Chapter 1: Introduction and Literature Review; Chapter 2: A retrospective study of anthrax on the Ghaap plateau, Northern Cape Province of South Africa, with special reference to the 2007 - 2008 outbreaks; Chapter 3: Insights gained from sample diagnostics during anthrax outbreaks in the Kruger National Park, South Africa; Chapter 4: Through the lens: a microscopic and molecular evaluation of archival blood smears from the 2010 anthrax outbreaks in Kruger National Park, South Africa; Chapter 5: A distribution snapshot of anthrax in South Africa: multiple locus variable number of tandem repeats analyses of Bacillus anthracis isolates from epizootics spanning 4 decades across southern Africa; Chapter 6: Isolation and Whole Genome Analysis of a Lytic Bacteriophage Infected Bacillus anthracis Isolate from Pafuri, South Africa; Chapter 7: Characterisation of temperate bacteriophages infecting Bacillus cereus sensu stricto group in the anthrax endemic regions of South Africa; Chapter 8: General discussion, conclusions and recommendations. / Thesis (PhD)--University of Pretoria, 2016. / German Research Foundation (DFG) / National Research Foundation (NRF) of South Africa. / Veterinary Tropical Diseases / PhD / Unrestricted

La toxocariasis es una zoonosis de tipo parasitaria producida por un nemátodo del género Toxocara, un parásito zoonótico que infecta a perros y gatos. La infección humana se da al ingerir los huevos larvados presentes en lugares contaminados o directamente en perros o gatos parasitados. Una vez ingerido el huevo larvado por el humano, este migra a diversos órganos del cuerpo, pero no llega a desarrollar la fase adulta. La infección por larvas, dependiendo de la especie de parásito y su ubicación, puede producir: larva migrante cutánea, larva migrante visceral, larva migrante ocular y larva migrante neural. La infección por larvas de Toxocara canis produce larva migrante visceral y ocular. La seroprevalencia humana es alta en los grupos más susceptibles como los niños o los criadores de mascotas. En este sentido, se pensó que los estudiantes de medicina veterinaria podrían estar más expuestos a la infección, de forma particular los de la UMNSM, ya que su facultad cuenta con una clínica veterinaria. Por ello, el presente trabajo buscó determinar la seroprevalencia de toxocariasis en los estudiantes de la Facultad de Medicina Veterinaria de la UNMSM, para lo que se utilizó una prueba de ELISA que emplea el antígeno recombinante Tc-CTL-1 de larvas de Toxocara canis con una sensibilidad de 100% y especificidad de 95.6%. Con este fin se realizó un muestreo aleatorio en estudiantes desde 1er a 6to año. De las 117 muestras de suero obtenidas, dos fueron positivas. Para analizar los resultados se realizó una simulación beta. Los intervalos de confianza del 95% se calcularon empleando la prueba de Bootstrap con 30,000 iteraciones. El intervalo de confianza de la seroprevalencia de toxocariasis en estudiantes de la Facultad de Medicina Veterinaria fue de 0.025 a 0.975. Se concluyó que estudiar la carrera de medicina veterinaria no implica un mayor riesgo de toxocariasis a pesar de la cantidad de años que se está en esta facultad y de contar con una la clínica veterinaria siempre abierta al público. / Tesis

The vectors of avian malaria (Haemosporida) are an understudied component of wildlife disease ecology. Most studies of avian malaria have focused on the secondary bird hosts. This imbalance leaves a significant gap in our knowledge and understanding of the insect hosts. This study investigates the diversity of malaria parasites carried by mosquitoes (Diptera, Culicidae) in the state of Mississippi. Using PCR techniques, haemosporidian infection rates were determined and parasites were identified in a phylogenetic context to those previously annotated. A total of 27,157 female mosquitoes representing 15 species were captured. Five of those species tested positive for malaria parasites with an overall infection rate of 4 per 1000 mosquitoes infected. Mosquitoes were shown to harbor Plasmodium and Haemoproteus parasites. Surprisingly, a unique lineage of parasites was discovered in Anopheles mosquitoes potentially representing a new genus of haemosporidian parasites, reinforcing the need to continue investigating this diverse group of parasites.

Hepatitis E virus (HEV), the causative agent of hepatitis E, is endemic in many developing countries. However, sporadic cases of acute hepatitis E have also been reported in industrialized countries including the United States. Increasing evidence suggested that hepatitis E is zoonotic. Swine HEV was discovered in 1997 from a pig in the United States and has the ability to cross species barrier and infect humans. There are four major genotypes of HEV worldwide and swine HEV identified to date in different countries belongs to either genotypes 3 or 4. Thus far, genotypes 1 (Asian strains) and 2 (a single Mexican strain) of HEV are exclusively found in humans. To determine if genotypes 1 and 2 of HEV also exist in pigs we tested serum and/or fecal samples for from pigs of different age groups in Thailand, and from pigs 2-4 months-of-age in two states (Sonora, Sinaloa) in Mexico. A universal RT-PCR was first standardized to detect all 4 different genotypes of HEV. Swine HEV RNA was detected from in 10/26 pigs at 2-4 months-of-age but not in pigs of 1-, 6-month old, adult/sow pigs from Thailand. In Mexico, swine HEV RNA was detected in 8 of 125 serum samples, 28 of 92 fecal samples of 2-4 month-old pigs. Antibodies to swine HEV were detected in 101 of 125 (80.8%) Mexican pigs. A total of 44 swine HEV isolates were amplified and sequenced for the ORF2 capsid gene region. Sequence analyses revealed that all the swine HEV isolates identified from pigs in Thailand and Mexico belong to genotype 3. Overall, the Mexican swine HEV isolates shared 89-100% sequence identity to each other, and about 89-92% identity with the prototype genotype 3 US swine HEV. The Thailand swine HEV isolates displayed 97-100% nucleotide sequence identity with each other, and 90-91% identity with the prototype genotype 3 swine HEV. Phylogenic analysis revealed that minor branches do exist among Mexican swine HEV isolates. The results from this study indicated that genotype 1 or 2 swine HEV does not exist in pig from countries where human genotypes 1 and 2 HEVs are prevalent. / Master of Science

Submitted by Anderson Silva (avargas@icict.fiocruz.br) on 2012-05-09T12:47:47Z No. of bitstreams: 1 zilene_m_pereira_ioc_ebs_0008_2010.pdf: 15520398 bytes, checksum: 493b6f7c7e7dea5020af412b5cc0cffc (MD5) / Made available in DSpace on 2012-05-09T12:47:47Z (GMT). No. of bitstreams: 1 zilene_m_pereira_ioc_ebs_0008_2010.pdf: 15520398 bytes, checksum: 493b6f7c7e7dea5020af412b5cc0cffc (MD5) Previous issue date: 2010 / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil. / Achatina fulica, ou caramujo africano, foi introduzido no Brasil em meados da década de 1980 para ser comercializado como “escargot”. No entanto, em virtude da baixa demanda do mercado consumidor, os criadores desativaram seus plantéis, gerando fuga ou liberação intencional de exemplares que, livres no ambiente, espalharam-se rapidamente por quase todo país (24 estados mais o Distrito Federal). As “densas” populações do caramujo africano causam uma série de problemas, como a destruição de hortas e jardins, competição com a fauna nativa, além de poderem transmitir zoonoses como a angiostrongilose abdominal e a meningite eosinofílica. A exemplo do que ocorre em outras partes do país, o Município de Barra do Piraí, Estado do Rio de Janeiro, encontra-se também infestado. Visando contribuir para o enfrentamento dos danos causados por essa espécie, o presente estudo objetivou: pesquisar a helmintofauna de populações de A. fulica em alguns bairros de Barra do Piraí quanto à presença de larvas de helmintos de importância médico-veterinária; analisar as percepções relacionadas ao caramujo africano entre estudantes do Ensino Fundamental de uma escola pública estadual no referido município. A pesquisa, de abordagem qualitativa, foi realizada em duas etapas: 1) coleta de exemplares de A. fulica nos bairros Arthur Cataldi, Dorândia, Química, Boca do Mato e São João no período de fevereiro de 2008 a novembro de 2009, os quais foram analisados no Laboratório de Malacologia do Instituto Oswaldo Cruz/Fiocruz (Referência Nacional em Malacologia Médica); 2) aplicação de 82 questionários e a realização de quatro grupos focais com alunos do Ensino Fundamental, além de observações de campo. Os resultados obtidos ao longo do trabalho “comprovam” que há uma infestação de caramujos africanos no município. A pesquisa da helmitofauna revelou que caramujos provenientes de dois dos cinco bairros analisados, quais sejam, Química e Dorândia, apresentaram o nematódeo Angiostrongylus cantonensis causador da meningite eosinofílica no homem. Além disso, também foram encontrados no município os nematódeos Strongyluris-like e Aelurostrongylus abstrusus que causam doenças em animais. Os dados referentes ao estudo das percepções revelaram que os alunos identificam a presença do caramujo africano no contexto no qual estão inseridos, e embora a maioria acredite que esse animal possa causar doenças, apenas uma minoria afirma conhecer pessoas que tiveram problemas com o caramujo. Os dados igualmente apontam para divergências entre a visão dos alunos e o conhecimento científico sobre as formas de controle de A. fulica e os tipos de agravos, em função do contato ou ingestão dessa espécie. Os laudos referentes ao exame dos moluscos e as recomendações aos órgãos municipais sobre o controle e monitoramento A. fulica já foram encaminhados à secretaria de saúde do município para que sejam tomadas as devidas providências. Da mesma forma o trabalho será disponibilizado aos professores da escola que participou desta pesquisa para que suas contribuições possam auxiliar na prática docente. Espera-se que os resultados deste trabalho possam subsidiar o desenvolvimento de ações de controle e monitoramento de A. fulica no município, bem como ações educativas em contextos formais e não formais de ensino para trabalhar com essa temática, adequadas à realidade da comunidade. / The giant African snail Achatina fulica, was introduced in Brazil in mid-1980 for commercial purposes ("escargot" farming) that were not successful. In spite of marketing efforts the demand was low since Brazilians are not used to eat terrestrial snails. Thus, farmers gave up rearing the snails and unaware of the potential negative consequences released the snails by putting them in the garbage, discarding them on waste land and the edges of highways or throwing them into the rivers. The result was a quick spread throughout most of the country (24 states plus the Federal District). The "dense" populations of A. fulica cause many of problems, such as nuisance and destruction of vegetable gardens and gardens. They may also transmit zoonosis such as abdominal angiostrongylosis and meningoencephalitis. Similarly to other municipalities in the state of Rio de Janeiro, Barra do Piraí is also infested. The present study aimed to investigate the larval helminthes of medical and veterinary importance associated with populations of A. fulica in some districts of that municipality as well as analyze the knowledge and perceptions about that snail among students in a public school. Therefore, methodological procedures were performed in two steps: 1) collecting of specimens of A. fulica from five neighborhoods from February 2008 to November 2009, which were analyzed at the Laboratory of Malacology (Instituto Oswaldo Cruz/Fiocruz), a National Center for Medical Malacology Research; 2) application of 82 questionnaires and conducting four focus groups with elementary school students, and field observations. The search for larval helminthes detected the presence of nematodes of medical-veterinary importance: Strongyluris-like, Aelurostrongylus abstrusus and Angiostrongylus cantonensis larvae (causes eosinophilic meningitis in humans) in the neighborhoods of Química and Dorândia. As for the study of the perceptions the data revealed that students identify the presence of the African snail in the context in which they live. Most of the students believe that the snail can cause disease, but a minority said that knew the health problems related to this animal or even people who had them. The data also point to differences between students' views and scientific knowledge on ways to control A. fulica and the types of injuries, depending on contact or ingestion of this species. The reports concerning the examination of snails and recommendations to the municipal authorities over the control and monitoring A. fulica have been forwarded to the secretary of health of the city for taking appropriate action. Moreover, the work will be available to school teachers who participated in this survey for their contributions to assist in teaching. It is hoped that the results of this study may support the development of control measures and monitoring of A. fulica in the county, as well as educational activities appropriate to the reality of the community.

Master of Public Health / Department of Diagnostic Medicine and Pathobiology / M. M. Chengappa / Rabies is the most deadly disease on earth and has a 99.9% human fatality rate. Rabies kills 61,000 humans annually and results in an economic burden of $124 billion USD annually. Each day 3.3 million people live with the risk of rabies. It is estimated that 95% of human rabies cases are a result of coming in contact with an infected canine, majority of these cases being children 15 years and younger. It is estimated that 1 person every 8 minutes dies of rabies. Rabies is a highly neurotropic disease which attacks the brain and central nervous system. Once clinical symptoms are presented, death is invariably the outcome as no cure exists for rabies. Rabies is 100% preventable in humans by proper wound management and proper administration of prophylaxis. Rabies can be adequately controlled in animal populations by contraception and animal rabies vaccine efforts. Whilst it is known that rabies can be prevented in humans and controlled in animal populations, further scientific efforts are still warranted to fully understand this deadly virus so that a cure can one day be discovered. As human and animal populations continue to grow, so does the cost and burden of this horrific disease. As a result, the importance of prophylaxis and passive immunity are critical in the event of medically managing an exposure, and preventing exposures. The World Health Organization has defined global recommendations for individuals and animals who have received prophylaxis to be adequately protected. Measuring this protection is performed using a variety of approved testing methodologies, virus-neutralizing assays and antigen-binding assays. Whilst the WHO recommendations were defined from clinical studies performed with virus-neutralizing assays, the assumption that these recommendations are suitable for the antigen-binding assays is inaccurate. The testing methodologies, virus-neutralization and antigen-binding, share similarities, as they are measuring an immune response to the rabies virus. However; enough differing characteristics are presented such that exact comparisons cannot be made. Establishing the same standards and recommendations for both testing methodologies will never be sufficient.

Memoria para optar al Título Profesional de Médico Veterinario / En el presente estudio se analizaron 153 muestras de heces frescas de gatos domésticos sanos, habitantes de la ciudad de Santiago, Chile, en busca del protozoario zoonótico, Cryptosporidium spp. Las muestras fueron recolectadas en el periodo abril 2009- septiembre 2010, y luego procesadas en el Laboratorio de Parasitología de la Facultad de Ciencias Veterinarias y Pecuarias de la Universidad de Chile, mediante tinción de Ziehl-Neelsen y posterior observación con microscopía óptica, obteniéndose una muestra positiva (0,65%), y cinco sospechosas (3,3%). La muestra positiva pertenecía a una hembra, de 6 meses de edad (grupo etario Kitten), outdoor, de la comuna El Bosque

Orientador: Katia Denise Saraiva Bresciani / Banca: Wagner Luiz Ferreira / Banca: Jancarlo Ferreira Gomes / Resumo: Parasitos gastrintestinais são comuns em animais de companhia. Atualmente, os cães e gatos de estimação apresentam um importante papel entre os humanos, sendo até considerados como membros da família. Por conta deste laço estreito com os humanos, surgem as preocupações dos veterinários em proporcionar uma melhor qualidade de vida para animais, e também em relação a algumas doenças são consideradas de caráter zoonótico. Alguns protozoários e helmintos (Giardia spp., Cryptosporidium spp., Isospora spp., Toxocara spp. e Ancylostoma spp.) são comumente diagnosticados, apesar das medidas terapêuticas e profiláticas existentes. Várias destas enfermidades apresentam sinais clínicos diversos, sendo a diarreia comumente encontrada. As técnicas coproparasitológicas tem como finalidade auxiliar o médico veterinário para um diagnóstico preciso e consequentemente o tratamento adequado ao animal. O objetivo do nosso trabalho foi investigar a ocorrência dos parasitos gastrintestinais em animais filhotes domiciliados. / Abstract: Gastrointestinal parasites are common in pets. Pets play an important role among humans and lately they have been considered members of families Due to the fact of the closely contact between humans and animals, veterinarians worries about zoonotic risk of some parasitic diseases and to promote better life quality for both pets and tutors. Some gastrointestinal parasites (Giardia spp., Cryptosporidium spp., Isospora spp., Toxocara spp. and Ancylostoma spp.) are commonly diagnosed causing diarrhea, despite existing therapeutic and prophylactic evidence. The coproparasitological techniques have been helping veterinarians for an accurate diagnosis and consequently the appropriate treatment to the animal. The objective of this study was to investigate the occurrence of gastrointestinal parasites in domiciled dogs. / Mestre

O Brasil possui o maior rebanho de equinos da América Latina e o terceiro do mundo, gerando um mercado de grande importância econômica para a sociedade brasileira. Esta complexa cadeia de negócios sofre também a influência de variáveis que orbitam as diversas relações estabelecidas. Entre elas, destacam-se as variáveis de ordem macroeconômica (influenciando o mercado e as relações de troca), ambientais e legais, derivadas da ação de órgãos reguladores (MAPA, ANVISA, entre outros). Recentemente, o estado brasileiro se viu refém de uma crise sanitária da zoonose do Mormo em equinos, que afetou diretamente o complexo do agronegócio do cavalo, incluindo dificuldades no âmbito internacional, como a realização das Olimpíadas do Rio 2016. Desta maneira, busca-se contribuir na análise dos pontos críticos ocasionados por esta ameaça de ordem legal/sanitária junto à cadeia de negócios, buscando respostas para melhoria da gestão estratégica entre os atores envolvidos, com base na proposta de gerenciamento de processos de negócios. A abordagem metodológica utilizada nesta pesquisa foi de natureza qualitativa e caracteriza-se como exploratória, quanto ao seu foco, e descritiva, quanto aos seus resultados e conclusões. Na descrição das etapas do Fluxograma de controle e erradicação do Mormo, foram observadas algumas atividades críticas para a consecução deste objetivo no Estado brasileiro. Todas essas atividades foram trabalhadas dentro uma matriz de priorização e, confirmados níveis altos de impacto e de probabilidade, passaram por uma análise de risco. Ao final são formuladas uma série de propostas de melhoria ao Fluxograma atual, buscando-se contribuir na mitigação de riscos para este importante segmento do agronegócio brasileiro. / Brazil has the largest equine population of Latin America and the third one in the world, generating a market of huge economic importance for Brazilian society. This complex business chain also suffers the influence of the variables surrounding these established relationships. Among them, the macroeconomics variables (affecting market and exchange relations), environmental and legal, resulting from the regulatory agencies\" (MAPA, ANVISA and others) actions, are highlighted. Recently the Brazilian state found itself taken into a health crisis of glanders zoonosis in horses, that affected directly the horse agrobusiness complex, including challenges of international context, related to Rio 2016 Olympic Games fulfilment. In this regard, the intention is to contribute towards the analysis of critical points related to this legal/health threat within the business chain, seeking answers for strategic management improvement among engaged actors, based on the propositions of business process management. The research\'s methodologic approach was of qualitative nature; characterized as exploratory regarding their focus, and descriptive in terms of outcomes and conclusions. In the description of the steps of control flowchart and glanders\'s eradication, were sighted some critical activities for the achievement of this goal in Brazil. All of them were worked out within a prioritizin matrix, and confirmed high levels of impact and probability, undergoing a risk analysis. Several improvement proposals for the current flowchart are offered, seeking to contribute towards the mitigation of risks for this fundamental segment of Brazilian agribusiness.

O vírus da hepatite E (HEV) é um patógeno emergente de distribuição global, causador de hepatite aguda e crônica em humanos e infecções assintomáticas em animais. No Brasil a prevalência de infecção por HEV em humanos e animais ainda é pouco compreendida, assim como as características de virulência, patogenicidade e de infecção inter-espécies de isolados do genótipo 3, zoonótico, circulantes no país também são desconhecidas. O estudo foi dividido em duas etapas, com os objetivos de 1) contribuir no diagnóstico laboratorial molecular do HEV a partir do desenvolvimento de um protocolo de RT-PCR em tempo real (RT-qPCR) para pesquisa do HEV em amostras biológicas, e 2) contribuir com a compreensão das características moleculares, sorológicas, clínico-laboratoriais, ultrassonográficas e histopatológicas associadas à infecção experimental em macacos-da-noite (Aotus azarai infulatus) por um isolado do genótipo 3 suíno do HEV previamente detectado na Amazônia oriental brasileira. O protocolo de RT-qPCR foi desenvolvido com a caracterização da curva de detecção e aplicado em concomitância com testes sorológicos para avaliação diagnóstica restrospectiva de 318 (n = 318) amostras de soros humanos suspeitos de hepatite E. O HEV-RNA não foi detectado em nenhuma das amostras humanas testadas, mas foi determinada soroprevalência de 3,4% e 5,9% de anti-HEV IgM e IgG, respectivamente, o que indicou baixa prevalência de infecção por HEV, mesmo entre pacientes com suspeita clínica e/ou laboratorial de hepatite E na Amazônia brasileira. O estudo experimental em macacos-da-noite foi desenvolvido durante 12 semanas e os animais infectados, por via intravenosa (n=3) e oral (n=3) (e dois controles), foram avaliados semanalmente para determinação dos parâmetros clínicos, bioquímicos, hematológicos, sorológicos (pesquisa de anti-HEV IgM e IgG por enzimaimunoensaio) e moleculares (HEV-RNA soro e fezes por RT-qPCR). Adicionalmente, os animais também foram submetidos a avaliação hepática mensal por ultrassonografia, histopatologia e pesquisa hepática de antígenos do HEV por imunohistoquímica. Os seis macacos-da-noite infectados apresentaram o HEV-RNA em amostras de soro e/ou fezes, e alguns apresentaram evidências de soroconversão, detecção hepática do antígeno viral por imunohistoquímica associada a alterações clínicas e laboratoriais de hepatite aguda oligossintomática. Assim, o protocolo RT-qPCR demonstrou ser aplicável na pesquisa molecular do HEV em amostras de humanos e animais, representando uma importante ferramenta de diagnóstico laboratorial. O estudo experimental permitiu a primeira validação de um primata neotropical como modelo experimental para estudos de infecção com o genótipo 3 do HEV. / Hepatitis E virus (HEV) is an emerging pathogen with global distribution that causes acute and chronic hepatitis in humans and asymptomatic infections in animals. In Brazil, the prevalence of HEV infection in humans and animals is still poorly understood, and the characteristics of virulence, pathogenicity and inter-species infection of the genotype 3 isolates circulating in the country are unknown. The study was divided in two stages that aimed to 1) contribute to the molecular diagnosis of HEV infection by the development of a real-time RT-PCR protocol (RT-qPCR) for HEV-RNA research in biological samples, and 2) to contribute to understanding of molecular, serological, clinical-laboratory, ultrasonographic and histopathological features of HEV genotype 3 in owl monkeys (Aotus azari infulatus) experimental infected with isolate of swine HEV genotype 3 previously detected in the Eastern Brazilian Amazon. The RT-qPCR protocol was developed with characterization of a quantification standard curve and later applied concurrently with serological tests in the retrospective evaluation of 318 (n = 318) human serum samples of hepatitis E suspected cases. HEV-RNA was not detected in any of human tested samples, but seroprevalence of 3.4% and 5.9% was determined for anti-HEV IgM and IgG, respectively, that indicated a low prevalence of HEV infection, even among patients with clinical and/or laboratory suspicion of hepatitis E in the Brazilian Amazon. The experimental study in owl monkeys was developed during12 weeks and the animals were infected by intravenous (n = 3) and oral (n = 3) routes (and two negative controls) were evaluated for determination of clinical, biochemical, hematological, serological (anti-HEV IgM and IgG by enzyme immunoassay) and molecular (HEV-RNA serum and stool by RT-qPCR) parameters weekly. Additionally, the animals were also evaluated by hepatic ultrasonography, histopathology and immunohistochemistry research of HEV antigens in liver monthly. The six infected owl monkeys presentend HEV-RNA in serum and/or stool, and some monkeys presented with evidence of seroconversion, liver detection of HEV antigens by immunohistochemistry associated with clinical and/or laboratory findings of oligosymptomatic acute hepatitis. Thus, the RT-qPCR protocol demonstrated to be applicable in the molecular investigation of HEV infection in human and animal samples, and it also represented an important laboratory diagnostic tool. The experimental study allowed the validation of the first neotropical primate model for HEV genotype 3 infection studies.