A vigilância epidemiológica é descrita como o conjunto de procedimentos de coleta, processamento e análise de dados utilizado para dar suporte aos processos de tomadas de decisão por parte dos agentes de saúde animal e humana. A crescente atuação de profissionais do setor privado em sanidade animal tem ampliado a vigilância epidemiológica passiva, com geração de montantes significativos de dados. Neste contexto, surge o Programa Nacional de Controle e Erradicação da Brucelose e Tuberculose Animal - PNCEBT que promove, entre suas estratégias, a habilitação de médicos veterinários privados - MVH - para realização de testes diagnóstico dessas enfermidades em bovídeos. Nesta pesquisa, foi realizada uma compilação de todos os dados gerados pelos MVH relacionados ao PNCEBT no ano de 2008 no Rio Grande do Sul, os quais foram posteriormente analisados com distintas técnicas estatísticas, com objetivo de caracterizar o perfil de ocorrência demográfica e espacial destes testes, bem como buscar indicativos de frequência de enfermidades. Foram analisados resultados de 63.226 testes de bovinos e bubalinos realizados em 5.012 estabelecimentos e 332 municípios do estado, devidamente tratados por procedimentos de imputação múltipla e de detecção de valores extremos. Foi possível verificar que os animais de aptidão leiteira representam a maioria dos animais testados, e que motivação econômica, como bonificações, é o principal estímulo para realização dos testes. Os animais de aptidão leiteira apresentaram, ainda, maior frequência em casos positivos para ambas as enfermidades, quando comparados com as demais aptidões. Os animais do sexo feminino apresentaram maior frequência em ambas as enfermidades, com relação aos machos. Foi verificada maior frequência de animais positivos para tuberculose com o avanço da idade, ao passo que a brucelose se revelou mais freqüente em animais entre 48 e 60 meses. A análise espacial demonstra variações entre regiões nos percentuais de animais testados e de positivos em ambas as enfermidades. As maiores frequências de estabelecimentos positivos para tuberculose ocorreram nas mesorregiões metropolitana e centro-oriental do estado, as quais apresentam altas densidades de estabelecimentos por km². Foi constatado ainda que a distância percorrida pelo MVH para realização de testes pode ser fator limitante na ocorrência dos mesmos, sendo que 49,5% dos testes foram realizados em uma distância máxima de 50 km, considerando-se a ida e o retorno. Foi verificado que municípios onde foram realizados testes têm 67,5 vezes mais chance de ter um MVH residente. Foi possível concluir, assim, que o adequado enfoque aos dados de vigilância permite a realização de inferências relevantes, sendo factível a estruturação de sistemas de monitoramento e vigilância - MOSS. Tais sistemas podem incrementar a qualidade e acessibilidade dos dados, permitindo aos gestores públicos e privados a realização de análises customizadas que tragam maior eficiência e eficácia às ações em saúde animal. / Epidemiological surveillance is described as a set of procedures of collecting, processing and data analysis used for supporting the decision making process of animal and human health managers. The growing participation of private sector professionals on animal health issues has been widening the passive epidemiological surveillance, generating significant amounts of data. In this context, the National Program of Animal Brucellosis and Tuberculosis Control and Eradication – PNCEBT, promotes, amongst other strategies, the accreditation of private veterinarians to make diagnosis tests on bovine, increased significantly the amount of data on bovine tuberculosis and brucellosis. In the present research, a compilation of all data from diagnosis tests performed by accredited private veterinarians on behalf of PNCEBT in the state of Rio Grande do Sul in 2008 was made, which was analyzed through distinct statistics techniques, aiming at characterizing the demography and spatial patterns of occurrence, as well as to assess the frequencies of those diseases. Data from 63.226 individual tests performed in 5.012 farms on 332 municipalities were analyzed, after treatment by multiple imputation and outlier values diagnosis. It was possible to conclude that the majority of tests were from dairy related animals; economical reasons, such as bonus programs, were the main motivation for the farmers to perform tests on their properties. Higher frequencies for both diseases were found in dairy cattle, when compared to other purposes animals. Female presented higher frequencies for both diseases when compared to males. It was also observed that the frequency of tuberculosis increased with age, while for brucellosis the frequency was higher between 48 to 60 months-old animals. Spatial analysis evidenced higher frequencies of positive tuberculosis farms in the metropolitana and centro-oriental mesorregions, which had higher farm density per km². It was also verified that the distance by MVH to perform tests might be a limiting factor for them to occur, being 49.5% of the tests performed at maximum round-way distance of 50 km. The municipalities which had tests performed showed 67.5 times more chances of having a resident MVH. It was possible to conclude that an appropriate approach of surveillance data allows the making of relevant inferences, being feasible the establishment of monitoring and surveillance systems - MOSS. These systems could improve data quality and accessibility, allowing public and private managers to perform custom analysis to bring more efficiency and efficacy to animal health programs.

Considerando que os meios de cultura e as condições de incubação são os principais fatores para o sucesso do primo isolamento, além do método de descontaminação, quatro meios de cultura e três condições de incubação foram investigados. Noventa e sete amostras de lesões granulomatosas foram submetidas ao método de descontaminação com cloreto de 1-hexadecilpiridinio (HPC) a 1,5%, e semeadas em dois meios a base de ovo, Stonebrink e Löwenstein-Jensen com piruvato de sódio, e em dois meios a base de ágar, B83 e Middlebrook 7H11. Cada meio foi incubado a 37ºC por 90 dias em três condições de incubação, atmosfera com 10% de CO2, atmosfera normal e atmosfera com suposta tensão de CO2 obtida pela queima do algodão hidrófobo e fechamento do tubo com rolha de cortiça. O tipo de condição de incubação utilizado teve influência nos meios a base de ovo apenas no inicio da incubação (30 dias), mas nenhuma nos meios a base de ágar. A incubação em atmosfera com 10% de CO2 diminuiu o tempo de aparecimento da primeira colônia e aumentou o número de UFC. O meio B83 foi mais rápido no aparecimento das colônias e teve o maior sucesso de isolamento aos 30 dias, mas não houve diferença com os meios Stonebrink e Löwenstein-Jensen com piruvato, no sucesso de isolamento e número de UFC aos 60 e 90 dias. De acordo com os dados, em sete oportunidades houve isolamento de M. bovis apenas no meio de Stonebrink e em quatro apenas no B83, assim, sugere-se a utilização desses dois meios de cultura em paralelo, incubados em atmosfera com acréscimo de CO2 / Considering that the culture media and the incubation conditions are the main factors for the success of primary isolation, besides the decontamination procedure, four culture media and three incubation conditions were investigated. Ninety-seven samples of granulommatous lesions were submitted to the decontamination procedure by 1-hexadecylpyridinium chloride at 1,5% w/v, and inoculated on two egg-based media, Stonebrink and Löwenstein-Jensen with sodium pyruvate, and two agar-based media, B83 and Middlebrook 7H11. Each medium was incubated at 37ºC for 90 days in three incubation conditions, in air containing 10% CO2, in air, and in air with a supposed higher CO2 tension created by burning the hydrophobic cotton used to close the tubes and subsequently closing with a cork. The type of incubation condition used had influence on the egg-based media only at the beginning of incubation (30 days), but none on the agar-based media. The incubation in air containing 10% CO2 decreased the time to first appearance of colonies and increased the number of colonies. B83 medium showed a faster growth and detected more isolates at 30 days of incubation. However, there was no difference between B83, Stonebrink and Löwenstein-Jensen with pyruvate at 60 and 90 days of incubation. According to the data, seven M. bovis isolates grew only on Stonebrink and four only on B83, therefore, the use of both media, in parallel, incubated in air containing CO2 is suggested.

A vigilância epidemiológica é descrita como o conjunto de procedimentos de coleta, processamento e análise de dados utilizado para dar suporte aos processos de tomadas de decisão por parte dos agentes de saúde animal e humana. A crescente atuação de profissionais do setor privado em sanidade animal tem ampliado a vigilância epidemiológica passiva, com geração de montantes significativos de dados. Neste contexto, surge o Programa Nacional de Controle e Erradicação da Brucelose e Tuberculose Animal - PNCEBT que promove, entre suas estratégias, a habilitação de médicos veterinários privados - MVH - para realização de testes diagnóstico dessas enfermidades em bovídeos. Nesta pesquisa, foi realizada uma compilação de todos os dados gerados pelos MVH relacionados ao PNCEBT no ano de 2008 no Rio Grande do Sul, os quais foram posteriormente analisados com distintas técnicas estatísticas, com objetivo de caracterizar o perfil de ocorrência demográfica e espacial destes testes, bem como buscar indicativos de frequência de enfermidades. Foram analisados resultados de 63.226 testes de bovinos e bubalinos realizados em 5.012 estabelecimentos e 332 municípios do estado, devidamente tratados por procedimentos de imputação múltipla e de detecção de valores extremos. Foi possível verificar que os animais de aptidão leiteira representam a maioria dos animais testados, e que motivação econômica, como bonificações, é o principal estímulo para realização dos testes. Os animais de aptidão leiteira apresentaram, ainda, maior frequência em casos positivos para ambas as enfermidades, quando comparados com as demais aptidões. Os animais do sexo feminino apresentaram maior frequência em ambas as enfermidades, com relação aos machos. Foi verificada maior frequência de animais positivos para tuberculose com o avanço da idade, ao passo que a brucelose se revelou mais freqüente em animais entre 48 e 60 meses. A análise espacial demonstra variações entre regiões nos percentuais de animais testados e de positivos em ambas as enfermidades. As maiores frequências de estabelecimentos positivos para tuberculose ocorreram nas mesorregiões metropolitana e centro-oriental do estado, as quais apresentam altas densidades de estabelecimentos por km². Foi constatado ainda que a distância percorrida pelo MVH para realização de testes pode ser fator limitante na ocorrência dos mesmos, sendo que 49,5% dos testes foram realizados em uma distância máxima de 50 km, considerando-se a ida e o retorno. Foi verificado que municípios onde foram realizados testes têm 67,5 vezes mais chance de ter um MVH residente. Foi possível concluir, assim, que o adequado enfoque aos dados de vigilância permite a realização de inferências relevantes, sendo factível a estruturação de sistemas de monitoramento e vigilância - MOSS. Tais sistemas podem incrementar a qualidade e acessibilidade dos dados, permitindo aos gestores públicos e privados a realização de análises customizadas que tragam maior eficiência e eficácia às ações em saúde animal. / Epidemiological surveillance is described as a set of procedures of collecting, processing and data analysis used for supporting the decision making process of animal and human health managers. The growing participation of private sector professionals on animal health issues has been widening the passive epidemiological surveillance, generating significant amounts of data. In this context, the National Program of Animal Brucellosis and Tuberculosis Control and Eradication – PNCEBT, promotes, amongst other strategies, the accreditation of private veterinarians to make diagnosis tests on bovine, increased significantly the amount of data on bovine tuberculosis and brucellosis. In the present research, a compilation of all data from diagnosis tests performed by accredited private veterinarians on behalf of PNCEBT in the state of Rio Grande do Sul in 2008 was made, which was analyzed through distinct statistics techniques, aiming at characterizing the demography and spatial patterns of occurrence, as well as to assess the frequencies of those diseases. Data from 63.226 individual tests performed in 5.012 farms on 332 municipalities were analyzed, after treatment by multiple imputation and outlier values diagnosis. It was possible to conclude that the majority of tests were from dairy related animals; economical reasons, such as bonus programs, were the main motivation for the farmers to perform tests on their properties. Higher frequencies for both diseases were found in dairy cattle, when compared to other purposes animals. Female presented higher frequencies for both diseases when compared to males. It was also observed that the frequency of tuberculosis increased with age, while for brucellosis the frequency was higher between 48 to 60 months-old animals. Spatial analysis evidenced higher frequencies of positive tuberculosis farms in the metropolitana and centro-oriental mesorregions, which had higher farm density per km². It was also verified that the distance by MVH to perform tests might be a limiting factor for them to occur, being 49.5% of the tests performed at maximum round-way distance of 50 km. The municipalities which had tests performed showed 67.5 times more chances of having a resident MVH. It was possible to conclude that an appropriate approach of surveillance data allows the making of relevant inferences, being feasible the establishment of monitoring and surveillance systems - MOSS. These systems could improve data quality and accessibility, allowing public and private managers to perform custom analysis to bring more efficiency and efficacy to animal health programs.

Made available in DSpace on 2014-07-29T15:07:34Z (GMT). No. of bitstreams: 1 Dis_Cinthya M F Fleury.pdf: 340462 bytes, checksum: 8bfbe121280bc66c1b0b439872531e30 (MD5) Previous issue date: 2010-02-08 / The purpose of this work is to contribute to the research in new diagnostic tests for bovine tuberculosis. To this end a study was performed to identify Mycobacterium bovis immunogenic proteins isolated from bovines in the field. Total extracted protein, were initially fractioned by electrophoresis in a poliacrylamide gel and then identified by Western Blotting, using serum from reagent and non-reagent animals to the purified protein derivate (PPD) intradermal diagnostic test. Protein immunogenicity was assessed according to the molecular weight profile of the positive bands. 93 serum samples were tested from PPD positive and PPD negative animals from Goiás, Distrito Federal and Minas Gerais, as well as tuberculosis free regions of the USA. By means of the Western Blotting technique it was possible to identify the 26kDa protein as the most immunogenic among the positive PPD tested animals. / Com o objetivo de contribuir com pesquisas em novos testes de diagnóstico para a tuberculose bovina, foi realizado um estudo baseado na identificação de proteínas imunogênicas de Mycobacterium bovis isolado de bovinos infectados à campo. Para isso, proteínas totais extraídas de M. bovis caracterizado geneticamente por Spoligotyping, foram inicialmente fracionados por eletroforese em gel de poliacrilamida e posteriormente identificados por Western Blot utilizando soro de animais reagentes e não reagentes ao teste de diagnóstico intradérmico com derivado protéico purificado (PPD). A imunogenicidade das proteínas foi avaliada conforme o perfil do peso molecular das bandas expressadas. Foram testadas um total de 93 amostras de soro de animais PPD positivos e negativos provenientes do estado de Goiás, Distrito Federal, Minas Gerais e região livre de tuberculose dos Estados Unidos. Pela técnica de Western Blot pode-se identificar a proteína de 26kDa como a mais imunogênica dentre os animais PPD-positivos testados.

Bien que la France soit reconnue indemne de tuberculose bovine (TB) par l’Union Européenne depuis 2001, une centaine de nouveaux foyers sont déclarés chaque année. Deux des principaux mécanismes de propagation de l’infection entre troupeaux bovins sont l’introduction d’animaux infectés et les contacts au pâturage. L’objectif de cette thèse a été de quantifier les rôles respectifs des échanges commerciaux de bovins et des contacts au pâturage « par-dessus la clôture » dans la circulation de la TB en France entre 2005 et 2014. Pour cela, nous avons combiné des méthodes d’analyse de réseaux sociaux et de modélisation dynamique. Nos analyses ont montré que les contacts par le commerce étaient une origine probable pour 12% [5 - 18] des infections déclarées en France entre 2005 et 2014, contre 73% [68 - 78] pour la transmission locale. Cependant, nous avons pu observer que les contacts par le commerce permettaient à l’infection de coloniser de nouvelles zones. Puis nous nous sommes intéressés au risque structurel du réseau du pâturage. À partir d’une exploitation donnée, il était possible d’atteindre quasiment toutes les autres exploitations du territoire métropolitain. Il semble donc facile pour l’agent de la TB de se propager par les contacts de voisinage au pâturage. Enfin, afin de reconstituer la propagation de la TB en France entre 2005 et 2014, nous avons développé un modèle dynamique stochastique individu-centré, dirigé par les données et avec un pas de temps mensuel. Ce modèle reproduit une dynamique intra- et inter-élevages de l’infection, avec une transmission de l’infection entre élevages par le commerce et au pâturage. Il a permis de confirmer la facilité de propagation de l’infection via les contacts au pâturage. En conclusion, les mécanismes de propagation de la TB sont complémentaires : le commerce permet une diffusion sur de longues distances et le pâturage facilite la propagation locale. Le dispositif de lutte contre la tuberculose bovine doit donc s’appuyer combiner les mesures de contrôle afin de cibler plusieurs mécanismes de propagation. / Although France was declared free of bovine Tuberculosis (bTB) by the European Union in 2001, a hundred of outbreaks are reported every year. Cattle movements and contacts on pastures are considered the main between-herd transmission routes of bTB. The aim of this work was to quantify the respective roles of live cattle trade and “over the fence” contacts on pastures in the spread of bTB in France between 2005 and 2014, using social network analysis and dynamic modelling. Results showed that cattle movements may be the origin of 12% [5 - 18] of the French outbreaks between 2005 and 2014, versus 73% [68 - 78] due to the transmission on pasture. However, cattle movements appeared to allow the introduction of the bacteria into new areas. In a second step, the structural risk resulting from the French pasture network was studied. Almost all of the premises owning pastures were mutually connected by pastures neighbourhood relationships, on the territory. Thus, the spatial configuration of pastures appeared favourable to the spread of bTB in France. Finally, to mimic the spread of the infection in France between 2005 and 2014, a stochastic individual-based model was developed. It was driven by the data with a monthly time step. This model reproduced the within- and between-herd dynamic, with a between-herd transmission due to cattle movements and contacts on pastures. The easy spread of the bacteria through the network of French pastures was confirmed using the model. In conclusion, cattle movements and contacts on pastures are complementary paths for bTB spread: cattle movements allow the long-range spread, whereas contacts on pasture make easier the short-range spread. Hence, control programs must target several routes of transmission to be able to eradicate bTB.

La tuberculose bovine est une maladie zoonotique se transmettant principalement par inhalation. C’est une maladie qui peut avoir des répercussions économiques (ex : saisie d’un animal infecté à l’abattoir) et sur les échanges commerciaux, car les pays peuvent bannir l’importation des animaux provenant des pays touchés par cette maladie. Au Canada, le contrôle de cette maladie a commencé en 1923 chez les bovins et a ensuite été appliqué chez les cervidés depuis 1989. Ce programme est composé par la surveillance, la réponse aux éclosions, ainsi que le contrôle des mouvements des cervidés. La capacité de détecter cette maladie lors des activités de surveillance est influencée par la performance des méthodes diagnostiques utilisées à chaque étape de la surveillance. À l’abattoir, la surveillance est effectuée par l’inspection visuelle post-mortem et par différentes procédures diagnostiques réalisées sur des prélèvements de tissus anormaux (histopathologie, réaction de polymérisation en chaîne (PCR) ou culture bactérienne). Chez le cervidé vivant, la surveillance est plutôt réalisée par des tests cutanés ou sérologiques. L’objectif principal de ce projet était de fournir, à travers une revue systématique de littérature, les estimés des valeurs des principaux paramètres pouvant influencer la probabilité de détection de la tuberculose bovine dans un contexte de surveillance de cette maladie à l’abattoir chez les cervidés d’élevage et chez les cervidés vivants. Les études qui ont estimé la probabilité de présence des lésions macroscopiques ont trouvé qu’elle variait de 20% à 100% alors que la probabilité de détection des lésions tuberculeuses était supérieure ou égale à 90%. L’estimé de la sensibilité des tests diagnostiques pouvant être utilisés pour la détection de la tuberculose bovine à l’abattoir (histopathologie, PCR et culture) variait de 63.6% à 98.6%. Chez les animaux vivants, les tests sérologiques et sanguins avaient une sensibilité qui variait de faible à élevée (1.4% à 100.0%) alors que les tests cutanés avaient une sensibilité qui variait de 65.0% à 100.0%. Comme le stade de la maladie (temps depuis l’infection) influence la sensibilité de la majorité des tests diagnostiques et que ce facteur n’a généralement pas été rapporté par les études, la performance de ces tests peut difficilement être comparée. La prévalence de la tuberculose estimée lors des études transversales ou de contrôle de la maladie était faible (prévalence au niveau animal au sein d’un troupeau). Une étude effectuée au Canada sur les facteurs de risque de la tuberculose bovine chez les cervidés d’élevage a constaté que l’augmentation de la taille du troupeau était associée à une augmentation du risque de positivité (du troupeau) à la tuberculose bovine. Ces informations sur la performance et les limites des méthodes diagnostiques sont essentielles pour orienter le choix du test diagnostique et pour permettre l’évaluation de la sensibilité des activités de surveillance / Bovine tuberculosis is a zoonotic disease mostly transmitted by inhalation of contaminated material. This disease leads to the economic losses (e.g. condemnation of meat from infected animals at the abattoir) and impede trade since countries can stop importing live cervids from tuberculosis endemic countries. In Canada, the control of bovine tuberculosis started in 1923 in cattle and then was implemented in farmed cervids in 1989. This program is composed of surveillance, response to the outbreaks and movement control of cervids. The capacity of detecting this disease during the surveillance activities depends on the performance of diagnostic methods used at each step of the component of the surveillance system. At the abattoir, the inspection is carried out by visual inspection of organs and carcass and diagnostic tests carried on abnormal tissues by using histopathology, PCR or culture. In live cervids, the detection is carried out through skin or serological testing. The main objective of this project was to provide, through a systematic literature review, the estimates of main parameters that can influence the probability of detecting bovine tuberculosis in the context of surveillance activities in farmed cervids at the abattoir and in live cervids. The probability of presence of macroscopic lesions estimated by studies ranged from 20% to 100% while the estimate of probability of detection of lesions was greater than or equal to 90%. The estimate of sensitivity of diagnostic tests used during bovine tuberculosis surveillance at the abattoir (histopathology, PCR and culture) ranged from 63.6% to 98.6%. In live cervids, studies that investigated the performance of serological tests reported an estimate of sensitivity ranging from low to high (1.4% to 100.0%) while skin tests were reported to have a sensitivity ranging from 65.0% à 100.0%. Given that the stage of the disease affects the performance of most diagnostic test while that variable was not reported by studies, it was difficult to compare the performance of the various tests. Bovine tuberculosis prevalence estimated by studies using a cross-sectional or disease control design reported a low prevalence (prevalence at the animal level within a herd). One study carried out in Canada evaluated the risk factors for bovine tuberculosis infection and found that the increase of herd size was associated with an increase of risk of herd positivity to bovine tuberculosis. This information on the performance and limits of bovine tuberculosis diagnostic tests is critical for orienting the choice of appropriate diagnostic method and for a valid evaluation of the surveillance activities

A tuberculose é uma das principais preocupações da Organização Mundial da Saúde, principalmente após o surgimento da AIDS que alavancou os índices da doença, sendo considerada a principal causa de morte por um único agente. Além do M. tuberculosis, agente responsável pela doença em humanos, outra manifestação importante é a tuberculose em bovinos causada pelo M. bovis, que também apresenta importância epidemiológica, devido à transmissão para o homem pela ingestão de alimentos contaminados, e a escassez de dados com relação à sua prevalência na população. Programas de controle da doença nos rebanhos bovinos existem em todo o mundo, e os maiores índices da doença encontram-se nos países em desenvolvimento. Estes programas estão baseados na identificação dos animais positivos, por meio de testes de tuberculina e sacrifício dos mesmos. Lesões encontradas em exames post-mortem podem ser submetidas a estudos bacteriológicos para o isolamento e identificação do agente e histopatológicos para a caracterização da lesão, os quais demandam meses para a sua conclusão. Com o advento da biologia molecular, novos métodos são propostos para a diminuir o tempo do diagnóstico de meses para poucos dias. Com o intuito de proporcionar um panorama das novas metodologias moleculares, como a técnica de PCR, empregadas no diagnóstico da tuberculose bovina, realizou-se uma revisão bibliográfica, apresentando as vantagens e dificuldades das mesmas. Apesar dos avanços alcançados com estas técnicas, a padronização de métodos viáveis para a rotina de diagnóstico laboratorial ainda não foi obtida, sendo essencial o investimento em pesquisas com o objetivo de solucionar estas barreiras. / Tuberculosis is one of the main concern of World Health Organization, especially after the appearance of the AIDS that increased the rate of this disease that is the principal cause of death by one unique agent. Besides the tuberculosis caused by M. tuberculosis in human being, the disease caused by M. bovis in man shows epidemiological importance by its transmission trough contaminated food and the requirement of datas about its prevalence in human being. Control programs of bovine tuberculosis are present worldwide and the major rates of the disease are found in developing countries. This programs are based on test-and-slaughter, defined by the application of tuberculin test to cattle and slaughter of positive animals. Tuberculous lesions founded at post mortem inspection can be analyzed by bacteriological methods for isolation and identifying of the agent and histopathological examination that both require several days to conclusion. After the advent of molecular biology, new methods have been proposed to reduce the time of diagnostic from moths to few days. With the meaning to provide an overview of the new molecular methods applied to the bovine tuberculosis diagnosis, as PCR method, a review as carried out showing those advantages and difficulties. Although the advances reached by these techniques, the standardization of viable methods to the routine laboratory diagnosis couldn?t be reached and the investment in researches is essential to solve these barriers.

This thesis presents the results of a series of studies on the epidemiology of TB in brushtail possum and domestic cattle populations in New Zealand. The first set of studies provides an analysis of the results of routine TB testing carried out in the Featherston area from July 1980 to June 2004. The median annual incidence rate of TB reduced from 4.7 cases per 1000 cattle-years at risk for the period 1986 to 1991 to 1.8 cases per 1000 cattle-years at risk for the period 1992 to 2003, coincident with the use of poisoning to control possums in the surrounding forest park (a major possum habitat area). We identified clusters of cattle TB cases adjacent to the forest park and found no evidence of spatio-temporal interaction of TB risk among farms. Our findings support the hypothesis that possums living in the forest park are a source of bovine TB in this area and that farm-to-farm transmission was not an important mechanism of infection spread. A mixed-effects Poisson regression model was developed to investigate the influence of farm-level covariates on the number of cattle confirmed with TB. The model showed that, despite intensification of possum control activities, proximity to forest parks remained a significant predictor of the number of confirmed TB cases per farm per year. Our analyses identified a significant, 3-fold increase in TB risk in dairy cattle relative to beef conditional on the size of local possum habitat, and confirmed the positive influence of cattle population size and the presence of previous infection status as a determinant of the number of confirmed TB cases per farm per year. The second set of studies investigates details of capture events recorded during a longitudinal, capturemark- recapture study of possums in a 22-hectare study site near Castlepoint, from April 1989 to August 1994. Social network analyses were used to identify contact patterns and to estimate the influence of contact on R0 for bovine TB. The average number of contacts per possum ranged from 20 to 26 per year. We estimated that TB would spread if an average of between 1.94 and 1.97 infective contacts occured per year per infected possum. We evaluated the effect of sex, habitat and contact behaviour of 26 postmortem confirmed TB cases in possums with those of 104 matched controls. Unit increases in the number of infected contacts increased the odds of TB infection by 2.61 (95% CI 1.29 – 5.29, P <0.01). Our results show that individual contact behaviour is a determinant of the presence of TB foci within this population and challenge the hypothesis that contact with many individuals increases the probability of infection. A model to predict spatial variation in possum abundance was developed using a Geographic Information System. Details of possum capture events were obtained from 157 10-trap lines distributed within 42 randomly located transects at Molesworth Station. Two GIS-based models were developed to predict the number of possums caught per line using Poisson regression techniques. The first model used remotely sensed environmental data; the second used a combination of remotely sensed and fine-scale data. Both models provided adequate predictive ability with Pearson correlation coefficients greater than 60%. We conclude that the prediction maps produced from this model provide a useful decision support tool for possum control managers. These results have implications for the management of TB in this area of New Zealand, providing the information that will allow effective control activities to be applied at significantly lower cost.

Diseases that infect wildlife populations pose a significant threat to public health, agriculture, and conservation efforts. The spread of these diseases can be influenced by the social structure of the population, and therefore often need to be accounted for in disease models. In this thesis I use high-resolution contact data to explore the social structure of a high-density population of European badgers (Meles meles). I explore how this structure might influence the spread of bovine tuberculosis (bTB), a debilitating disease of cattle for which badgers are a wildlife reservoir. Denning and home range data collected using radio tracking is also used to determine how this social structure is related to badger space use. I use social network analysis to identify the community structure of the badger population, revealing that badgers interact in fewer, more distinct groups than previously assumed. This is likely to inhibit the spread of disease through the population, given that the probability of infection entering a new social group will be reduced. However, among-group contact is still found to occur even between the most isolated groups. I show that this among-group contact is more likely to occur between less related individuals, possibly suggesting that breeding behaviour may drive among-group contact as a mechanism for inbreeding avoidance. To gain additional insight into this among-group contact, I determine how badger spatial behaviours are related. I show that the use of dens (setts) away from the social group’s main sett (outlier setts) in the spring is associated with extra-territorial ranging. I also show that this extra-territorial ranging is associated with more central network positions. The seasonality of this behaviour further suggests that this may be related to breeding activity. These findings suggest that behaviours associated with extra-group ranging may increase the risk of acquiring and transmitting infection. Therefore, use of outlier setts in the spring could act as a spatial proxy to identify high-risk individuals for disease spread, offering potential targets for disease control. Finally, I discuss the implications of these findings in regard to what they reveal about badger behaviour, disease transmission, and the design of effective disease control strategies. The importance of understanding population social structure for the study of wildlife disease in general is also discussed.

A tuberculose é uma das principais preocupações da Organização Mundial da Saúde, principalmente após o surgimento da AIDS que alavancou os índices da doença, sendo considerada a principal causa de morte por um único agente. Além do M. tuberculosis, agente responsável pela doença em humanos, outra manifestação importante é a tuberculose em bovinos causada pelo M. bovis, que também apresenta importância epidemiológica, devido à transmissão para o homem pela ingestão de alimentos contaminados, e a escassez de dados com relação à sua prevalência na população. Programas de controle da doença nos rebanhos bovinos existem em todo o mundo, e os maiores índices da doença encontram-se nos países em desenvolvimento. Estes programas estão baseados na identificação dos animais positivos, por meio de testes de tuberculina e sacrifício dos mesmos. Lesões encontradas em exames post-mortem podem ser submetidas a estudos bacteriológicos para o isolamento e identificação do agente e histopatológicos para a caracterização da lesão, os quais demandam meses para a sua conclusão. Com o advento da biologia molecular, novos métodos são propostos para a diminuir o tempo do diagnóstico de meses para poucos dias. Com o intuito de proporcionar um panorama das novas metodologias moleculares, como a técnica de PCR, empregadas no diagnóstico da tuberculose bovina, realizou-se uma revisão bibliográfica, apresentando as vantagens e dificuldades das mesmas. Apesar dos avanços alcançados com estas técnicas, a padronização de métodos viáveis para a rotina de diagnóstico laboratorial ainda não foi obtida, sendo essencial o investimento em pesquisas com o objetivo de solucionar estas barreiras. / Tuberculosis is one of the main concern of World Health Organization, especially after the appearance of the AIDS that increased the rate of this disease that is the principal cause of death by one unique agent. Besides the tuberculosis caused by M. tuberculosis in human being, the disease caused by M. bovis in man shows epidemiological importance by its transmission trough contaminated food and the requirement of datas about its prevalence in human being. Control programs of bovine tuberculosis are present worldwide and the major rates of the disease are found in developing countries. This programs are based on test-and-slaughter, defined by the application of tuberculin test to cattle and slaughter of positive animals. Tuberculous lesions founded at post mortem inspection can be analyzed by bacteriological methods for isolation and identifying of the agent and histopathological examination that both require several days to conclusion. After the advent of molecular biology, new methods have been proposed to reduce the time of diagnostic from moths to few days. With the meaning to provide an overview of the new molecular methods applied to the bovine tuberculosis diagnosis, as PCR method, a review as carried out showing those advantages and difficulties. Although the advances reached by these techniques, the standardization of viable methods to the routine laboratory diagnosis couldn?t be reached and the investment in researches is essential to solve these barriers.