Submitted by (lucia.rodrigues@ufrpe.br) on 2017-02-09T15:10:13Z No. of bitstreams: 1 Luenda de Menezes e Sa.pdf: 4392852 bytes, checksum: 551c75da448596a356470434e8984578 (MD5) / Made available in DSpace on 2017-02-09T15:10:13Z (GMT). No. of bitstreams: 1 Luenda de Menezes e Sa.pdf: 4392852 bytes, checksum: 551c75da448596a356470434e8984578 (MD5) Previous issue date: 2012-02-09 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Paratuberculosis is a disease that can cause a negative impact on national livestock. In cattle cause chronic intractable diarrhea and granulomatous enteritis, these animals become listless and lose weight gradually, decreasing or ceasing milk production. It is caused by Mycobacterium avium subspecies paratuberculosis (MAP), an alcohol-acid resistant bacilli, resistant to many environments. The objective of this study makes an epidemiological investigation of MAP infections in dairy cattle the microregion of Garanhuns, Pernambuco, Brazil. For this study blood samples were collected from 408 animals from 19 herds in 15 counties of the microregion. Serologic testing was performed by indirect enzyme immunoassay (ELISA) for antibodies against the MAP. A questionnaire was applied to all properties for analysis of risk factors, and collected geographical coordinates through a GPS (Global Position System) device to perform the spatial distribution. The prevalence of MAP infection was 2.7% (11/408, I.C. 1.4 – 4.9). The number of foci was 47.4% (9/19). In logistic regression analysis was identified as a risk factor the annual rate of births over 51 calves / year (O.R. 3.8; I.C. 1.1 – 13.1). In conclusion an MAP infection is spread among the cattle’s herds of microregion Garanhuns, Pernambuco, Brazil, and control measures based or risk factors should be implemented to reduce the number of outbreaks and consequently increase the productivity of herds. / A paratuberculose é uma enfermidade que pode causar um impacto negativo para a pecuária nacional. Nos bovinos apresenta-se sob a forma de diarreia crônica intratável que causa uma enterite granulomatosa, onde os animais tornam-se apáticos e emagrecem progressivamente, diminuindo ou até mesmo cessando a produção leiteira. É causada pelo Mycobacterium avium subespécie paratuberculosis (MAP), um pequeno bacilo álcool-ácido-resistente, considerado como um dos menores do gênero, extremamente resistente a diversos ambientes. Objetivou-se com esse trabalho realizar um inquérito epidemiológico da infecção pelo MAP em bovinos leiteiros da microrregião de Garanhuns, Pernambuco, Brasil. Para esse estudo foram coletadas amostras sanguíneas de 408 animais, provenientes de 19 rebanhos localizados em 15 municípios da microrregião. O exame sorológico foi realizado por Ensaio Imunoenzimático (ELISA) indireto para detecção de anticorpos frente ao Mycobacterium avium subsp. paratuberculosis. Em todas as propriedades, um questionário investigativo foi aplicado para análise dos fatores de risco, e as coordenadas geográficas coletadas através de um aparelho de GPS (Global Position System) para realização da distribuição espacial. A prevalência da infecção pelo MAP foi de 2,7% (11/408; I.C. 1,4 – 4,9). O número de focos foi 47,4% (9/19). Na análise de regressão logística foi identificado como fator de risco a taxa anual de nascimentos superior a 51 bezerros/ano (OR 3,8; I.C. 1,1 – 13,1). Concluiu-se que a infecção pelo MAP encontra-se distribuída entre os rebanhos bovinos leiteiros da microrregião estudada e que medidas de controle baseadas nos fatores de risco devem ser implementadas com o objetivo de reduzir o número de focos e consequentemente aumentar a produtividade dos rebanhos, minimizando assim os prejuízos causados pela enfermidade aos produtores.

La paratuberculose bovine, ou maladie de Johne, est une maladie inflammatoire intestinale chronique provoquant d’importantes pertes économiques chez les producteurs de ruminants du monde entier. Que ce soit chez la vache laitière ou de boucherie, ces pertes sont causées majoritairement par une diminution de la capacité de reproduction, la baisse de production laitière et l’amaigrissement des vaches qui perdent ainsi beaucoup de valeur à l’abattage, en plus d’être sujettes à une réforme précoce. Outre les pertes économiques, le potentiel de transmission à l’humain est un facteur non négligeable en plus d’un risque de contamination de la chaîne alimentaire. Cette maladie est causée par une bactérie intracellulaire obligatoire nommée Mycobacterium avium subspecies paratuberculosis (MAP). II n’existe actuellement aucune stratégie efficace pour combattre l’infection chez les animaux atteints. L’évolution lente de la maladie fait en sorte que les signes cliniques apparaissent tardivement, soit plusieurs années (4 à 7 ans) après l’infection initiale. Au cours de cette progression, les animaux infectés commencent à excréter le pathogène dans leur environnement. Les animaux atteints deviennent infectieux et peuvent contaminer d’autres congénères, ainsi que leur propre veau. Afin de permettre aux producteurs d’éliminer les vaches atteintes avant qu’elles n’atteignent ce stade, il s’avère important d’établir un diagnostic précoce. Actuellement, ce n’est qu’en phase sous-clinique avancée que les tests diagnostiques sont plus sensibles, soit 2 à 3 ans après le début des excrétions fécales chez les animaux infectés. L'incompréhension du manque de sensibilité des tests de dépistage et de l'évolution de cette maladie justifient les efforts de recherche dans ce domaine en vue de mieux comprendre les réponses immunitaires impliquées dans cette maladie. En effet, une meilleure connaissance des processus d’inflammation chronique pourrait aider à développer des outils diagnostiques complémentaires. Nos résultats suggèrent une dérégulation de la réponse immunitaire. Ainsi, en étudiant les composantes et caractéristiques du sang provenant de vaches infectées, il nous a été possible d’observer que les niveaux de cytokines plasmatiques telles l’interleukine 17 et l'ostéopontine se trouvent sécrétées à différents niveaux chez les vaches atteintes de paratuberculose bovine. De plus, l'analyse de la capacité de leur sérum à soutenir efficacement la prolifération des cellules mononucléées du sang périphérique révèle que le sérum de vaches infectées interfère pour atténuer la prolifération cellulaire. II semble qu’un constituant du sérum provoque une diminution de la réponse immunitaire chez les vaches malades. Les résultats offrnt une appréciation des dérèglements immunitaires provoqués par la paratuberculose bovine.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2017-05-03T21:15:13Z No. of bitstreams: 1 2016 - Daniel Guimar?es Ubiali.pdf: 7253233 bytes, checksum: ce10a8e3d4ef67ad93a0034844382ed3 (MD5) / Made available in DSpace on 2017-05-03T21:15:13Z (GMT). No. of bitstreams: 1 2016 - Daniel Guimar?es Ubiali.pdf: 7253233 bytes, checksum: ce10a8e3d4ef67ad93a0034844382ed3 (MD5) Previous issue date: 2016-10-26 / CAPES / For investigation of Mycobacterium avium subsp. paratuberculosis infection 34 buffaloes properties or ranches in Northeastern Brazil were visited for paratuberculosis diagnosis. Investigations included herd evaluations, inspection of facilities and pastures, obtainment of flock history, clinical examination of suspicious animals and collecting samples for diagnosis. Samples were obtained from 26 farms or ranches, including two slaughterhouses and a quarantine area in six states. Approximately 15,600 buffalos, including males and females of the Murrah, Mediterranean and Jafarabadi breeds as well as their crossbreeds, were evaluated for meat, dairy and mixed properties with semi-intensive or extensive regimes. For diagnostic purposes, necropsies, histopathological and immunohistochemical exams and Ziehl-Neelsen tests of fecal smears and scraped intestinal mucosa were performed. Polymerase chain reaction (PCR) methods were applied to samples of feces, milk, mesenteric nodes and intestines. This exams allowed us to identify eigth new Johne?s disease outbreaks, which, together with those previously identified by our staff, allow us to infer that the disease is being dispersed in the Brazilian Northeast, similar to what is occurring with bovine herds in other areas of the country. The increase in the number of positive farms is a consequence of the ignorance of farmers, inadequate health management, free trade of ruminants and lack of a official control program in the country. This study alerts for risk of commercialization of dairy products for human consumption, reinforces the importance of research on paratuberculosis in Brazil and contributes to the understanding of the factors that work together to increase the number of paratuberculosis cases in the Brazilian Northeast. / Com o objetivo de identificar focos e estudar a epidemiologia e o diagn?stico da paratuberculose em b?falos na regi?o Nordeste do Brasil foram realizadas visitas e ou examinados material provenientes de 34 propriedades com suspeita cl?nica da doen?a. Obtivemos material biol?gico de sete estados da regi?o Nordeste do Brasil (Maranh?o, Cear?, Para?ba, Pernambuco, Alagoas e Bahia). Dos rebanhos foi obtido o hist?rico, realizou-se avalia??o cl?nica e inspe??o das instala??es e pastagens. Para a confirma??o do diagn?stico foi coletado material para exames laboratoriais em 26 propriedades ou cria??es de b?falos, entre estes, dois matadouros e um quarenten?rio. Foram rebanhos cujo somat?rios de b?falos era de aproximadamente 15.600 b?falos, das ra?as Murrah, Mediterr?neo, Jafarabadi e seus mesti?os, com aptid?o para corte, leite ou mista, em propriedades com regimes semi-intensivo, extensivo ou extrativista. Foram realizadas 22 necropsias e coleta de material para exames histopatol?gicos e imuno-histoqu?micos, al?m de colora??o de Ziehl-Neelsen em esfrega?os de fezes, raspados de mucosa intestinal e fragmentos de linfonodo mesent?rico e de intestino que apresentavam les?es sugestivas da doen?a. Para a realiza??o da rea??o em cadeia da polimerase (PCR) foram utilizadas amostras de fezes, leite, linfonodos mesent?ricos e intestinos. Estes exames permitiram identificar oito focos de paratuberculose, o que nos permite inferir que a doen?a est? se dispersando na regi?o Nordeste do Brasil a exemplo do que est? acontecendo em outras regi?es do pa?s com o rebanho bovino. O desconhecimento da doen?a, o manejo inadequado, o com?rcio n?o regulamentado de b?falos e a falta de um programa de controle voltado para a realidade da regi?o facilitam a dispers?o do agente e s?o fatores que contribuem para o aumento do n?mero de focos no pa?s. Os resultados desta pesquisa contribuem com a epidemiologia e o diagn?stico da doen?a em b?falos e auxilia na compreens?o dos fatores que colaboram para a ocorr?ncia crescente do n?mero de casos desta doen?a.

Mycobacterium avium subsp. paratuberculosis (MAP) cause la maladie de Johne, une maladie chronique et incurable affectant les ruminants partout dans le monde. Plusieurs pays ont mis en place des programmes de contrôle afin de prévenir la transmission entre et au sein des troupeaux. Afin d’arriver à prévenir et contrôler cette maladie, une bonne compréhension des facteurs de risque impliqués dans la transmission est essentielle. Des tests diagnostiques performants et à coût abordable sont aussi nécessaires afin de détecter la présence du MAP et/ou les animaux infectés. L’objectif de la première étude était de réviser systématiquement la littérature scientifique concernant les facteurs de risque associés à la transmission du MAP aux génisses laitières. La présence d’une association significative entre les facteurs de risque concernant l’environnement néonatal, le colostrum, le lait, le logement des veaux et le contact des veaux avec le fumier de vaches adultes et la transmission du MAP a été compilée de 23 articles. Le contact des veaux avec le fumier de vaches adultes est le facteur de risque le plus important dans la transmission du MAP. L’objectif de la seconde étude était d’évaluer la relation entre le nombre d’échantillons de l’environnement positifs pour le MAP et la prévalence individuelle d’excrétion fécale dans les troupeaux laitiers entravés du Québec. Le nombre de cultures positives d’échantillons de l’environnement s’est avéré associé à la prévalence individuelle d’excrétion fécale du MAP. Une association significative a été trouvée entre la présence d’une forte charge bactérienne dans un échantillon de fumier individuel et la détection du MAP dans l’environnement. / Mycobacterium avium subsp. paratuberculosis (MAP) causes Johne’s disease, a chronic and incurable enteric disease of ruminants that has a worldwide distribution. Many countries have implemented control programs to prevent transmission between and within herds. For these to be efficient, knowledge of the risk factors involved in transmission is essential. Efficient and cost effective diagnostic tests are also necessary to detect presence of MAP. The objective of the first study was to systematically review the scientific literature concerning risk factors associated with MAP transmission to dairy calves. Presence of a significant association between risk factors concerning neonatal environment, colostrum, milk, housing of calves and contact of calves with adult cow feces and MAP transmission were recorded from 23 articles. The contact of calves with adult cow feces appeared to be the most important risk factor in MAP transmission. The 5 categories of risk factors are linked one to each other. The objective of the second study was to evaluate the relation between the number of positive samples for MAP in the environment and the prevalence of individual fecal shedding in Québec tie-stall dairy herds. The number of positive cultures of environmental samples was associated with the prevalence of individual fecal shedding of MAP. A significant association was found between presence of a heavy bacterial load in an individual fecal sample and environmental detection of MAP.

La malattia di Johne o paratubercolosi è un’enterite cronica granulomatosa provocata dal Mycobacterium avium subsp paratubercolosis (MAP), che colpisce i ruminanti ed in particolare i bovini da latte ed ha un grande impatto economico a livello mondiale. Il MAP sembra anche avere un ruolo nella malattia umana di Crohn. Tale patogeno è capace di sopravvivere molto bene all’interno dei macrofagi dell’ospite dove previene la loro attivazione, blocca l’acidificazione e la maturazione del fagosoma, e interferisce con la presentazione degli antigeni al sistema immunitario. Al fine di analizzare la complessa interazione tra l’ospite e il patogeno, è stata valutata la risposta dopo 2h, 6h, e 24h di macrofagi derivati da monociti bovini (MDM), coltivati in vitro e infettati con il ceppo L1 di MAP utilizzando un approccio di RNA-Seq. L’analisi statistica dei dati di sequenza ha mostrato un aumento del numero di geni differenzialmente espressi durante l’esperimento in risposta all’infezione. Inoltre i geni sottoespressi negli MDM infettati sono stati individuati solo a 24h post-infezione. L’analisi dei pathway ha evidenziato tre network che sono associati alla risposta immunitaria e al processo infiammatorio. Inoltre lo studio dei geni sottoespressi a 24h ha mostrato il ruolo centrale del complemento e del complesso maggiore di istocompatibilità nella patogenesi della malattia. / Johne’s disease (paratuberculosis) is a chronic granulomatous enteritis caused by Mycobacterium avium subsp paratubercolosis (MAP), affecting ruminants worldwide with a significant economic impact. MAP has also been speculated as a cause of human Crohn’s disease. MAP is a pathogen highly adapted for survival within host macrophages due to the organism's capacity to prevent macrophage activation, block phagosome acidification and maturation, and attenuate presentation of antigens to the immune system. The consequence is a very long silent infection and subclinical phases. To decipher the complex interaction between host and MAP, the response of in vitro bovine monocyte-derived macrophages (MDM) after 2h, 6h and 24h of infection with L1 strain of MAP was explored using RNA-Seq approach. Statistical analysis of sequence data revealed an increasing number of differentially expressed genes in MDM following infection through the three time points analysed. Furthermore down-regulated genes were only found at 24 h post-infection. Ingenuity Pathways Analysis of differentially expressed genes showed that “cell-mediated immune response” was the most significant network related to 2hpi dataset, “immune cell trafficking” for 6hpi, and “inflammatory response” for 24hpi. Finally the analysis of down-regulated genes at 24hpi confirmed the role of complement and major histocompatibility complex (MHC) in the pathogenesis of MAP in cattle.

Submitted by Cássio da Cruz Nogueira (cassionogueirakk@gmail.com) on 2017-05-05T11:56:57Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_EstudoParatuberculoseBufalos.PDF: 1575243 bytes, checksum: 82baedacd1f3e101d0cc4918e2b8243b (MD5) / Approved for entry into archive by Edisangela Bastos (edisangela@ufpa.br) on 2017-05-10T12:05:55Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_EstudoParatuberculoseBufalos.PDF: 1575243 bytes, checksum: 82baedacd1f3e101d0cc4918e2b8243b (MD5) / Made available in DSpace on 2017-05-10T12:05:55Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_EstudoParatuberculoseBufalos.PDF: 1575243 bytes, checksum: 82baedacd1f3e101d0cc4918e2b8243b (MD5) Previous issue date: 2016-03-11 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / INCT - Instituto Nacional de Ciência e Tecnologia / Este trabalho objetivou estudar a paratuberculose (PTB) em bubalinos no estado do Maranhão (MA). Para isso foram realizadas a utilização da biópsia retal como ferramenta auxiliar na coleta de amostras para o diagnóstico ante mortem da PTB; dosagem de teores de cobre (Cu), zinco (Zn) e ferro (Fe) em animais positivos para PTB, assim como, o relato da detecção de Mycobacterium avium subesp. paratuberculosis (Map) em útero e feto bubalino. No primeiro estudo foram realizadas 140 biopsias retais em bubalinos das raças Murrah, Mediterrâneo e seus mestiços, com idade acima de três anos, no município de São Mateus, MA. Essas biópsias foram processadas por meio das técnicas de Ziehl-Neelsen (ZN) e reação em cadeia da polimerase quantitativa (qPCR). Adicionalmente foi realizada necropsia de 11 búfalos com sinais clínicos sugestivos de PTB, coletadas amostras de íleo e linfonodo mesentério (LM) para a realização de ZN e qPCR. Pela colaboração de ZN das biopsias retais, 4,3% (6/140) apresentaram bacilos álcool-ácidos resistentes (BAAR) e na qPCR, 5% (7/140) tiveram amplificação do material genético. As lesões anatomopatológicas de quatro animais foram compatíveis com lesões sugestivas de PTB, apresentaram BAAR e amplificação de material genético. A concordância entre a biopsia retal e a analise dos tecidos de íleo e LM, segundo o teste Kappa foi alta (k=0,792). No segundo estudo, foram realizadas 13 necropsias em búfalos de raças Murrah, mediterrâneo e seus mestiços, com idade acima de três anos, nos municípios de São Mateus e São Luís, MA. Foram coletadas amostras de íleo e LM para pesquisa de Map e de fígado para dosagem de minerais (Cu, Zn e Fe). Sete búfalos foram positivos PTB e alocados no Grupo 1; e seis búfalos foram negativos e alocados no Grupo 2. Na dosagem dos microminerais, os búfalos do Grupo 1 apresentaram níveis abaixo dos valores de referência para Cu e Zn. observou-se que as medias dos teores de Cu foi 18,0ppm. Os teores de Fe em ambos os grupos foram elevados (>669ppm). O terceiro estudo foi conduzido em uma propriedade em São Luís, MA. Uma búfala prenha, com acentuada debilidade física e com suspeita clinica de PTB foi eutanasiada e necropsiada. Foram coletadas fragmentos de íleo, LM, útero e placenta da fêmea; fragmentos de rim, fígado, vaso umbilical e sistema digestivo (SD) do feto e analisadas através de ZN e qPCR. A fêmea foi positiva em amostras de íleo, LM e útero na qPCR e o feto foi positivo no SD. No íleo, LM e nas fezes foram observados BAAR. Conclui-se que a analise de biopsia retal pode ser empregada para auxiliar no diagnostico ante mortem. A PTB pode induzir quadros de deficiência mineral com consequente piora do quadro clínico da doença. E a presença de Map em búfalos pode ocorrer em vários órgãos, inclusive no sistema reprodutivo e a transmissão intrauterina pode ocorrer nesta espécie. / This study investigated paratuberculosis (PTB) in water buffaloes in the state of Maranhão (MA). For that were performed using the rectal biopsy as an auxiliary tool in the collection of samples for ante mortem diagnosis of PTB; dosages of copper (Cu), zinc (Zn) and iron (Fe) in PTB positive animals was performed, as well as the report of the detection of Mycobacterium avium subsp. paratuberculosis (Map) in the utero and fetus of a buffalo. In the first study were conducted 140 rectal biopsies in buffaloes of Murrah, Mediterranean and their crossbreeds, over the age of three years in São Mateus, MA. These biopsies were processed by Ziehl-Neelsen (ZN) staining and quantitative PCR (qPCR). Postmortem examination of 11 buffaloes with clinical signs suggestive of PTB was performed and samples of ileum and mesenteric lymph nodes (MLN) were collected for performing qPCR and ZN. By ZN staining of rectal biopsies, 4.3% (6/140) acid-fast bacilli and qPCR, 5% (7/140) had amplification of genetic material. The pathological lesions of four animals were consistent with lesions suggestive of PTB showed acid-fast bacilli and amplification of genetic material. The correlation between rectal biopsy and analysis of ileal tissue and LM, according to the Kappa test was high (K=0.792). In the second study, 13 postmortem examination were carried in buffaloes of Murrah, Mediterranean and their crossbreeds, over the age of three years in the cities of São Mateus and São Luís, MA. Ileum and mesenteric lymph nodes samples were collected for Map search and liver to dosage minerals (Cu, Zn and Fe). Seven buffaloes were positive for PTB and allocated in Group 1; six buffaloes were negative and allocated in Group 2. In the dosing of trace minerals in Group 1 showed levels below the reference values for Cu and Zn. It was observed that the average Cu concentration was 18.0ppm and Zn 68.6ppm. In Group 2 the average Cu concentration was 113.7ppm and Zn 110.0ppm. The iron concentrations in both groups were high (>669ppm). The third study was conducted on a property in São Luís, MA. A buffalo cow pregnant with marked physical debility and clinical signs suggestive of PTB was eutanasiada and necropsiada. Fragments ileum, MLN, uterus, and placenta of buffalo cow were collected; fragments of kidney, liver, umbilical vessel and digestive system (DS) of the fetus were collected and were analyzed by Zn and qPCR. The buffalo cow was positive in ileum samples, mesenteric lymph nodes and uterus in qPCR and the fetus was positive in DS. In the ileum, MLN and feces were observed acid-fast bacilli. It is concluded that rectal biopsy analysis can be used to help diagnose ante mortem. The PTB could lead to mineral deficiency and worsen the clinical condition of the buffaloes. The presence of Map in buffalo can occur in various organs, including the reproductive system and intrauterine transmission can occur in this species.