Made available in DSpace on 2014-08-20T13:32:51Z (GMT). No. of bitstreams: 1 dissertacao_elizandra_schoenardie.pdf: 268602 bytes, checksum: 08408f44b3e993faf7cf221741aa401e (MD5) Previous issue date: 2005-06-24 / The Visceral Larva Migrans (VLM) is a zoonotic disease caused by the helminth Toxocara cannis. The precocious diagnosis of this disease in humans is very important to determinate the evolution of the clinical case and the patient's treatment. The goal of the first experiment was determinate the presence of antibodies anti-T. cannis in children from Pelotas through the Indirect Enzyme-Linked Immunosorbent Assay (ELISA) front to the antigen TES, as well as to define the pattern of bands recognized by the positive serums in ELISA through the "Western blotting . For this experiment 427 serums from children, with ages between one to eleven yeas old was tested, those serums was adsorved with AgSoAl and determinate that 50,6% was positive for antibodies anti-TES, showing a significant association among the positive children for antibodies anti-TES and the contact with dogs and cats. This association was also observed in the children's different age groups, but not regarding the gender of the same ones. To perform the Western Blotting , 70 serums witch give a positive result in the Indirect ELISA was been used, all serums recognize glicoproteic bands in the range between 30 and 120 kDa. Was observed a diminution in the crusade reaction with AgSoAl when the adsorved and not adsorved serums with this antigen has been test in the Western blotting , where a band of 30 kDa demonstrate to be an important glicoprotein to specific diagnosis of VLM. In the second experiment, 25 mice BALB/c were inoculated with approximately 1000 eggs containing the larvae L3. Each fifteen days blood collection was made through the reto orbital plexus until the 105 days after de animals infection. The serums was tested in the Indirect ELISA using the Antigen TES and urea 6M in order to discriminate recent and late infection, through the percentile of avidity of the IgG in the different days after the infection. A low percentile of avidity was observed to the 15 days after inoculation (between 7,25 and 27,5%). After 60 days of infection, all the animals presented avidity between 31,4 and 58%. This result suggests that in mice BALB/c, to the 60 days after infection the chronic phase of VLM is already established. / A Larva Migrans Visceral (LMV) é uma doença zoonótica que possui como principal agente etiológico o helminto Toxocara canis. O diagnóstico precoce da doença no homem é importante para estudos de evolução clínica e tratamento do paciente e os inquéritos epidemiológicos para determinar a freqüência da infecção em uma população. Por isso, o primeiro experimento teve como objetivo, determinar a presença de anticorpos anti-T. canis em crianças da região de Pelotas através de Enzyme linked immnosorbent assay (ELISA) com o antígeno de excreção e secreção de larvas de Toxocara canis (TES), bem como definir, através de Western blotting , o padrão de bandas do TES reconhecidas pelos soros positivos ao ELISA. Foram ensaiados no ELISA Indireto 427 soros de crianças de um a 12 anos de idade adsorvidos com antígeno somático de Ascaris lumbricoides e determinado que 50,6% apresentaram anticorpos anti-TES, ocorrendo uma associação significativa entre as crianças positivas e o contato com cães e gatos. Esta associação também foi observada em diferentes faixas etárias das crianças, mas não com relação ao sexo das mesmas. Setenta soros positivos no ELISA foram ensaiados no Western blotting e todos reconheceram frações proteicas entre 30 e 120 kDa. Uma diminuição da reação cruzada com o AgSoAl foi observada quando soros adsorvidos com este antígeno foram testados no Western blotting , sendo que uma fração antigênica de 30 kDa apresentou-se como uma proteína importante para o diagnóstico específico da LMV. No segundo experimento, 25 camundongos BALB/c foram inoculados com aproximadamente 1000 ovos contendo a larva infectante (L3). Colheitas quinzenais de sangue foram realizadas através do plexo retro orbital até os 105 dias pós-infecção dos animais. Os soros foram ensaiados no ELISA Indireto utilizando o antígeno TES e a uréia 6M a fim de discriminar infecção recente e tardia, através do percentual de avidez da IgG nos diferentes dias após a infecção. Um baixo percentual de avidez, característico da infecção aguda, foi observado aos 15 dias pós-inoculação (entre 7,3 e 27,5%). Após 60 dias de infecção, todos os animais apresentaram avidez entre 31,4 e 58%. Através destes resultados, sugere-se que em camundongos BALB/c, aos 60 dias pós-infecção a fase crônica da LMV já está estabelecida.

Zoonotic diseases are diseases that are transmitted from animals or vectors to humans and vice versa. The public together with veterinarian authorities should readily access disease information as it is vital in rapidly controlling resultant zoonotic outbreak threats through improved awareness. Currently, the reporting of disease information in South Africa is predominantly limited to traditional methods of Information Communication Technologies (ICTs) like faxes, monthly newspaper reports, radios, phones and televisions. Although these are effective ways of communication, their disadvantage is that the information that most of them offer can only be accessed at specific times during a crisis. New technologies like the internet have become the most efficient way of distributing information in near-real-time. Many developed countries have used web-based reporting platforms to deliver timely information through temporal and geographic visualisation techniques. There has been an attempt in the use of web-based reporting in South Africa but most of these sites are characterised by heavy text which makes them time consuming to use or maintain. As a result most sites have not been updated or have ceased to exist because of the work load involved. The success of web reporting mechanisms in developed countries offers evidence that web-based reporting systems when appropriately visualised can improve the easy understanding of information and efficiency in the analysis of that data. In this thesis, a web-based reporting prototype was proposed after gathering information from different sources: literature related to disease reporting and the visualisation of infectious diseases; the exploration of the currently deployed web systems; and the investigation of user requirements from relevant parties. The proposed prototype system was then developed using Adobe Flash tools, Java and MySQL languages. A focus group then reviewed the developed system to ascertain that the relevant requirements had been incorporated and to obtain additional ideas about the system. This led to the proposal of a new prototype system that can be used by the authorities concerned as a plan to develop a fully functional disease reporting system for South Africa.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2015-12-07T18:39:22Z No. of bitstreams: 1 izabelamauricioderezende.pdf: 1559679 bytes, checksum: 67e4268b6adc9ef4deff3f2809a09f64 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2015-12-07T21:45:44Z (GMT) No. of bitstreams: 1 izabelamauricioderezende.pdf: 1559679 bytes, checksum: 67e4268b6adc9ef4deff3f2809a09f64 (MD5) / Made available in DSpace on 2015-12-07T21:45:44Z (GMT). No. of bitstreams: 1 izabelamauricioderezende.pdf: 1559679 bytes, checksum: 67e4268b6adc9ef4deff3f2809a09f64 (MD5) Previous issue date: 2015-02-10 / O Brasil é um país tropical de grande extensão territorial, com mais de um terço recoberto por florestas tropicais e outros ecossistemas naturais com condições ideais para a ocorrência de diversas arboviroses, as quais são mantidas em uma grande variedade de ciclos zoonóticos. Dentro deste cenário, Minas Gerais abriga um dos biomas mais importantes do Brasil, a Mata Atlântica, considerado ‘hotspot’, ou seja, região com uma rica biodiversidade. Estes fatores somados às diferenças climáticas, a grande variação latitudinal e as variadas tipologias vegetacionais do estado, propiciam a ocorrência de áreas com elevados índices de diversidade e endemismo de mamíferos. Diante deste contexto, este trabalho visou realizar a prospecção de flavivírus em culicídeos e pequenos mamíferos, em Minas Gerais, Brasil. Pools de culicídeos coletados em Montes Claros, no ano de 2012, tiveram seu RNA total extraído e foram testados por RT-PCR para a presença de flavivírus. Uma coleção de pequenos mamíferos, que foram coletados em Rio Pomba, MG, nos anos de 2012 e 2013, pertencentes a Coleção-ECOVIR, também foram analisados neste trabalho, sendo utilizados o soro de 115 animais e fígado de 54 animais desta coleção. Estas amostras passaram pelo processo de extração de RNA total, seguida da síntese de cDNA e testes para a presença de flavivírus através da técnica de qPCR. Dos 96 pools de culicídeos testados, um pool de A. aegypti pode estar naturalmente infectado com DENV-1. Posteriormente, 69 pools foram testados para YFV, SLEV e ROCV e um pool de A. scapularis apresentou fragmento de DNA com tamanho esperado para ROCV no PCR e apresentou um amplicon na reação de qPCR com iniciadores Flavi-1, indicando a circulação de flavivírus em Aedes scapularis no ambiente urbano de Montes Claros. Uma amostra de roedor Calomys sp. foi detectada como naturalmente infectada com flavivirus (possivelmente SLEV, DENV-1, DENV-3 e DENV-4), mostrando que roedores podem fazer parte da cadeia de manutenção e/ou transmissão de flavivírus na natureza, no Brasil. Esta amostra de roedor foi coletada na área de pasto, que poderia ser considerada uma área de transição entre o ambiente silvestre e peridoméstico. / Brazil is a tropical country of vast territory, with more than one third covered with tropical forests and ecosystems creating ideal conditions for the existence of many arboviruses that are maintained in a variety of zoonotic cycles. Within this scenario, Minas Gerais presents three of the most important biomes of Brazil, the Atlantic Forest that is considered hotspot of biodiversity. These factors plus the climatic differences, the great latitudinal range and the diversity of vegetation favor the occurrence of areas with high diversity index and endemism of mammals. Given this context, this work aims to prospect flaviviruses in mosquitoes and small mammals, in Minas Gerais, Brazil. Pools of mosquitoes collected in Montes Claros, in 2012, were submitted to RNA extraction and tested by RT -PCR for the presence of flaviviruses. A collection of small mammals (Collection- ECOVIR) collected in Rio Pomba, MG, in 2012 and 2013 was also analyzed in this work. A total of 115 serum samples and 54 liver samples were submited to RNA extraction, followed by cDNA synthesis and they were tested for the presence of flavivirus by qPCR technique. A total of 96 mosquito pools was tested and one pool of Aedes aegypti presented an amplicon that could represent a DENV-1 infection. Subsequently, 69 pools were tested for YFV, SLEV and ROCV and a pool of A. scapularis presented a DNA amplicon fragment with expected size for ROCV and it was positive in qPCR using primers Flavi-1, indicating the circulation flavivirus in Aedes scapularis in the urban environment, in Montes Claros. A sample of a rodent Calomys sp. was detected as naturally infected with flaviviruses (possibly SLEV, DENV-1, DENV -3 and DENV-4), indicating that rodents may be part of the maintenance chain and/or flavivirus transmission in nature, Brazil. This rodent sample was collected in the pasture area that could be considered a transition area between the wild and peridomestic environments.

O projeto educativo \"Para Viver de Bem com os Bichos\" foi implantado nas Unidades Educacionais do Município de São Paulo em 2002, destinado à difusão e promoção do conceito da posse responsável de animais de estimação, em especial de cães e gatos e desde então, trabalhou com professores de 1.605 escolas da cidade. O presente estudo foi delineado para analisar a dinâmica do processo educativo, verificar o impacto da metodologia empregada e o papel do professor como multiplicador no repasse das informações que compõem o projeto oferecido no ano de 2008. No primeiro momento foram avaliados os professores (G1) e em seguida os professores multiplicadores (G2) nas respectivas unidades de ensino. As avaliações foram feitas com a utilização de questões abertas e para a análise das respostas obtidas, utilizou-se o teste de McNemar. O conhecimento prévio dos professores sobre a posse responsável de animais de estimação mostrou-se insatisfatório quanto aos principais cuidados com animais, zoonoses e sua prevenção, cuidados pré e pós-agressão, método de controle reprodutivo e sua justificativa. O curso proporcionou a aquisição de conhecimento sobre os temas abordados, mas não foi suficiente para garantir a ação dos professores capacitados como instrumentos de repasse de informação técnica do projeto em sua unidade de ensino. Devem ser criados mecanismos de acompanhamento do desempenho do professor multiplicador em suas unidades de trabalho. Recomenda-se que políticas públicas sejam estabelecidas para facilitar a atividade educativa e diminuir os problemas que inviabilizam a possibilidade da teoria ser aplicada na prática. / The educative project \"How to live well with the animals\" was introduced in the Educational Units of the São Paulo Municipal District, in 2002, destined to diffusion and promotion of the concept of responsible pets ownership, in special dogs and cats and since then, it has worked with teachers of 1.605 schools of the city. The current study was written to analyze the dynamics of the educational process, to verify the impact of the used methodology and the role of the teacher as a multiplier in the repass of the information that make the project offered in the year of 2008. At the first moment the teachers evaluated (G1) and in the sequence the multiplier teachers (G2) in the respective teaching units. The evaluations were made with the use of open questions and for the analysis of the answers obtained they used the test of McNemar. The previous knowledge of the teachers about responsible possession of pets showed itself not satisfactory about the main cares with animals, diseases and its preventions, cares before and after aggression, method of reproductive control and its justifications. The course has promoted the acquisition of knowledge about the themes treated, but it was not enough to guarantee the action of the qualified teachers as the instruments for repassing the technical information of the project in their teaching units. Mechanisms to follow-up the performance of the multiplex teachers in the working units must be created. It is recommended that the public policies be established to facilitate the educational activities in order to diminish the problems that make unviable the application of the theory in the practice.

Dermatophytoses represent a major health burden in animals and man. Zoophilic dermatophytes usually show a high specificity to their original animal host but a zoonotic transmission is increasingly recorded. In humans, these infections elicit highly inflammatory skin lesions requiring prolonged therapy even in the immunocompetent patient. The correct identification of the causative agent is often crucial to initiate a targeted and effective therapy. To that end, matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) represents a promising tool. The objective of this study was to evaluate the reliability of species identification of zoophilic dermatophytes using MALDI-TOF MS. The investigation of isolates from veterinary clinical samples suspicious of dermatophytoses suggests a good MALDI-TOF MS based identification of the most common zoophilic dermatophyte Microsporum canis. Trichophyton (T.) spp. usually achieved scores only around the cutoff value for secure species identification because of a small number of reference spectra. Moreover, these results need to be interpreted with caution due to the close taxonomic relationship of dermatophytes being reflected in very similar spectra. In our study, the analysis of 50 clinical samples of hedgehogs revealed no correct identification using the provided databases, nor for zoophilic neither for geophilic causative agents. After DNA sequencing, adaptation of sample processing and an individual extension of the inhouse database, acceptable identification scores were achieved (T. erinacei and Arthroderma spp., respectively). A score-oriented distance dendrogram revealed clustering of geophilic isolates of four different species of the genus Arthroderma and underlined the close relationship of the important zoophilic agents T. erinacei, T. verrucosum and T. benhamiae by forming a subclade within a larger cluster including different dermatophytes. Taken together, MALDI-TOF MS proofed suitable for the identification of zoophilic dermatophytes provided fresh cultures are used and the reference library was previously extended with spectra of laboratory-relevant species. Performing independent molecular methods, such as sequencing, is strongly recommended to substantiate the findings from morphologic and MALDI-TOF MS analyses, especially for uncommon causative agents.

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Escherichia coli is part of the normal gastrointestinal microbiota of many animals, especially cattle. While most strains are commensal, Shiga toxin-producing E. coli (STEC) can cause severe human illness. Pathogenicity of STEC is associated with genes such as those encoding Shiga toxins, enterohemolysin, and intimin. By targeting these genes, highly sensitive molecular-based techniques help detect potentially harmful STEC. Persistent carriers and environmental contamination may be responsible for maintenance of STEC in cattle farms. Prevalence may be further influenced by diet, distance to contaminated water-sources, wildlife contact, slurry application to pasture, and population density. Relevance in environmental contamination is expected proportional to the amount of STEC shed in feces, but there is no consensus as to which production stage/age is most important. Distribution and transmission of STEC O157 are widely studied, but risk factors for non-O157 STEC are not as well defined. Understanding what contributes for contamination of animals prior to concentration in high-density feedlots may reveal opportunities for upstream control of shedding and transmission. Our purpose was to: (a) determine prevalence of STEC in fecal samples from animals in a cow-calf pasture-based production system; (b) describe effects of age class (dam, calf), spatial distribution of cattle, and time-point of sampling on distribution of strains positive for virulence genes stx1, stx2, eaeA, and hlyA; (c) isolate and identify serotypes present in stx-positive samples; and (d) assess genetic similarity of isolates. Understanding factors that influence distribution of STEC strains may help support on-farm management strategies with potential to yield safer beef products. / Master of Science

Thesis (PhD)-- Stellenbosch University, 2013. / ENGLISH ABSTRACT: The emergence and re-emergence of viral human pathogens from wildlife sources in the recent past has led to increased studies and surveillance of wildlife for potentially zoonotic agents in order to gain a better understanding of the pathogens, their sources as well as events that may lead to viral emergence. Of the >1407 known human pathogens, 13% are classified as emerging or re-emerging, and 58% as zoonotic; 37% of the (re-)emerging and 19% of the zoonotic pathogens are RNA viruses, accounting for the majority of recently emerged infectious diseases with a zoonotic origin, such as HIV, Ebola, Hendra, Nipah, Influenza and SARS. This study focusses on potentially zoonotic viruses hosted by rodents (Muridae family), shrews (order previously known as Insectivora/Soricomorpha, now reclassified as Eulipotyphla) and bats (order Chiroptera). Rodents and bats represent the largest (~40%) and second largest (~25%) mammalian orders and both occur on every continent except Antarctica. Together, the three mammalian orders investigated represent the most relevant potential sources of new zoonoses. In this study I investigated the occurrence of astroviruses, arenaviruses, coronaviruses and hantaviruses in South African small mammal species belonging to the orders mentioned above. These viruses have either been implicated in recent emerging zoonotic events or are considered to have the potential to cause cross-species transmissions resulting in a zoonotic event. In the first part of the study specimens collected from various bat, rodent and shrew species were screened for viral sequences by broadly reactive PCRs; positive samples were characterised by sequencing and sequence analysis. A separate part of the study focussed on hantavirus disease in humans: a seroprevalance survey was conducted to determine the presence of hantavirus antibodies in the local population. Additionally, acutely ill patients with potential hantavirus disease were tested in an attempt to identify possible acute infections and define clinical hantavirus disease in South Africa. Screening of rodent and shrew specimens resulted in the identification of eight novel arenavirus sequences. Seven of the sequences are related to Merino Walk virus, a recently identified South African arenavirus, and the eighth sequence represents a novel lineage of Old World arenaviruses. Screening of bat specimens resulted in the identification of highly diverse novel astrovirus and coronavirus sequences in various South African bat species, including the identification of a viral sequence closely related to the recently emerged Middle East Respiratory Syndrome coronavirus. While the study did not identify hantavirus infections in any of the acutely ill patients, it found seroprevalences similar to those observed in Europe and West Africa. The results obtained highlight the importance of small mammals in the emergence of potential zoonoses and further reinforce the importance of viral surveillance of relevant wildlife species. Further in-depth studies of naturally infected reservoir host populations are required in order to gain a better understanding of virus-host dynamics and the events that lead to virus emergence. / German Research Foundation (DFG) (project number: KR1293/9-1/13-1) / The Polio Research Foundation and the NHLS Research / Harry Crossley Foundation, the Polio Research Foundation and Stellenbosch University for granting scholarships and bursaries for PhD.

Cette thèse a pour objectif l'étude épidémiologique de zoonoses chez le sanglier en Corse afin d'éclairer l'analyse et la gestion du risque zoonotique. La première partie décrit la démarche d'analyse du risque et les données nécessaires à son estimation, met en évidence que le sanglier, porteur potentiel de nombreux agents pathogènes, est un modèle biologique intéressant pour le suivi de maladies transmissibles à l'Homme par ingestion ou manipulation de carcasses, et décrit le contexte d'étude, la Corse, notamment sur le plan cynégétique et de l'élevage. La seconde partie présente le dispositif mis en place pour la collecte des données, un réseau d'épidémiosurveillance active à l'échelle de la région insulaire, puis l'étude épidémiologique de trois agents zoonotiques (respectivement maladies) : Trichinella britovi (trichinellose), Toxoplasma gondii (toxoplasmose) et Mycobacterium bovis (tuberculose bovine). A l'issue de nos travaux, le risque de trichinellose, avéré en 2004, demeure difficile à évaluer mais plausible, nécessitant le maintien des consignes de prévention (cuisson de la viande, test des carcasses commercialisées). Le risque de toxoplasmose est fort sur l'ensemble de l'île et davantage encore dans les zones à forte densité d'exploitations agricoles. Le risque de tuberculose apparaît majeur à considérer, le bacille étant présent à la fois chez les sangliers et chez des porcs ou bovins de quatre régions corses. Nos travaux concluent en l'importance d'une meilleure gestion des carcasses et viscères d'animaux sanvages et domestiques, et la nécessité d'une coordination pérenne du dispositif mis en place pour le suivi des zoonoses en Corse.