O gênero Mycobacterium compreende inúmeras espécies estudadas por sua capacidade de causar doenças. Surtos de tuberculose e micobacteriose em zoológicos, aquários, centros de pesquisa e criadouros evidenciaram a importância desses organismos em relação à saúde de animais silvestres mantidos em cativeiro. Duzentas e onze amostras, de 101 animais entre mamíferos, aves, répteis, anfíbios e peixes, foram submetidas à cultura. Foram isolados M. tuberculosis, M. bovis e 13 espécies de MNT associadas à doença. A natureza crônica dessas doenças, seu diagnóstico ante mortem limitado, juntamente com as características de interação social de cada espécie e o confinamento, favorecem a transmissão das micobactérias pelo contato com humanos ou outros animais infectados. Apesar de a presença de M. tuberculosis e M. bovis em instituições brasileiras já ter sido constatada, o presente estudo descreveu infecções em espécies animais utilizadas para consumo humano, e em espécies conhecidas pela suscetibilidade, mas que ainda não foram documentadas no Brasil. Doença causada por MNT foi relatada, mas sua importância deve ser considerada de acordo com a espécie animal acometida / Mycobacterium genus comprises several species, which have been studied for their capacity to cause diseases. Outbreaks of tuberculosis and mycobacteriosis in zoos, aquariums, research and breeding facilities have shown the importance of these organisms to health of wildlife animal maintained in captivity. Two hundred and eleven samples, from 101 animals among mammal, birds, reptiles, amphibians and fishes, were cultured. Mycobacterium tuberculosis, M. bovis and 13 species of non-tuberculous mycobacteria associated to diseases were isolated. The chronical nature of tuberculosis and mycobacteriosis, their limited ante mortem diagnosis, along with social behavior of each species and confinement, favor mycobacterial transmission through contact with infected humans and other animals. Despite the presence of M. tuberculosis and M. bovis has already been observed in Brazilian institutions, the present study described infections in animal species destined to human consumption, and species known for their susceptiblility, yet not documented in Brazil. Disease caused by non-tuberculous mycobacteria was reported, though its importance must be considered according to the animal species affected

O gênero Mycobacterium compreende inúmeras espécies estudadas por sua capacidade de causar doenças. Surtos de tuberculose e micobacteriose em zoológicos, aquários, centros de pesquisa e criadouros evidenciaram a importância desses organismos em relação à saúde de animais silvestres mantidos em cativeiro. Duzentas e onze amostras, de 101 animais entre mamíferos, aves, répteis, anfíbios e peixes, foram submetidas à cultura. Foram isolados M. tuberculosis, M. bovis e 13 espécies de MNT associadas à doença. A natureza crônica dessas doenças, seu diagnóstico ante mortem limitado, juntamente com as características de interação social de cada espécie e o confinamento, favorecem a transmissão das micobactérias pelo contato com humanos ou outros animais infectados. Apesar de a presença de M. tuberculosis e M. bovis em instituições brasileiras já ter sido constatada, o presente estudo descreveu infecções em espécies animais utilizadas para consumo humano, e em espécies conhecidas pela suscetibilidade, mas que ainda não foram documentadas no Brasil. Doença causada por MNT foi relatada, mas sua importância deve ser considerada de acordo com a espécie animal acometida / Mycobacterium genus comprises several species, which have been studied for their capacity to cause diseases. Outbreaks of tuberculosis and mycobacteriosis in zoos, aquariums, research and breeding facilities have shown the importance of these organisms to health of wildlife animal maintained in captivity. Two hundred and eleven samples, from 101 animals among mammal, birds, reptiles, amphibians and fishes, were cultured. Mycobacterium tuberculosis, M. bovis and 13 species of non-tuberculous mycobacteria associated to diseases were isolated. The chronical nature of tuberculosis and mycobacteriosis, their limited ante mortem diagnosis, along with social behavior of each species and confinement, favor mycobacterial transmission through contact with infected humans and other animals. Despite the presence of M. tuberculosis and M. bovis has already been observed in Brazilian institutions, the present study described infections in animal species destined to human consumption, and species known for their susceptiblility, yet not documented in Brazil. Disease caused by non-tuberculous mycobacteria was reported, though its importance must be considered according to the animal species affected

Submitted by Cássio da Cruz Nogueira (cassionogueirakk@gmail.com) on 2017-01-30T13:48:19Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_AvaliacaoAtividadeExtrato.pdf: 2549621 bytes, checksum: 323b92687db50fd9c3216ceeb94a8fec (MD5) / Approved for entry into archive by Edisangela Bastos (edisangela@ufpa.br) on 2017-02-01T11:55:12Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_AvaliacaoAtividadeExtrato.pdf: 2549621 bytes, checksum: 323b92687db50fd9c3216ceeb94a8fec (MD5) / Made available in DSpace on 2017-02-01T11:55:12Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_AvaliacaoAtividadeExtrato.pdf: 2549621 bytes, checksum: 323b92687db50fd9c3216ceeb94a8fec (MD5) Previous issue date: 2014-10 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico / Chenopodium ambrosioides e Eucalyptus alba são espéciesutilizadas na medicina popular para o tratamento da tuberculose e expectoração. O presente trabalho teve o objetivo de determinar os parâmetros físico-químicos e microbiológicos, definir o perfil fitoquímico e avaliar as atividades antimicobacteriana, citotóxica, imunomodulatória e toxicológica in vivo dos extratos e frações. Na avaliação físico-química e microbiológica dos extratos de C. ambrosioides e de E. alba, foram encontrados parâmetros em conformidade com a literatura específica para plantas medicinais. A prospecção fitoquímica dos extratos revelou a presença de saponinas, esteroides, triterpenoides, fenóis e taninos, o extrato de C. ambrosioides apresentou também proteínas e aminoácidos, enquanto o extrato de E. alba foi positivo para ácidos orgânicos e lactonas. Os resultados do método da microdiluição em caldo e da microdiluição utilizando Alamar Blue evidenciaram moderadas atividades frente à Mycobacterium fortuitum do extrato de E. alba e da fração acetato de etila (FAcE) de E. alba; a fração clorofórmica (FCl) de C. ambrosioides e a FAcE de E. alba foram moderadamente ativas frente à Mycobacterium tuberculosis. A atividade citotóxica, avaliada pelo método do MTT, demonstrou que os extratos não reduziram a viabilidade celular nas concentrações testadas. No ensaio de imunomodulação, o extrato de E.alba apresentou potencial efeito anti-inflamatório, pelos métodos do inibição da produção de NO e TNFα. Não foram detectados sinais de toxicidade oral aguda dos extratos na dose de 2500 mg/kg em camundongos. Estes resultados sugerem o potencial antimicobacteriano da FAcE de E. alba e imunomodulador do extrato de E. alba, e podem servir como subsídio para estudos futuros com vista no isolamentode compostos ativos e elucidação de seus mecanismos. / Chenopodium ambrosioides and Eucalyptus alba are species used in folk medicine for the treatment of tuberculosis and sputum. This study aimed to determine the physical-chemical, microbiological parameters, define phytochemical screening and evaluate antimycobacterial, cytotoxicity, immunomodulation, and toxicity in vivo activities of extracts and fractions. In physical-chemical and microbiological evaluation of C.ambrosioides and E. alba extracts, were found parameters in accordance with the specific literature for medicinal plants. The phytochemical screening of the extracts revealed the presence of saponins, steroids, triterpenoids, phenols and tannins, C. ambrosioides extract also showed proteins and amino acids, while the E. alba extract was positive for organic acids and lactones. The results of broth microdilution assay and microplate alamar blue assay showed moderates activities against the Mycobacterium fortuitum of E. alba extract and ethyl acetate fraction (FAcE) of E. alba; the chloroform fraction (FCl) of C.ambrosioides and the ethyl acetate fraction (FacEA) of E. alba were moderately active against the Mycobacterium tuberculosis. The cytotoxic activity, evaluated by MTT method, showed that the extracts did not reduce cell viability in the concentrations tested. In the immunomodulation assay, E. alba extract presented potential anti-inflammatory effect, by the methods of inhibition the production of NO and TNFα. No signs of acute oral toxicity of the extracts at a dose of 2500 mg/kg in mice were detected. These results suggest the potential antimycobacterial the FAcE of E. alba and immunomodulatory of E. alba extract and can serve as a resource for future studies, aimed at isolation of active compound and elucidation of their mechanisms of action.

Polycyclic aromatic hydrocarbons, produced by anthropological and natural activities, are hazardous through formation of oxidative radicals and DNA adducts. Growth of Mycobacterium sp. strain KMS, isolated from a contaminated soil, on the model hydrocarbon pyrene induced specific proteins. My work extends the study of isolate KMS to the gene level to understand the pathways and regulation of pyrene utilization. Genes encoding pyrene-induced proteins were clustered on a 72 kb section on the KMS chromosome but some also were duplicated on plasmids. Skewed GC content and presence of integrase and transposase genes suggested horizontal transfer of pyrene-degrading gene islands that also were found with high conservation in five other pyrene-degrading Mycobacterium isolates. Transcript analysis found both plasmid and chromosomal genes were induced by pyrene. These processes may enhance the survival of KMS in hydrocarbon-contaminated soils when other carbon sources are limited. KMS also grew on benzoate, confirming the functionality of an operon containing genes distinct from those in other benzoate-degrading bacteria. Growth on benzoate but not on pyrene induced a gene, benA, encoding a benzoate dioxygenase α-subunit, but not the pyrene-induced nidA encoding a pyrene dioxygenase α-subunit; the differential induction correlated with differences in promoter sequences. Diauxic growth occurred when pyrene cultures were amended with benzoate or acetate, succinate, or fructose, and paralleled delayed expression of nidA. Single phase growth and normal expression of benA was observed for benzoate single and mixed cultures. The nidA promoters had potential cAMP-CRP binding sites, suggesting that cAMP could be involved in carbon repression of pyrene metabolism. Growth on benzoate and pyrene requires gluconeogenesis. Intermediary metabolism in isolate KMS involves expression from genes encoding a novel malate:quinone oxidoreductase and glyoxylate shunt enzymes. Generation of C3 structures involves transcription of genes encoding malic enzyme, phosphoenolpyruvate carboxykinase, and phosphoenolpyruvate synthase. Carbon source modified the transcription patterns for these genes. My findings are the first to show duplication of pyrene-degrading genes on the chromosome and plasmids in Mycobacterium isolates and expression from a unique benzoate-degrading operon. I clarified the routes for intermediary metabolism leading to gluconeogenesis and established a potential role for cAMP-mediated catabolite repression of pyrene utilization.

Submitted by (edna.saturno@ufrpe.br) on 2016-10-07T14:44:50Z No. of bitstreams: 1 Flavia Figueiraujo Jabour.pdf: 2776762 bytes, checksum: c6f5ab38588135603c3ae6a6a04e7a81 (MD5) / Made available in DSpace on 2016-10-07T14:44:50Z (GMT). No. of bitstreams: 1 Flavia Figueiraujo Jabour.pdf: 2776762 bytes, checksum: c6f5ab38588135603c3ae6a6a04e7a81 (MD5) Previous issue date: 2013-02-28 / Tuberculosis, paratuberculosis and caseous lymphadenitis are important granulomatous diseases that affect various animal species, including sheep and goats and determine major economic losses. These diseases have similar clinical and pathological characteristics and deserve further investigation and differentiated to facilitate its control. The objective was to study to diagnose the occurrence of granulomatous diseases such as tuberculosis, paratuberculosis and caseous lymphadenitis in sheep and goats slaughtered in a municipal slaughterhouse, State of Alagoas, Brazil. Two hundred animals had been slaughtered during four visits made to the slaughterhouse from 2009 to 2011. Among these two hundred animals, 39 sheep and 3 goats presented macroscopic lesions in several body tissues. Histopathological, immunohistochemistry and bacteriological exams were executed for diagnostic. Lesions in the gut compatible with alterations caused by Oesophagostomum sp. were found in the sheep and goats. All samples were negative in immunohistochemistry for Mycobacterium spp. In the bacteriological test, Corybacterium pseudotuberculosis was isolated from lung, lymph nodes, small and large gut of 18 sheep and from the large gut of one goat. Alterations of tuberculosis and paratuberculosis were not observed, however lesions of other economically important diseases were found, such as caseous lymphadenitis associated with alterations of oesophagostomiasis e fascioliasis. In the second studythe objective was to perform the tuberculin skin test in 275 sheep and 45 goats coming from four herds in three cities of Eastern Mesoregion of Alagoas state. We also applied a questionnaire to analyze the epidemiological profile of the properties studied. All goats were negative to the tuberculin skin test and two sheep (0.72%) were positive for two different properties (50%). Join the occurrence of tuberculosis in sheep in the state of Alagoas. Despite the lower frequency of cases there were a high number of outbreaks. This result indicates that one should intensify their efforts to control tuberculosis in these regions studied to prevent the spread of mycobacteria to other flocks of sheep and goats and similar epidemiological studies should be conducted in other regions of the state of Alagoas to investigate the occurrence of disease and your impact in sheep and goat herds. / Tuberculose, paratuberculose e linfadenite caseosa são doenças granulomatosas importantes que acometem várias espécies animais, incluindo ovinos e caprinos e determinam grandes prejuízos econômicos. Estas doenças possuem características clínico-patológicas semelhantes e merecem ser melhor investigadas e diferenciadas para facilitar seu controle. Objetivou-se estudar a ocorrência de doenças granulomatosas em ovinos e caprinos abatidos em um matadouro no Estado de Alagoas, além de determinar a ocorrência de tuberculose em ovinos e caprinos em propriedades rurais na região da Zona da Mata Alagoana. No primeiro estudo Foram estudados 200 animais abatidos nos anos de 2009 a 2011. Destes, 39 ovinos e três caprinos apresentaram lesões macroscópicas em vários tecidos. Para o diagnóstico foram realizados exames histopatológico e bacteriológico, e imunohistoquímica. Observou-se, ao exame histopatológico, lesões intestinais compatíveis com Oesophagostomum sp. em ovinos e caprinos e com fasciolose no fígado de ovinos. Na imunohistoquímica todas as amostras foram negativas para Mycobacterium spp. Ao exame bacteriológico isolou-se Corynebacterium pseudotuberculosis nos tecidos de 18 ovinos (pulmão, fígado, linfonodos, intestino delgado e intestino grosso) e de um caprino (intestino grosso). Não foi diagnosticada tuberculose e paratuberculose nos animais estudados, no entanto observou-se a ocorrência de outras doenças economicamente importantes como linfadenite caseosa associada a lesões de oesofagostomose e fasciolose. No segundo estudo objetivou-se realizar o diagnóstico da tuberculose em caprinos e ovinos em três municípios da Mesorregião do Leste Alagoano. Foram tuberculinizados 275 ovinos e 45 caprinos procedentes de quatro rebanhos. Também foram aplicados questionários para estudar o perfil epidemiológico das propriedades estudadas. Todos os caprinos foram negativos ao teste tuberculínico e dois ovinos (0,72%) foram positivos, registrando-se 50% de focos da doença. Registra-se a ocorrência da tuberculose em ovinos no estado de Alagoas. Apesar da baixa frequência de casos positivos observou-se um elevado número de focos da doença. Este resultado indica que medidas de controle devem ser intensificadas para evitar a disseminação de micobactérias a outros rebanhos de ovinos e caprinos e estudos epidemiológicos semelhantes devem ser realizados em outras regiões do estado de Alagoas para investigar a ocorrência da doença e seu impacto na criação de caprinos e ovinos. Apesar de não ter sido diagnosticada a tuberculose clínica e a paratuberculose nos animais estudados, observou-se a ocorrência de outras doenças economicamente importantes como linfadenite caseosa associada a lesões de oesofagostomose e fasciolose nos animais abatidos.

Polycyclic aromatic hydrocarbons, produced by anthropological and natural activities, are hazardous through formation of oxidative radicals and DNA adducts. Growth of Mycobacterium sp. strain KMS, isolated from a contaminated soil, on the model hydrocarbon pyrene induced specific proteins. My work extends the study of isolate KMS to the gene level to understand the pathways and regulation of pyrene utilization. Genes encoding pyrene-induced proteins were clustered on a 72 kb section on the KMS chromosome but some also were duplicated on plasmids. Skewed GC content and presence of integrase and transposase genes suggested horizontal transfer of pyrene-degrading gene islands that also were found with high conservation in five other pyrene-degrading Mycobacterium isolates. Transcript analysis found both plasmid and chromosomal genes were induced by pyrene. These processes may enhance the survival of KMS in hydrocarbon-contaminated soils when other carbon sources are limited. KMS also grew on benzoate, confirming the functionality of an operon containing genes distinct from those in other benzoate-degrading bacteria. Growth on benzoate but not on pyrene induced a gene, benA, encoding a benzoate dioxygenase α-subunit, but not the pyrene-induced nidA encoding a pyrene dioxygenase α-subunit; the differential induction correlated with differences in promoter sequences. Diauxic growth occurred when pyrene cultures were amended with benzoate or acetate, succinate, or fructose, and paralleled delayed expression of nidA. Single phase growth and normal expression of benA was observed for benzoate single and mixed cultures. The nidA promoters had potential cAMP-CRP binding sites, suggesting that cAMP could be involved in carbon repression of pyrene metabolism. Growth on benzoate and pyrene requires gluconeogenesis. Intermediary metabolism in isolate KMS involves expression from genes encoding a novel malate:quinone oxidoreductase and glyoxylate shunt enzymes. Generation of C3 structures involves transcription of genes encoding malic enzyme, phosphoenolpyruvate carboxykinase, and phosphoenolpyruvate synthase. Carbon source modified the transcription patterns for these genes. My findings are the first to show duplication of pyrene-degrading genes on the chromosome and plasmids in Mycobacterium isolates and expression from a unique benzoate-degrading operon. I clarified the routes for intermediary metabolism leading to gluconeogenesis and established a potential role for cAMP-mediated catabolite repression of pyrene utilization.