-se infiltrado inflamatório granulomatoso com células gigantes tipo Langhans e vários focos de mineralização. O exame de coloração para bacilos álcool-ácido resistentes Ziehl-Nielsen teve resultado positivo e confirmou a tuberculose cerebral, sendo este o primeiro relato desta forma da doença no oeste do Paraná.

Tuberculose cerebral bovina no oeste do Paraná: relato de caso / Tuberculosis bovine brain in the west of Parana: Case report

A Holstein heifer was attended in Palotina/PR to be in lateral recumbency and loss of appetite, excitability, nystagmus, trismus, opisthotonos, bilateral exophthalmos, opacity on right corneal, prolapse of third eyelid, congestion of scleral vessels, spasticity of the four members and increasing the volume of bilateral metacarpophalangeal and carpal joint. Euthanasia was performed and the macroscopic examination revealed injuries mainly at the base of the brain stem, small nodules in the ventricles and thickening of meninges. By microscopy was noted infl ammatory granulomatous infi ltrate with giant Langhans cells and various mineralization foci. The staining test for ZiehlNielsen resistant acid-fast bacilli was positive and confi rmed the cerebral tuberculosis, which is the fi rst report of this form of the disease in western Parana.(AU)

Uma novilha da raça holandesa foi atendida em Palotina/PR por estar em decúbito lateral e com inapetência, excitabilidade, nistagmo, trismo, opistótono, exoftalmia bilateral, opacidade da córnea direita, prolapso de terceira pálpebra, congestão dos vasos da esclera, espasticidade dos quatro membros e aumento de volume da articulação carpiana e metacarpofalangeana bilateral. Foi realizada eutanásia e o exame macroscópico revelou lesões principalmente na base do tronco encefálico, pequenos nódulos nos ventrículos e espessamento de meninges. À microscopia notou-se infiltrado inflamatório granulomatoso com células gigantes tipo Langhans e vários focos de mineralização. O exame de coloração para bacilos álcool-ácido resistentes Ziehl-Nielsen teve resultado positivo e confirmou a tuberculose cerebral, sendo este o primeiro relato desta forma da doença no oeste do Paraná.(AU)

Estudo anatomopatológico em tecidos condenados pelo Serviço De Inspeção Federal (SIF) por suspeita de tuberculose / Anatomopathological study in condemned tissues under suspicious of tuberculosis by the Brazilian food Safety and Inspection Service

A metodologia utilizada pela inspeção animal na linha de abate tem gerado controvérsias entre o que deve ser condenado e liberado ao consumo. Acredita-se que órgãos e carcaças estejam sendo condenados por tuberculose sem apresentarem lesão microscópica compatível com tal patologia. O objetivo deste trabalho foi identificar as lesões microscópicas encontradas em amostras de linfonodos condenados por tuberculose pelo SIF (Serviço de Inspeção Federal). Foram coletadas 61 amostras condenadas pelo SIF por suspeita de tuberculose. Os linfonodos foram fixados em formalina tamponada 10%, recortados, processados rotineiramente e incluídos em parafina. Na avaliação realizada pela coloração de Hematoxilina e Eosina (H&E), observaram-se 55 granulomas tuberculóides e três granulomas actinomicóides, um linfonodo apresentando cristais com leve reação inflamatória e dois hiperplasia linfóide. Concluímos, portanto, que o índice de acerto de condenação de lesões de linfonodos por tuberculose foi satisfatório, entretanto poderia ser melhorado.

The methodology used during the Animal Standard Meat Inspection (SIS) has been controversial in the decision to condemn or approve materials for human consumption. The aim of this study was to identify the microscopic lesions found in lymph node tissues condemned for tuberculosis by the Brazilian SIS. Sixty-one condemned lymph nodes were collected, fixed in buffered formalin 10% and submitted to histological processing. By evaluating the condemned tissues with H&E stain, 55 tuberculoid granuloma were found, as well as three actinomycosis granuloma, 2 presenting lymphoid hyperplasia and 1 crystal deposition. The misdiagnosis rate in the samples condemned due to tuberculosis was satisfactory, however it can be improved.

Estudo anatomopatológico em tecidos condenados pelo Serviço De Inspeção Federal (SIF) por suspeita de tuberculose / Anatomopathological study in condemned tissues under suspicious of tuberculosis by the Brazilian food Safety and Inspection Service

A metodologia utilizada pela inspeção animal na linha de abate tem gerado controvérsias entre o que deve ser condenado e liberado ao consumo. Acredita-se que órgãos e carcaças estejam sendo condenados por tuberculose sem apresentarem lesão microscópica compatível com tal patologia. O objetivo deste trabalho foi identificar as lesões microscópicas encontradas em amostras de linfonodos condenados por tuberculose pelo SIF (Serviço de Inspeção Federal). Foram coletadas 61 amostras condenadas pelo SIF por suspeita de tuberculose. Os linfonodos foram fixados em formalina tamponada 10%, recortados, processados rotineiramente e incluídos em parafina. Na avaliação realizada pela coloração de Hematoxilina e Eosina (H&E), observaram-se 55 granulomas tuberculóides e três granulomas actinomicóides, um linfonodo apresentando cristais com leve reação inflamatória e dois hiperplasia linfóide. Concluímos, portanto, que o índice de acerto de condenação de lesões de linfonodos por tuberculose foi satisfatório, entretanto poderia ser melhorado.(AU)

The methodology used during the Animal Standard Meat Inspection (SIS) has been controversial in the decision to condemn or approve materials for human consumption. The aim of this study was to identify the microscopic lesions found in lymph node tissues condemned for tuberculosis by the Brazilian SIS. Sixty-one condemned lymph nodes were collected, fixed in buffered formalin 10% and submitted to histological processing. By evaluating the condemned tissues with H&E stain, 55 tuberculoid granuloma were found, as well as three actinomycosis granuloma, 2 presenting lymphoid hyperplasia and 1 crystal deposition. The misdiagnosis rate in the samples condemned due to tuberculosis was satisfactory, however it can be improved.(AU)

Mycobacterium bovis Δmce2 double deletion mutant protects cattle against challenge with virulent M. bovis.

A Mycobacterium bovis strain deleted in mce2A and mce2B genes (M. bovis Δmce2) was tested as an experimental vaccine in cattle challenged with a virulent M. bovis strain. Three-and-a-half-month old calves (n = 5 to 6 per group) were vaccinated and challenged with a virulent strain of M. bovis by the intratracheal route 9 weeks after vaccination. A non-vaccinated group and a group vaccinated with BCG were included as controls. Blood samples were collected to measure IFN-γ by an interferon-gamma release assay (IGRA), cytometry and cytokine responses of bovine purified protein derivative (PPD) restimulated peripheral blood mononuclear cells (PBMCs). The IGRA test showed IFN-γ values similar to pre-vaccination except for the animals vaccinated with M. bovis Δmce2, where a significant increase was observed at 30 days post-vaccination. The expression of IL-2R on CD4(+) cells in response to PPD from the animals vaccinated with Δmce2 increased at 15 days post-vaccination compared to cells from non-vaccinated group. Vaccination of cattle with M. bovis Δmce2 induced the highest (P < 0.05) expression of IFN-γ and IL-17 mRNA upon PPD stimulation of PBMCs compared to vaccination with BCG or that for the non-vaccinated group. There was a weak positive correlation between the production of these proinflammatory cytokines post-vaccination and reduced pathology scores post-challenge. The animals were euthanized and necropsied 100 days after challenge. The group vaccinated with M. bovis Δmce2 displayed a significantly lower histopathological score for lesions in lungs and pulmonary lymph nodes than for the other groups (P < 0.05). A marked positive reaction to tuberculin intradermal test was observed post-vaccination in animals vaccinated with M. bovis Δmce2 compared to those vaccinated with BCG or the non-vaccinated group. In contrast, after challenge, non-vaccinated animals had greater skin test responses than the vaccinated animals. In summary, M. bovis Δmce2 is a promising vaccine candidate to control M. bovis pathogenesis in cattle.

Knockout mutation of p27-p55 operon severely reduces replication of Mycobacterium bovis in a macrophagic cell line and survival in a mouse model of infection.

Integrity of p27-p55 operon has been demonstrated to be crucial for replication of Mycobacterium tuberculosis, the main agent of human tuberculosis, in the mouse model of infection. However, the individual contribution of each gene of the operon for the virulence of pathogenic Mycobacterium spp. still remains unclear. The operon is formed by two genes, p27 and p55. p27 gene encodes a lipoprotein that binds triacylated glycolipids and modulates the host immune responses by inhibiting the MHC-II Ag processing. Besides, p55 encodes an efflux pump that, together with P27, is involved in resistance to drugs. In this study, we evaluated the individual contribution of P27 and P55 to the virulence of Mycobacterium bovis, the etiological agent for bovine tuberculosis. Knockout mutation of p27-p55 operon in M. bovis severely decreased the virulence of the bacteria when assessed in a progressive model of pulmonary tuberculosis in Balb/c mice. In addition, the mutant strain showed poor replication in a murine macrophagic cell line. Virulence and intracellular replication were only restored when the mutant strain was complemented with a copy of the whole operon. The reintroduction of p55 into the mutant strain partially restored the virulence of the bacteria while no complementation was achieved with p27 individual gene. 

Identification of proteins from tuberculin purified protein derivative (PPD) by LC-MS/MS.

The tuberculin purified protein derivative (PPD) is a widely used diagnostic antigen for tuberculosis, however it is poorly defined. Most mycobacterial proteins are extensively denatured by the procedure employed in its preparation, which explains previous difficulties in identifying constituents from PPD to characterize their behaviour in B- and T-cell reactions. We here described a proteomics-based characterization of PPD from several different sources by LC-MS/MS, which combines the solute separation power of HPLC, with the detection power of a mass spectrometer. The technique is able to identify proteins from complex mixtures of peptide fragments. A total of 171 different proteins were identified among the four PPD samples (two bovine PPD and two avium PPD) from Brazil and UK. The majority of the proteins were cytoplasmic (77.9%) and involved in intermediary metabolism and respiration (24.25%) but there was a preponderance of proteins involved in lipid metabolism. We identified a group of 21 proteins that are present in both bovine PPD but were not detected in avium PPD preparation. In addition, four proteins found in bovine PPD are absent in Mycobacterium bovis BCG vaccine strain. This study provides a better understanding of the tuberculin PPD components leading to the identification of additional antigens useful as reagents for specific diagnosis of tuberculosis.

Tuberculose bovina: saúde animal e saúde pública / Bovine tuberculosis: animal and human health / Tuberculosis bovina: salud animal y salud pública

tuberculose é uma doença infecto-contagiosa, de abrangência mundial, que afeta seres humanos e outras diversas espécies animais. A principal espécie infectante para os bovinos é o Mycobacterium bovis, o qual é endêmico no Brasil e causa prejuízos econômicos, por redução da produtividade do rebanho e perdas de carcaças no frigorífico. Por se tratar de uma zoonose, nos seres humanos pode causar uma doença clinica e patologicamente indistinguível da infecção por Mycobacterium tuberculosis. Apesar da carência de informações sobre a relevância da tuberculose bovina para os seres humanos, o aumento do número de pacientes imunossuprimidos no país, principalmente pelo vírus da imunodeficiência humana, causa preocupação em relação a doenças causadas por espécies atípicas. A espécie M. bovis é transmitida principalmente através da via aerógena para bovinos e através do consumo de leite e derivados crus para pessoas. A imunidade do hospedeiro contra os bacilos da tuberculose envolve uma variedade de eventos celulares, podendo resultar em apresentações clínicas e patologia diferenciada, variando desde infecções autolimitantes até doença sistêmica grave. O diagnóstico em bovinos é realizado principalmente por tuberculinização. Entretanto, é importante realizar a identificação laboratorial da espécie causadora da doença. Embora programas de controle e erradicação tenham sido implantados apenas na última década no país, o controle da tuberculose é fundamental para saúde pública, saúde animal e produção animal.(AU)

Tuberculosis is a worldwide infectious disease that affects human beings and several other animal species. The main specie infecting bovine is Mycobacterium bovis, which is endemic in Brazil and causes economic losses due to productivity reduction and losses at the slaughterhouse. Since tuberculosis is a zoonosis, M. bovis may infect human beings, causing a clinically or pathologically disease indistinguishable from M. tuberculosis infection. Despite the lack of information about bovine tuberculosis relevance to human beings, immunosuppressed pacients increasing in the country, mainly due to human immunodeficiency virus, causes concern about diseases caused by atypical species. Mycobacterium bovis specie is mainly transmitted by respiratory route to cattle and by raw milk and other products to human beings. The host immunity against tuberculosis bacillus includes a variety of cellular events, and may result in differentiated clinical and pathology results, ranging from self-limiting infection to severe systemic disease. Bovine diagnosis is mainly by tuberculin skin test, however, differentiate the infecting specie is important. Although control and eradication programs have been implanted only in the last decade in the country, tuberculosis control is essential for both public and animal health.(AU)

La tuberculosis es una enfermedad infecto-contagiosa, de distribución mundial, que afecta seres humanos y otras diversas especies animales. La principal especie infecciosa para los bovinos es el Mycobacterium bovis, la cual es endémica en Brasil y causa pérdidas económicas, por reducción de la productividad del rebaño y pérdidas de carcasas en el frigorífico. Además, por ser una zoonosis, en los seres humanos puede causar una enfermedad clínica y patológicamente indistinguible de la infección por M. tuberculosis. A pesar de la falta de información sobre la importancia de la tuberculosis bovina para los seres humanos, el aumento del número de pacientes inmune suprimidos en el país, principalmente por el virus de la inmunodeficiencia humana, causa preocupación en relación a enfermedades causadas por especies atípicas. La especie M. bovis es transmitida principalmente a través de la vía respiratoria para bovinos y a través del consumo de leche y derivados crudos para personas. La inmunidad del hospedador contra los bacilos de la tuberculosis incluye una variedad de eventos celulares, los cuales pueden resultar en presentaciones clínicas y patología diferenciada, variando desde infecciones autolimitantes hasta enfermedad sistémica grave. El diagnóstico en bovinos se realiza principalmente a través de prueba de la tuberculina. Sin embargo, es importante realizar la identificación en laboratorio de la especie causante de la enfermedad. Aunque programas de control y erradicación hayan sido implementados en la última década en el país, el control de tuberculosis es fundamental para la salud pública, salud animal y producción animal.(AU)

Tuberculose bovina: saúde animal e saúde pública / Bovine tuberculosis: animal and human health / Tuberculosis bovina: salud animal y salud pública

A tuberculose é uma doença infecto-contagiosa, de abrangência mundial, que afeta seres humanos e outras diversas espécies animais. A principal espécie infectante para os bovinos é o Mycobacterium bovis, o qual é endêmico no Brasil e causa prejuízos econômicos, por redução da produtividade do rebanho e perdas de carcaças no frigorífico. Por se tratar de uma zoonose, nos seres humanos pode causar uma doença clinica e patologicamente indistinguível da infecção por Mycobacterium tuberculosis. Apesar da carência de informações sobre a relevância da tuberculose bovina para os seres humanos, o aumento do número de pacientes imunossuprimidos no país, principalmente pelo vírus da imunodeficiência humana, causa preocupação em relação a doenças causadas por espécies atípicas. A espécie M. bovis é transmitida principalmente através da via aerógena para bovinos e através do consumo de leite e derivados crus para pessoas. A imunidade do hospedeiro contra os bacilos da tuberculose envolve uma variedade de eventos celulares, podendo resultar em apresentações clínicas e patologia diferenciada, variando desde infecções autolimitantes até doença sistêmica grave. O diagnóstico em bovinos é realizado principalmente por tuberculinização. Entretanto, é importante realizar a identificação laboratorial da espécie causadora da doença. Embora programas de controle e erradicação tenham sido implantados apenas na última década no país, o controle da tuberculose é fundamental para saúde pública, saúde animal e produção animal.

Tuberculosis is a worldwide infectious disease that affects human beings and several other animal species. The main specie infecting bovine is Mycobacterium bovis, which is endemic in Brazil and causes economic losses due to productivity reduction and losses at the slaughterhouse. Since tuberculosis is a zoonosis, M. bovis may infect human beings, causing a clinically or pathologically disease indistinguishable from M. tuberculosis infection. Despite the lack of information about bovine tuberculosis relevance to human beings, immunosuppressed pacients increasing in the country, mainly due to human immunodeficiency virus, causes concern about diseases caused by atypical species. Mycobacterium bovis specie is mainly transmitted by respiratory route to cattle and by raw milk and other products to human beings. The host immunity against tuberculosis bacillus includes a variety of cellular events, and may result in differentiated clinical and pathology results, ranging from self-limiting infection to severe systemic disease. Bovine diagnosis is mainly by tuberculin skin test, however, differentiate the infecting specie is important. Although control and eradication programs have been implanted only in the last decade in the country, tuberculosis control is essential for both public and animal health.

La tuberculosis es una enfermedad infecto-contagiosa, de distribución mundial, que afecta seres humanos y otras diversas especies animales. La principal especie infecciosa para los bovinos es el Mycobacterium bovis, la cual es endémica en Brasil y causa pérdidas económicas, por reducción de la productividad del rebaño y pérdidas de carcasas en el frigorífico. Además, por ser una zoonosis, en los seres humanos puede causar una enfermedad clínica y patológicamente indistinguible de la infección por M. tuberculosis. A pesar de la falta de información sobre la importancia de la tuberculosis bovina para los seres humanos, el aumento del número de pacientes inmune suprimidos en el país, principalmente por el virus de la inmunodeficiencia humana, causa preocupación en relación a enfermedades causadas por especies atípicas. La especie M. bovis es transmitida principalmente a través de la vía respiratoria para bovinos y a través del consumo de leche y derivados crudos para personas. La inmunidad del hospedador contra los bacilos de la tuberculosis incluye una variedad de eventos celulares, los cuales pueden resultar en presentaciones clínicas y patología diferenciada, variando desde infecciones autolimitantes hasta enfermedad sistémica grave. El diagnóstico en bovinos se realiza principalmente a través de prueba de la tuberculina. Sin embargo, es importante realizar la identificación en laboratorio de la especie causante de la enfermedad. Aunque programas de control y erradicación hayan sido implementados en la última década en el país, el control de tuberculosis es fundamental para la salud pública, salud animal y producción animal.

High-level expression of NRAMP1 in peripheral blood cells and tuberculous granulomas from Mycobacterium bovis-infected bovines.

By Western blotting, we demonstrate high-level expression of NRAMP1 proteins in peripheral blood cells and granulomas of Mycobacterium bovis-infected bovines. Immunohistochemistry of granulomatous lesions showed heavily labeled epithelioid macrophages and Langhans cells. These data suggest that M. bovis infection enhances NRAMP1 expression and that active tuberculosis can occur despite this response.