Submitted by (edna.saturno@ufrpe.br) on 2016-10-14T12:55:53Z No. of bitstreams: 1 Joubert Santana Pimentel.pdf: 331586 bytes, checksum: a5ad502fe9d1ea2473615213205a4a88 (MD5) / Made available in DSpace on 2016-10-14T12:55:53Z (GMT). No. of bitstreams: 1 Joubert Santana Pimentel.pdf: 331586 bytes, checksum: a5ad502fe9d1ea2473615213205a4a88 (MD5) Previous issue date: 2009-03-02 / Modern zoological gardens are institutions to wild fauna maintenance, aiming its conservation, the performance of scientific research and leisure, recreation and environmental education activities. The variety of wild species living in conditions different from the one found in its natural habitats represents a propitious environment for diseases spread, specially the zoonotic ones. Due to data shortage and wild mammals’ epidemiological relevance both for toxoplasmosis as for leptospirosis, this study aimed to determine the seroprevalence of toxoplasmosis and leptospirosis in neotropical wild mammals, from “Zoológico do Parque da Cidade Governador José Rollemberg Leite”, Aracaju, Sergipe, Brazil. Blood samples were collected from 32 wild mammals, adults and from both sexes: 14 tufted capuchins (Cebus apella), four golden-bellied capuchins (Cebus xanthosternus), three pumas (Puma concolor), one jaguar (Panthera onca), one fox (Pseudolopex vetulus), six crab-eating raccoons (Procyon cancrivorus), two South American coatis (Nasua nasua), and one tayra (Eira Barbara). Sera were tested by the modified agglutination test (MAT) using formalin-fixed whole tachyzoites and mercaptoetenhol (cut-off point 1:25) to Toxoplasma gondii antibodies, and by the microscopic seroagglutination test (cut-off point: 1:100) using 24 serovar of,pathogenic leptospiras and 2 serovar of saprophyte leptospiras. Antibodies to T. gondii were found in 17 of 32 wild mammals (53.1%), and antibodies to Leptospiras spp were found in 4 of 32 wild mammals (12.5%). In relation to gender, 9 of 15 (60%) males, and 8 of 17 (47.1%)females had antibodies to T. gondii, and 4 of 15 (26.7%) males had antibodies to Leptospira spp. Considering the origin of wild seropositive mammals to T. gondii, 8 of 17 (47%) was born in zoo, 7 of 17 (41.2%) were from other zoos, and 2 (11.8%) was from the wild, and 3 of4 (75%) seropositive to Leptospiras spp was born in zoo, and 1 of 4 (25%) was from the wild. This is the first serological survey of Leptospiras spp in neotropical primates and carnivores from Northeast zoo from Brazil, and the first found to antibodies to Leptospira spp (serovar Copenhageni) in the threaten specie of primate golden-bellied capuchins (Cebus xanthosternus). / Os zoológicos modernos são instituições destinadas à manutenção da fauna selvagem com objetivo de conservação, pesquisa cientifica, lazer, recreação e educação ambiental. A ampla variedade de espécies selvagens, vivendo em condições diferentes do seu habitat natural, representa um ambiente propício à disseminação de doenças, muitas delas zoonóticas. Devido à escassez de dados e à relevância dos mamíferos selvagens no contexto epidemiológico, tanto na toxoplasmose, quanto na leptospirose, objetivou–se determinar a soroprevalência de toxoplasmose e de leptospirose em mamíferos selvagens neotropicais do Zoológico do Parque da Cidade Governador José Rollemberg Leite, Aracaju, Sergipe. Para tanto foram colhidas amostras sanguíneas de 32 mamíferos selvagens, adultos e de ambos os sexos, das espécies: 14 macacos-prego (Cebus apella), quatro macacos-prego-do-peito-amarelo (Cebus xanthosternus), três onças-suçuaranas (Puma concolor), uma onça-pintada (Panthera onca), uma raposa-do-campo (Pseudolopex vetulus), seis guaxinins (Procyon cancrivorus), dois quatis (Nasua nasua) e um papa-mel (Eira barbara). Para pesquisa de anticorpos anti- Toxoplasma gondii foi utilizado o teste de Aglutinação Modificada com taquizoítos inativados na formalina e 2-mercaptoetanol (ponto de corte na diluição 1:25) e para pesquisa de anticorpos anti-Leptospira spp foi utilizado o teste de Soroaglutinação Microscópica (ponto de corte na diluição 1:100) com uma coleção de antígenos vivos que incluiu 24 variantes sorológicas de leptospiras patogênicas e duas leptospiras saprófitas. Dentre os 32 mamíferos, 17 (53,1%) apresentaram anticorpos anti-T. gondii e quatro (12,5%) foram positivos para anticorpos anti-Leptospira spp. De acordo com o sexo, 60% (9/15) dos machos e 47,1% (8/17) das fêmeas foram soropositivos para T. gondii e 26,7% (4/15) dos machosapresentaram anticorpos anti-Leptospira spp. Com relação à procedência, dos mamíferos que apresentaram anticorpos anti-T. gondii, 47% (8/17) nasceram no zoológico, 41,2% (7/17) foram oriundos de outras instituições e dois (11,8%) foram provenientes da natureza. Em relação aos quatro mamíferos soropositivos para Leptospira spp, três (75%) foram procedentes da natureza e um (25%) nasceu no zoológico. Este trabalho representou o primeiro inquérito sorológico para a ocorrência de anticorpos anti-Leptospira spp em primatas e carnívoros neotropicais em um zoológico do Nordeste do Brasil e descreveu pela primeira vez a ocorrência de anticorpos anti-T. gondii e anti-Leptospira spp com sorovar mais provável Copenhageni no primata ameaçado de extinção macaco-prego-de-peito-amarelo (C. xanthosternus).

Submitted by (edna.saturno@ufrpe.br) on 2016-10-20T16:38:14Z No. of bitstreams: 1 Natalia Matos Souza.pdf: 205714 bytes, checksum: 59b167a505e1216282eec9736dde0f7d (MD5) / Made available in DSpace on 2016-10-20T16:38:14Z (GMT). No. of bitstreams: 1 Natalia Matos Souza.pdf: 205714 bytes, checksum: 59b167a505e1216282eec9736dde0f7d (MD5) Previous issue date: 2012-02-20 / This study was aimed to research Leptospira spp agglutinins in horses in the region of the forest area and Fernando de Noronha island in the state of Pernambuco; identifying highest areas of leptospirosis incidence; identifying the serovars that occur more frequently in the studied areas; relating clinical signs with the agglutinins presence in blood serum of equines; observing the reproductive disorders connection with the agglutinins presence in the equines blood serum; relating the presence of rodents and seropositive animals; observing the occurrence of anti leptospiras antibodies at different age groups. It was collected blood from 328 equines of both genders, aging beyound 6 months. After material centrifugation the serum was analyzed by LANAGRO/PE through the microscopic agglutination test. Of the 328 collected samples were found 84 reagents animals in the microscopic agglutination test for leptospirosis. Among which 12 (1.5%) foals, 20 (25.64%) young, 42 (31.11%) adults and 10 (28.57%) senile. Of the 84 reagents animals 69.04% (58) are female and 30.95% (26) are male; 3.75% (3) are Campolina, 2,3% (2) are mestizos, 37,71% (30) are of the breed Quarter horse and 58,33% (49) are Mangalarga Marchador. Among the studied counties Aldeia stood out to present the highest percentage of reagent animals with 32,31% (65) subsequently Pombos with 28,2% (22), Palmares with 23,81% (5), Bonanza with 22,36% (17), Paudalho with 22% (11) and finally Escada with 14,3%(4). The only studied county that did not present any reagent animal was Itambé. Among the 27 researched serovars by the microscopic seroagglutination test for Leptospirosis diagnostic, 10 were found in reagent animals, which, in order of occurrence Icterohaemorrhagiae (58), Shermani (26), Panama (15), Copenhageni (10), Batavie (3), Hardjo (2), Australis (1), Pyrogenes (1), Cynopteri (1) e Canicola (1). In this study 3 of 16 analyzed samples, 10 were considered reagent and 6 were considered non-reagent. 62,5% of samples were positives to the serovar Icterohaemorrhagiae, 37,7% were considered positive to serovar Copenhageni. Realizing this study it’s possible to conclude that leptospirosis in equines occurs in endemic way at the studied area, considering that of 328 analyzed samples 84 (25,62%) were positives to the test. The presence of rodents was very important to the appearance of condition as proved how it was related in all visited property. From this study it was possiblited trace an epidemiologic profile of the leptospirosis in equines belonging to Zona da Mata and Fernando de Noronha island of the state of Pernambuco. Knowing the risk factors involving this condition and which may be useful to treat and prevent, however it’s still needed a extensive study to determinate the reason of the occurrence of certain serovars in some studied regions. / Objetivou-se com esse trabalho realizar uma pesquisa de aglutininas anti Leptospira spp em equinos na região da zona da mata e ilha de Fernando de Noronha no Estado de Pernambuco; identificar áreas de maior ocorrência da leptospirose; identificar os sorovares que ocorrem com maior frequência nas áreas estudadas; relacionar os sinais clínicos com a presença de aglutininas no soro sanguíneo dos equinos; relacionar a presença de roedores com a presença de animais soropositivos; observar a ocorrência de anticorpos anti leptospiras nas diferentes faixas etárias. Coletou-se sangue de 328 equinos de ambos os sexos, com idade a partir dos seis meses. Após centrifugação do sangue o soro foi analisado pelo teste de soro aglutinação microscópica, com ponto de corte na diluição de 1:100. Das 328 amostras examinadas constatou-se que 84 (25,6%) animais foram reagentes para aglutinação anti-Leptospira spp. Dentre os quais 12 (1,5%) potros, 20 (25,64%) jovens, 42 (31,11%) adultos e 10 (28,57%) senis. Dos 84 animais reagentes 69,04% (58) são fêmeas e 30,95% (26) são machos; 3,57% (3) são da raça Campolina, 2,3% (2) são mestiços, 35,71% (30) são da raça Quarto de Milha e 58,33% (49) são da Raça Mangalarga Marchador. Dentre as regiões estudadas Aldeia se destacou por apresentar o maior percentual de animais reagentes, com 32,31% (65), em seguida Pombos com 28,2% (22), Palmares com 23,81% (5), Bonanza 22,36% (17), Paudalho 22% (11) e por fim, Escada com 14,3% (4). Dentre os 27 sorovares pesquisados 10 foram mais prevalentes, em ordem de maior ocorrência, Icterohaemorrhagiae (58), Shermani (26), Panama (15), Copenhageni (10), Batavie (3), Hardjo (2), Australis (1), Pyrogenes (1), Cynopteri (1) e Canicola (1). Na ilha de Fernando de Noronha das 16 amostras analisadas 10 foram consideradas reagentes e 6 foram consideradas não reagentes. Considerando os sorovares mais prevalentes 62,5% das amostras foram positivas para o sorovar Icterohaemorrhagiae, 37,5% foram positivas para o sorovar Copenhageni. Com a realização desse estudo foi possível concluir que a leptospirose na espécie equina ocorre de forma endêmica nas áreas estudadas. Acredita-se que a presença de roedores foi importante no aparecimento da afecção haja vista que foi relatada sua presença em todas as propriedades visitadas, todavia necessita-se de maiores estudos para verificar a participação destes animais na transmissão da leptospirose aos equinos. A partir desse trabalho pode-se traçar um perfil soro epidemiológico da leptospirose em equinos na Zona da Mata e na ilha de Fernando de Noronha no Estado de Pernambuco, que pode ser útil para realização de tratamento e da prevenção desta enfermidade, todavia, ainda é preciso estudos mais aprofundados para determinarmos o porque da ocorrência de determinados sorovares em algumas regiões estudadas.

Made available in DSpace on 2014-08-20T13:32:56Z (GMT). No. of bitstreams: 1 tese_daiane_drawanz_hartwig.pdf: 4007239 bytes, checksum: 7d50e5824c8c66e23049bd005ad56ea6 (MD5) Previous issue date: 2010-12-20 / Leptospirosis is a serious infectious disease caused by pathogenic spirochetes of the genus Leptospira, it is classified as a zoonosis of worldwide distribution. This disease results morbidity and mortality in humans and animals, justifying the application of prophylactic strategies. Current vaccines against leptospirosis are composed of inactivated bacteria and do not stimulate cross-protection. Thus, there is need to develop a safe and effective vaccine. In this study, we used the outer membrane proteins LigANI e LipL32, because they have been identified as vaccinogens. These, in their recombinant form, are usually expressed in Escherichia coli and as subunit vaccines have shown variable efficacy. We describe in this work the use of Pichia pastoris as an alternative expression system. The genes ligANI and lipL32 were cloned into vector pPICZαB, which allowed the secretory expression of proteins in P. pastoris. The protein yield in this system was 276 mg/L for LigANI and 285 mg/L for LipL32. The recombinant proteins were glycosylated and remained antigenic. The immunoprotective potential was evaluated in the hamster model, challenged with virulent L. interrogans serovar Copenhageni. Both proteins induced high levels of antibodies (P < 0.001). The animals immunized with LigANI and LipL32 using aluminium hydroxide as adjuvant, showed no protection against challenge, but showed a significant increase in survival (P < 0.001). In conclusion, the yeast P. pastoris has proved an efficient heterologous expression system of LigANI and LipL32 L. interrogans proteins. The secreted and glycosylated LigANI protein may be used in the control of leptospirosis, although additional studies are needed. / Leptospirose é uma doença infecciosa grave causada por espiroquetas patogênicas do gênero Leptospira, sendo classificada como uma zoonose de ampla distribuição mundial. Esta doença resulta morbidade e mortalidade em humanos e animais, justificando a aplicação de estratégias profiláticas. As vacinas atuais contra a leptospirose são compostas por bactérias inativadas e não estimulam proteção cruzada. Assim, existe a necessidade de desenvolver uma vacina efetiva. No presente estudo, as proteínas de membrana externa LigANI e LipL32 foram utilizadas, pois são apontadas como potenciais vacinógenos. Estas, em sua forma recombinante, costumam ser expressas em Escherichia coli e, como vacina de subunidade tem apresentado eficiência variável. Nós descrevemos neste trabalho a utilização da levedura Pichia pastoris como sistema de expressão alternativo. Os genes ligANI e lipL32 foram clonados no vetor pPICZαB, que permitiu a expressão secretória das proteínas em P. pastoris. O rendimento das proteínas neste sistema foi de 276 mg/L para LigANI e 285 mg/L para LipL32. As proteínas recombinantes foram glicosiladas e mantiveram-se antigênicas. O potencial imunoprotetor das proteínas foi avaliado em modelo hamster desafiado com cepa virulenta de L. interrogans sorovar Copenhageni. Ambas as proteínas induziram altas taxas de anticorpos (P < 0,001). Os animais imunizados com LigANI e LipL32, utilizando hidróxido de alumínio como adjuvante, não apresentaram proteção contra o desafio, mas demonstraram um aumento significativo na sobrevida (P < 0,001). Em conclusão, a levedura P. pastoris demonstrou ser um eficiente sistema de expressão heterólogo das proteínas LigANI e LipL32 de L. interrogans. A proteína LigANI secretada e glicosilada pode ser utilizada no controle da leptospirose, embora estudos adicionais sejam necessários.

Made available in DSpace on 2014-08-20T13:32:57Z (GMT). No. of bitstreams: 1 dissertacao_andre_grassmann.pdf: 1497268 bytes, checksum: 73f55a329ef2c9be70227b63b45785a2 (MD5) Previous issue date: 2011-02-28 / Leptospirosis is an infectious disease that affects humans, wild and domestic animals worldwide. Pathogenic spirochetes from the Leptospira genus are the causative agents of this zoonosis. The several Leptospira species have noted antigenic diversity, even within the same species. This is the main reason current bacterin vaccines have limitations, such as adverse effects and short term immunity, restricting their use in human populations. The need for effective leptospirosis vaccines promoted studies on characterization of new vaccine candidates. The 32 kDa outer membrane lipoprotein, LipL32, is the most abundant protein in the whole leptospira proteome, it is conserved in all pathogenic serovars and absent in saprophytes. This protein is immunogenic and able to bind to mammalian extracellular matrix. However, LipL32 subunit vaccines did not protect animals against challenge. In an attempt to solve this, we use LipL32 fused and coadministered with B subunit of the Escherichia coli heat-labile enterotoxin (LTB) to enhance the immune response. LTB is a non-toxic molecule with immunoestimulatory and immunomodulatory properties. The recombinant proteins rLTB, rLipL32 and rLTB::LipL32 were expressed in E. coli, purified and characterized. Female hamsters were distributed in groups as follows: rLTB; rLTB+rLipL32; rLTB::LipL32, homologous bacterin; PBS. The serum from each animal was collected for humoral immune response determination by ELISA. The animals were challenged with 5×LD50 dose of Leptospira interrogans strain Fiocruz L1-130. Both treatments induced high titers of anti-rLipL32 antibodies. The rLTB+rLipL32 and rLTB::LipL32 treatments afforded significant protective response upon challenge, when compared to control groups (p<0.05). No prior study with leptospirosis had used LTB as the adjuvant, or fused antigens in an attempt to control this disease. Furthermore, this is the first report of a protective subunit vaccine using rLipL32 as the antigen, and an important contribution towards the development of improved leptospirosis vaccines. / A leptospirose é uma doença infecciosa que afeta humanos e animais silvestres e domésticos em todo mundo. As espiroquetas patogênicas do gênero Leptospira são os agentes causadores desta zoonose. As diversas espécies de leptospiras possuem notada diversidade antigênica, inclusive em uma mesma espécie. Esta característica resulta em limitação das atuais vacinas bacterinas que não induzem proteção cruzada entre os diferentes sorovares. Além disso, estas vacinas geram efeitos adversos e imunidade de curta duração, restringindo seu uso em populações humanas. A necessidade de novas vacinas eficazes contra a leptospirose estimulou estudos para caracterizar novos antígenos vacinais. A lipoproteína de membrana externa de 32 kDa, LipL32 é a proteína mais abundante no proteoma total da leptospira, conservada entre todos os sorovares patogênicos e ausente nas leptospiras saprófitas. Esta proteína é imunogênica e possui habilidade de ligar-se à matriz extracelular de mamíferos. Porém, animais inoculados com vacinas de subunidade utilizando LipL32 não sobrevivem ao desafio. Em função disso, utilizamos LipL32 fusionada e co-administrada com a subunidade B da enterotoxina termolábil de Escherichia coli (LTB) para melhorar a resposta imune. LTB é uma molécula atóxica com reconhecida atividade imunoestimuladora e imunomoduladora. As proteínas recombinantes rLTB, rLipL32 e rLTB::LipL32 foram produzidas em E. coli, purificadas e caracterizadas. Hamsters fêmeas foram distribuídas em grupos e inoculadas com duas doses, da seguinte forma: rLTB; rLTB+rLipL32; rLTB::LipL32, bacterina homóloga e PBS. Soro foi coletado individualmente para determinação da resposta imune humoral por ELISA. Os animais foram desafiados com uma dose de 5×DL50 de Leptospira interrogans cepa Fiocruz L1-130. Os tratamentos induziram altos títulos de anticorpos anti-rLipL32. Os tratamentos rLTB+rLipL32 e rLTB::LipL32 induziram resposta protetora significativa frente ao desafio quando comparados com os grupos controle (p<0,05). Nenhum estudo anterior usou LTB como adjuvante para uma vacina contra leptospirose, tampouco antígenos fusionados com o intuito de controlar esta doença. Além disso, este é o primeiro relato de indução de imunidade protetora utilizando rLipL32 como vacina de subunidade, uma importante contribuição para o desenvolvimento de vacinas mais eficazes contra leptospirose.

Made available in DSpace on 2014-08-20T13:33:00Z (GMT). No. of bitstreams: 1 tese_leonardo_garcia_monte.pdf: 839733 bytes, checksum: 46a2c729d1764535d017cab21dde7a45 (MD5) Previous issue date: 2011-12-16 / The development of reagents for infectious disease diagnosis and prevention is strategic for the scientific and technological advance of the country. Promising results have been obtained with the use of magnetic nanocomposites (MNC) in health for several purposes as separation, purification and detection of cells or biomolecules. In this work, an immunomagnetic separation (IMS) method using carbon and cobalt-based (MNCc), synthesized, and other polystyrene and iron-based (MNCp) commercially available was applied, for the isolation and detection of pathogenic leptospires. The MNCc were synthesized through polymeric precursor method and characterized by scanning electron microscopy/transmission (SEM/TEM), X-ray diffractometry (XRD), infrared spectrometry (FTIR), and immunofluorescence (IF). The MNCc surface was functionalized with carboxylic groups through the incorporation of acrylic acid. MNCc, adsorbed with monoclonal antibodies (MAbs) against pathogenic leptospires, were used in the IMS assays followed by PCR and cell culture was able to detect and isolate the organism. MNCp were used for detecting Leptospira spp. in biological fluids from dogs artificially contaminated with leptospires, and clinical samples from dogs positive for leptospirosis. The introduction of IMS, as a step prior to PCR using MNCp, detected a concentration of leptospires ten times smaller when compared to PCR performed directly on artificially contaminated urine and blood samples. Moreover, the IMS performed with MNCp increased PCR sensitivity in clinical samples from dogs positive for leptospirosis. Data from both studies suggest that MNCc and MNCp are important tools for the development of new diagnosis tests. / O desenvolvimento de insumos para o diagnóstico e prevenção de doenças infecciosas é estratégico para o avanço científico e tecnológico do país. Resultados promissores vêm sendo obtidos com o uso de nanocompósitos magnéticos (NCM) aplicados à área da saúde para as mais diversas finalidades, como na separação, purificação e detecção de células ou biomoléculas. Nesse trabalho, um método de imunoseparação magnética (IMS) utilizando NCM a base de carbono e cobalto (NCMc) sintetizados, e outro de poliestireno e ferro (NCMp) disponível comercialmente, foi utilizado para capturar e detectar leptospiras patogênicas. Os NCMc foram sintetizados através do método dos precursores poliméricos e caracterizados por microscopia eletrônica de varredura/transmissão (MEV/MET), difratometria de raio-X (DRX), espectrometria no infravermelho (FTIR) e imunofluorescência (IF). A superfície dos NCMc foi funcionalizada com grupos carboxílicos mediante incorporação de ácido acrílico. Os NCMc adsorvidos com anticorpos monoclonais (MAbs) contra leptospiras patogênicas, quando utilizados em ensaios de IMS seguidos por PCR e cultivo celular, foram capazes de detectar e isolar o organismo. Os NCMp foram utilizados para a detecção de Leptospira spp. em fluidos biológicos de cães artificialmente contaminados com leptospiras e em amostras clínicas de cães positivos para a leptospirose. A introdução da IMS como passo anterior a PCR utilizando os NCMp foi capaz de detectar uma concentração 10 vezes menor de leptospiras quando comparada a PCR realizada diretamente a partir de amostras de urina e sangue artificialmente contaminados. Além disso, a IMS realizada com os NCMp aumentou a sensibilidade da PCR em amostras clínicas de cães positivos para leptospirose. Os dados obtidos em ambos os estudos sugerem que os NCMc e NCMp constituem importantes ferramentas para o desenvolvimento de novos testes de diagnóstico.

Leptospirosis is a disease of global importance with a changing epidemiology in both humans and animals. It is also a significant zoonosis particularly in the developing world. To date there is limited knowledge of the incidence of leptospirosis in dogs in South Africa. This study was undertaken on a subset of dogs in South Africa to determine the presence of leptospiral antibodies to serovars known to infect dogs. Serum samples from both stray and owned dogs from various parts of South Africa were collected and tested against fifteen serovars of Leptospira. Five hundred and thirty samples were tested and twenty-five tested positive to seven different serovars. Nine of the 25 samples tested positive to more than one serovar. The two serovars that were most frequently represented were L. Canicola, which reacted to seventeen sera, and L. Pyrogenes, which reacted to nine sera in all. Currently the only vaccines available in South Africa in different combinations contain either L. Canicola, L. Icterohaemorrhagiae, L. Pomona or L. Grippotyphosa. The results show that the use of vaccines containing L. Canicola is still justifiable in certain regions of the country. However, the presence of antibodies to L. Pyrogenes in several dogs indicates that there is a need to investigate for the presence of antibodies in a larger group of dogs. This would allow vaccine manufacturers to tailor the Leptospira antigens present in vaccines to include those that are prevalent in a particular region or country. Copyright / Dissertation (MSc)--University of Pretoria, 2009. / Veterinary Tropical Diseases / unrestricted

Submitted by Cássio da Cruz Nogueira (cassionogueirakk@gmail.com) on 2017-05-15T18:03:30Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_InvestigacaoSoroepidemiologicaBrucelose.PDF: 308279 bytes, checksum: 3a3d80463e7c149e1dcc7d535ddca8bd (MD5) / Approved for entry into archive by Edisangela Bastos (edisangela@ufpa.br) on 2017-05-22T15:56:04Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_InvestigacaoSoroepidemiologicaBrucelose.PDF: 308279 bytes, checksum: 3a3d80463e7c149e1dcc7d535ddca8bd (MD5) / Made available in DSpace on 2017-05-22T15:56:04Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_InvestigacaoSoroepidemiologicaBrucelose.PDF: 308279 bytes, checksum: 3a3d80463e7c149e1dcc7d535ddca8bd (MD5) Previous issue date: 2007-06-18 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O objetivo do trabalho foi a detecção de anticorpos anti - Brucella abortus e anti – Leptospira interrogans em soros de eqüídeos e seus tratadores nos bairros das cidades de Belém e Ananindeua, abrangendo os meses de abril a agosto de 2005, utilizando para este fim, 195 soros sanguíneos de eqüídeos e 70 soros sanguíneos de homens que manipulavam os animais direta ou indiretamente. Para a pesquisa de animais sororeagentes à B. abortus, foram usadas as provas do Antígeno Acidificado Tamponado (AAT) como teste de triagem e a Soro Aglutinação Lenta em Tubos (SAL) e o teste do 2-Mercaptoetanol (2-ME), como teste confirmatórios. Para a Leptospirose, foi utilizada a prova de Soroaglutinação Microscópica (SAM), sendo realizada a triagem dos soros frente à 25 sorovares de L. interrogans, considerando-se positivos aquelas amostras com titulação igual ou maior que 100. De 195 amostras de soros sanguíneos de eqüídeos, 184 (94,87%) foram positivas para todos os sorovares analisados, sendo que os mais frequentemente encontrados foram: Patoc, Automnalis, Ictehaemohragiae, Pyrogenes e Bratislava. Para as amostras sanguíneas de homens, a positividade foi de 49 (70%) de soros reagentes, com os sorovares Patoc, Ictehaemohragiae, Bratislava, Butembo, Copenhageni e Automnalis os mais detectados. Das amostras positivas de animais e seus respectivos tratadores, 47/70 (67,14%) foram semelhantes para os mesmos sorovares de Leptospira spp., sendo que 2/70 (2,86%) amostras foram negativas em ambos os grupos pesquisados, 2/70 (2,86%) foram somente positivas em homens e 19/70 (27,14%) foram exclusivamente positivas nas amostras de soros de eqüídeos. Os bairros do Coqueiro, Guamá, 40 horas, Barreiro e Bengui apresentaram a maior percentagem de casos soropositivos. Não houve diferença significativa em relação às outras variantes estudadas, como: idade (animal e homem), tempo de serviço (animal e homem), espécie do animal, escore corporal do animal e grau de instrução do homem. Tanto nos animais quanto nos homens não foram detectadas reações positivas para B. abortus. / The objective of the work was the detention of antibodies anti - Brucella abortus and anti - Leptospira interrogans in serum of equines and equine workers of the quarters of the cities of Belém and Ananindeua, enclosing the months from April to August of 2005, using for this end, 195 sanguineous serum of eqquines and 70 sanguineous serum of men that manipulated the animals direct or indirectly. For the research of reagents serum of animals to B. abortus, the tests used had been Antigen Acidified Test (AAT), as a selection test, and slow seroagglutination (SAL) and and the 2-mercaptoetanol (2-ME) as confirmatory tests. To Leptospirosis, the test used was of microscopical seroagglutination being carried through the selection of the serum front to the 25 serovars of L. interrogans, considering positive those samples with titulation equal or bigger that 100. Of 195 samples of equine sanguineous serum, 184 (94.87%) had been positive for all serovars analyzed, being that more frequent found: Patoc, Automnalis, Ictehaemohragiae, Pyrogenes and Bratislava. For the sanguineous samples of men, the positivity was of 49 (70%) of reacting serum, with the most detected serovars: Patoc, Ictehaemohragiae, Bratislava, Butembo, Copenhageni and Automnalis. Of the positive samples of equine and its respective workers, 47/70 (67.14%) were similar for same serovars of Leptospira spp., being that 2/70 (2.86%) samples had been negative in both the searched groups, 2/70 (2.86%) were only positive in humans and 19/70 (27.14%) were exclusively positive in the samples of equine serum. The quarters of Coqueiro, Guamá, 40 horas, Barreiro and Bengui had presented the biggest percentage of positive cases serum. It did not have significant difference in relation to the other studied variants, as: climate, age (animal and man), time of service (animal and man), species of the animal, props up corporal of the animal and degree of instruction of the man. As much in the animals how much in the men positive reactions for B. abortus had not been detected.

Made available in DSpace on 2014-08-20T14:37:54Z (GMT). No. of bitstreams: 1 tese_clairton_marcolongo_pereira_resumo.pdf: 10803 bytes, checksum: 3f7507f184a7adb5ceb1296595a35eca (MD5) Previous issue date: 2014-02-21 / This thesis is a study developed in period of 34 years about equine diseases diagnosed through out 34 municipalities of Southern Rio Grande do Sul, which includes de influence area of the Laboratório Regional de Diagnóstico of the Veterinary School of the Federal University of Pelotas. There is a general paper about the diseases diagnosed in this specie related by etiologic agent. This paper establishes the importance of dermatological diseases, including equine sarcoid and squamous cell carcinoma as a cause of economic losses in equines. It also demonstrates that leucoencephalomalacia, rabies, thromboembolism by Strongylus vulgaris and equine monocytic ehrlichiosis are the leading causes of death in horses in the region. Other three papers are presented: two related to the causes of abortion and the one about the epidemiology of pythiosis. These conditions are important causes of economic losses in the breeding of horses in the region. / Esta tese trata-se de um estudo das enfermidades de equinos diagnosticadas em 34 municípios da região Sul do Rio Grande do Sul, que compreende a área de influência do Laboratório Regional de Diagnóstico da Faculdade de Veterinária da Universidade Federal de Pelotas, em um período de 34 anos. É apresentado um trabalho geral das enfermidades diagnosticadas nesta espécie animal, relacionadas por agente etiológico tendo sido estabelecida a importância das doenças dermatológicas, dentre elas o sarcoide equino e o carcinoma de células escamosas como causas de prejuízos econômicos nesta espécie animal. Com este estudo foi possível, ainda, demonstrar que a leucoencefalomalacia, a raiva, o tromboembolismo por Strongylus vulgaris e a erliquiose monocítica são as principais causas de morte de equinos na região. São apresentados, também, dois trabalhos científicos referentes às causas de aborto e um à epidemiologia da pitiose, respectivamente, que são importantes causas de prejuízos econômicos na criação de equinos na região, identificadas neste estudo.

La leptospirose est une zoonose causée par les bactéries du genre Leptospira. Elle touche près de 1 million d'individus par an dans le monde entier et sévit à l'état endémique dans les pays au climat tropical tel que La Réunion. Les manifestations habituelles sont variables d'un individu à l'autre et englobent une simple fièvre jusqu'aux défaillances poly-viscérales avec mortalité dans 5 à 10% des cas. Sa physiopathologie est encore mal comprise, en particulier la part que joue une réponse immunitaire inappropriée dans la genèse des manifestations graves qui surviennent en quelques heures, et avant la mise en place d'une réponse immunitaire adaptative propre à éliminer le microorganisme. Si l'échappement de la bactérie au système du complément est bien documenté, le rôle des acteurs cellulaires du système immunitaire inné reste à étayer. Notre étude avait donc pour objectif d'explorer l'immunopathologie de la leptospirose humaine dans la phase initiale de l’infection. Notre méthodologie s'est appuyée principalement sur des analyses quantitatives et qualitatives des acteurs cellulaires du système immunitaire inné à partir de prélèvements sanguins en phase précoce de la maladie, et comparaison avec la phase de convalescence et des sujets contrôles. Dans un premier temps nous avons montré qu'une population particulière de lymphocytes T impliquée dans la réponse immune innée, les lymphocytes Tγδ, avaient un taux abaissé et que cette baisse était corrélée à la charge bactérienne ainsi qu'à l’intensité de l'atteinte hépatique classiquement retrouvée lors de la leptospirose. Dans un deuxième temps, nous avons analysé les polynucléaires neutrophiles circulants dont le taux augmente d’autant plus que la maladie est sévère, mais sans pour autant présenter de modification de certains marqueurs d’activation ou de recrutement tissulaire (CD15, CD11b, CD182). Une des principales chimiokines des neutrophiles, l'interleukine 8, était à taux peu élevés. Les derniers travaux concernent les principales formes solubles issues des molécules membranaires impliquées dans le processus de recrutement/diapédèse leucocytaire. Nous retrouvons de manière isolée une forte élévation des formes solubles d'E-sélectine et ICAM-1 qui sont notamment exprimées par les cellules endothéliales. Ces augmentations n'étaient pas corrélées aux marqueurs de gravité de la maladie. La signification biologique de cette élévation n’est pas encore connue lors de la leptospirose. L'ensemble de nos données permet d’apporter des informations nouvelles sur des acteurs du système immunitaire inné présents dans le compartiment vasculaire lors de la leptospirose humaine. Cette réponse immunitaire semble inadaptée pour permettre une clairance du pathogène au stade de dissémination hématogène. / Leptospirosis is a bacterial zoonosis caused by Leptospira and affecting 1 million people each year worldwide and mainly in tropical areas such as Reunion Island. Usual presentations encompass flu-like syndrome to multiorgan failure with mortality rate between 5 to 10%. To date, pathophysiology in humans is poorly understood, notably the capacity of innateimmunity to mount a robust response to clear pathogen or to induce tissue damages and contributing to disease severity. Our study aimed at assessing the role of innate immune cells and molecules within the first days of leptospiral infection.Using blood samples, we performed quantitative and qualitative assessment of circulating innate immune cells from leptospirosis cases and healthy controls. The first study explored the levels of gamma-delta T-cells (γδT-cells), a subset of unconventional T cells with innate immune functions. Gamma-delta T cells were found deeply decreased and levels wereinversely correlated to bacterial burden and liver damage. The second study focused on membrane bound receptors indicative of activation and tissue migration ability of neutrophil polymorphonuclear cells: CD15, CD11b, and CD182. Although neutrophil rates were high in leptospirosis cases, the levels of studied receptors were either lower (CD15) or identical to healthy controls (CD11b, CD182). In addition, only low levels of interleukin-8, a key chemokine for neutrophils, was detected in patients. Lastly, we ascertained the plasmatic levels of several shed cell adhesion molecules notably expressed by endothelial cells. The levels of soluble E-selectin and ICAM-1 were significantly increased compared to controls, while P-selectin level was lower. We did not find any correlation with disease severity or organ failure. This finding indicates that endothelial cell may be activated but further experiments are warranted to explain the functional impact of our findings. Altogether, our results add to the field of knowledge of leptospirosis pathophysiology, and in particular the implication of key innate immune cells at the stage of plasmatic bacterial dissemination. Our findings will support the view that there is an inappropriate immune response to Leptospira.

La leptospirose est considérée comme la zoonose la plus répandue au monde mais les incidences sont les plus élevées dans les régions tropicales et en particulier sur les îles. Les îles du Sud-Ouest de l'Océan Indien ne dérogent pas à la règle puisque la maladie y représente un problème de santé humaine majeur sur plusieurs îles, notamment aux Seychelles qui enregistrent un des plus fort taux d'incidence humaine au monde. Sur la base des données disponibles, l'épidémiologie humaine apparait contrastée à l'échelle de la région : les cas cliniques sur Mayotte résultent d'infections par quatre espèces de leptospires distinctes alors qu'à La Réunion ou aux Seychelles, une seule espèce est à l'origine de la grande majorité des cas cliniques. L'objectif général de cette thèse est d'identifier certains des déterminants de cette épidémiologie singulière. Nous avons dans un premier temps complété les informations humaines en investiguant la leptospirose en Union des Comores, pays n'ayant jamais rapporté de transmission autochtone. Nos résultats indiquent que les populations humaines y sont exposées à des antigènes de leptospires comparables à ceux retrouvés sur l'île voisine de Mayotte. Ce résultat suggére que l'absence de leptospirose sur certaines îles est le résultat d'un déficit de surveillance. Nous avons ensuite caractérisé la diversité génétique des leptospires au sein de la faune de certaines îles, caractérisées par des niveaux d'endémisme élevés à même d'être en partie à l'origine de cette épidémiologie contrastée. Nous décrivons d'une part une importante diversité des leptospires pathogènes chez les chauves-souris (Chiroptères) malgaches. Nous montrons d'autre part que cette diversité de leptospires n'est pas structurée géographiquement mais présente au contraire une importante spécificité d'hôte, résultant de différents processus évolutifs incluant co-spéciation et host-switch. Nous avons exploité cette spécificité d'hôte pour éclairer l'épidémiologie de la leptospirose à Mayotte, où nous montrons que l'importante diversité bactérienne impliquée dans les cas cliniques résulte de la présence de nombreux réservoirs, dont certains originaires de Madagascar. Ainsi, il apparaît que l'épidémiologie humaine de la leptospirose dans le SOOI est le reflet d'assemblages distincts de leptospires cosmopolites et autochtones/endémiques maintenus et excrétés par des réservoirs animaux particuliers. / Leptospirosis is considered as the most widespread zoonosis worldwide but the incidence levels are higher in tropical regions and particularly on islands. The South-Western Indian Ocean islands are no exception and the disease is of major medical concern in several islands notably in Seychelles, displaying some of the highest human incidence ever reported. Based on available data, the human epidemiology appears contrasted in the region: on Mayotte, human cases result from the infection with four distinct Leptospira species whereas on Reunion Island or Seychelles, a single species causes the majority of clinical cases. The main objective of this thesis is to identify some of the drivers of this singular epidemiology. We first completed the information available on this human disease in the region by investigating the leptospirosis situation in the Union of the Comoros, a country where no autochthonous transmission has been reported so far. Our results indicate that Comorian populations are exposed to Leptospira, which are antigenically comparable to those detected in the neighbouring island of Mayotte. This finding suggests that the apparent absence of leptospirosis in some islands rather reflects a lack of surveillance. We then investigated the genetic diversity of Leptospira on distinct islands home to distinct endemic animal species that may shed distinct Leptospira lineages and thus at least in part explain the contrasted epidemiology of leptospirosis in the region. Specifically, we describe a high diversity of pathogenic Leptospira within Malagasy bats (Chiroptera) and further show that Leptospira diversity is not structured by geography. Instead, we show that these Leptospira display a strong specificity towards their hosts, which may result from different evolutionary processes including co-speciation and host switching. Using this tight host specificity, we investigated the leptospirosis epidemiology on Mayotte, where we show that the important bacterial diversity reported in clinical cases is due of the presence of several distinct animal species acting as reservoirs, some of which introduced from the neighbouring Madagascar. Altogether, results presented herein combined to data produced by our lab suggest that the epidemiology of leptospirosis in the SWOI results from distinct assemblages of cosmopolitan and autochthonous/endemic Leptospira. Keywords : Leptospirosis, Leptospira, Chiroptera, small mammals, hosts-parasites associations, MAT, MLST, co-phylogeny, Madagascar, Mayotte, Union of Comoros, South-Western Indian Ocean.