O sequenciamento da L. interrogans sorovar Copenhageni e as análises bioinformáticas permitiram a identificação de candidatos vacinais e fatores de virulência. Foram selecionados dois genes, LIC11834 e LIC12253, que foram submetidos a ensaios de presença do DNA genômico e RNA mensageiro em diferentes sorovares de Leptospira. Observamos que o gene LIC12253 foi o mais presente entre os sorovares testados. Os genes foram clonados em vetor de expressão pAE e expressos em E. coli BL21 SI. As proteínas recombinantes foram purificadas e submetidas a ensaio de dicroísmo circular, o qual confirmou que ambas as proteínas estavam estruturadas. Por meio de testes de imunogenicidade em camundongos, ambas as proteínas mostraram-se imunogênicas, apresentando altos títulos de anticorpos, porém não foram capazes de promover resposta imune celular. Em ensaios de localização das proteínas nativas podemos observar a presença destas proteínas na membrana externa de Leptospira. Ensaios de reatividade com soros de pacientes diagnosticados com leptospirose mostraram que há reconhecimento das proteínas por anticorpos presentes nesses soros, sugerindo que as proteínas são expressas durante a infecção. Em ensaios de adesão a componentes de matriz extracelular e componentes do soro e plasma humano, rLIC11834 apresentou ligação à laminina, sendo nomeada de Lsa33, além de ligação ao plasminogênio, ao C4bp e ao fibrinogênio de forma dose-dependente; rLIC12253 apresentou ligação à laminina, sendo chamada de Lsa25, e ao C4bp de forma dose-dependente. Ensaios de desafio demonstraram que as proteínas não apresentam proteção contra infecção letal em hamsters. Assim, acreditamos que estas proteínas multifuncionais possam interagir com proteínas do hospedeiro e ter participação na patogênese da doença. / The genomic sequencing and the advances of bioinformatics analysis allowed the identification of new vaccine candidates and new virulence factors. Therefore, two genes from L. interrogans serovar Copenhageni, LIC11834 and LIC12253 were selected and subjected to assays for the presence of genomic DNA and mRNA in different serovars of Leptospira. These assays found that the gene LIC12253 was the most present among the tested serovars. The genes were then cloned in the expression vector pAE and expressed in E. coli BL21 SI. The recombinant proteins were purified and subjected to circular dichroism, which confirmed that both proteins presented secondary structures. Immunogenicity tests in mice showed that both proteins are immunogenic, with high antibodies titers, but don\'t induce cellular immune response. Localization assays of the native proteins on the leptospiras demonstrated the presence of the proteins on the surface of Leptospira. Reactivity assays with sera of patients diagnosed with leptospirosis showed that the recombinant proteins could be recognized by their antibodies, suggesting that they are expressed during infection. Adhesion assays to the extracellular matrix components, human serum and plasma components, showed that the protein rLIC11834 binds to laminin and was called Lsa33. In addition, Lsa33 interacts to plasminogen, to C4bp and to fibrinogen in a dose-dependent manner. The protein rLIC12253 showed binding to laminina, named Lsa25, and to C4bp in a dose-dependent manner. Challenge assays showed that both recombinant proteins don\'t afford protection against lethal infection in hamsters. Thus, we believe that these multifunctional proteins may interact with host proteins and may play a role in leptospiral pathogenesis.

A leptospirose é uma doença infecto-contagiosa que acomete os animais domésticos, silvestres e o homem, causada por bactérias do gênero Leptospira. No Brasil, a hemorragia pulmonar constitui o principal fator de risco para o óbito com taxas de letalidade para essa forma da doença superior a 50%. Desta maneira, o desenvolvimento de uma vacina preventiva e o melhor entendimento dos mecanismos de lesão envolvidos nos processos de desenvolvimento da leptospirose nos permitirá propor novas terapêuticas a fim de diminuir a alta letalidade associada à doença. Neste trabalho propusemos investigar doze genes de Leptospira interrogans sorovar Copenhageni. A identificação de uma metaloprotease abre caminho para a investigação de uma possível protease envolvida nos processos hemorrágicos para essa importante zoonose. Esperamos, com os resultados apresentados neste trabalho, contribuir com a caracterização da biologia e patogenicidade da leptospirose. / Leptospirosis is an infectious disease that affects domestic animals, wildlife and humans, caused by bacteria of the genus Leptospira. In Brazil, pulmonary hemorrhage is the major risk factor to death with mortality rates for this form of the disease over than 50%. Thus, the development of a preventive vaccine and better understanding of injury mechanisms involved in leptospirosis would allow us to propose new therapies to reduce the high mortality associated with the disease. In the present study we proposed to investigate twelve genes of Leptospira interrogans serovar Copenhageni. The identification of one protease opens the way for the investigation of a possible protease involved in bleeding processes in this important zoonosis. With the results presented here, we expect to contribute to the biology and pathogenicity characterization of leptospirosis.

A leptospirose é uma doença sistêmica, causada por bactérias patogênicas do gênero Leptospira. O desenvolvimento de novas estratégias para prevenir a doença é necessário. As pesquisas atuais têm interesse em identificar antígenos conservados que estão envolvidos nas interações patógeno-hospedeiro. Dois genes de L. interrogans foram selecionados, clonados, expressos e suas respectivas proteínas caracterizadas. Os genes foram amplificados por PCR e clonados no vetor de expressão PAE. As proteínas recombinantes foram purificadas por cromatografia de afinidade ao metal. A proteína rLIC10821 foi capaz de se ligar a laminina, plasminogênio e fibrinogênio. Ambas as proteínas foram localizadas na membrana externa de acordo com as três metodologias utilizadas: imunofluorescência, proteinase K, leptospira intacta. A proteína rLIC10821 que interagiu com o PLG foi capaz de gerar plasmina. Após a interação da proteína rLIC10821 com o fibrinogênio, foi possível identificar uma diminuição de 60% no coágulo de fibrina. / Leptospirosis is a systemic disease caused by pathogenic bacteria of genus Leptospira. The development of new strategies to prevent the disease is needed. Currently research has focused to identify conserved antigens related to the host-pathogen interaction. Two genes of L. interrogans were selected, cloned, expressed and its proteins characterized. The genes were amplified by PCR and cloned into expression vetor pAE. The recombinant proteins were purified by chromatography of metal affinity. The protein rLIC10821 were able to bind to laminin, plasminogen and fibrinogen. Both proteins were localized in the outer membrane according three methodologies: immunofluorescence, proteinase K, intact leptospira. The protein rLIC10821 interacted with PLG was able to generate plasmin. After the interaction of the protein rLIC10821 with fibrinogen, we could identify a decrease of 60% in the fibrin clot.

Leptospirose é uma zoonose mundial que acomete várias espécies de mamíferos, incluindo humanos, causada por espécies de bactérias patogênicas do gênero Leptospira. Possui um quadro de manifestação clínico muito variado, podendo apresentar desde sintomas comuns a outras doenças como febre, calafrios, cefaleia, dores musculares, enjoo, vômitos, diarreia e uma forma mais severa da infecção denominada síndrome de Weill. Seguindo a estratégia de genômica funcional, foram selecionados genes de Leptospira interrogans sorovar Copenhageni, LIC10377, LIC11122, LIC11184 e LIC12287, tendo como critério a predição de localização das proteínas na membrana. Os fragmentos referentes aos genes LIC11122 e LIC12287 foram clonados no vetor pGEM-T Easy e subclonados no vetor de expressão pAE. As proteínas avaliadas interagem com laminina e plasminogênio, de forma dose-dependentes e saturáveis, sugerindo atuam nos processos de patogênese da bactéria. A presença de um fator sigma na superfície celular desempenhando um papel secundário, sugere que a rLIC11122 pode ser uma proteína moonlight. / Leptospirosis is a worldwide zoonosis that affects several species of mammals, including humans, caused by species of pathogenic bacteria of the genus Leptospira. It has a very varied clinical manifestation board and may have symptoms from common tropical diseases such as fever, chills, headache, muscle aches, nausea, vomiting, diarrhea and a severe syndrome of infection known as Weill syndrome. Following the functional genomics strategy, genes were selected from Leptospira interrogans serovar Copenhageni, LIC10377, LIC11122, LIC11184 and LIC12287, with the criterion of the prediction location of proteins in the membrane. The fragments related to genes LIC11122 and LIC12287 were cloned into pGEM-T Easy vector and subcloned into the expression vector pAE. The evaluated proteins interact with laminin and plasminogen, dose-dependent and saturable manner, suggesting participation in bacterial pathogenesis processes. The presence of a sigma factor on the cell surface plays a secondary role, suggests that performs a moonlight rLIC11122 protein.

El documento digital no refiere asesor / Determina la asociación entre la presencia de anticuerpos contra Leptospira sp. y la presentación de problemas reproductivos en borregas de dos localidades de la SAIS Tupac Amaru, durante la campaña reproductiva del 2003. Con este propósito se colectaron muestras de suero de borregas entre 2 a 10 años de edad que tuvieran problemas reproductivos como abortos, vacías simples, vacías dobles (grupo Caso: n = 220) y de borregas sin problemas reproductivos (grupo Control: n = 220) durante la actividad del perneo, para determinar anticuerpos contra Leptospira serovar: pomona, hardjo, canicola, icterohaemorragiae, grippotyphosa, wolfii, ballum y bratislava, mediante la técnica de microaglutinación. La prevalencia de Leptospira sp. En la población estudiada fue de 24.8 ± 4.56%, correspondiendo al grupo Caso 28.64 ± 6.95% y al grupo Control 20.91 ± 5.90%. No se encontró asociación estadística significativa entre las variables presencia de anticuerpos y problemas reproductivos mediante el análisis de Chi cuadrado. Los serovares detectados con mayor frecuencia fueron ballum con 42.2% (46/109) e icterohaemorragiae con 31.19% (34/109). Se detectaron también animales con anticuerpos a más de un serovar de Leptospira. La ausencia de asociación estadística significativa entre las variables mencionadas podría deberse a varios factores como: corta vida de los anticuerpos contra los serovares de Leptospira estudiados, serovares utilizados distintos a los existentes en la zona, época de muestreo, etc. / Tesis

La leptospirose est une zoonose à distribution mondiale dont la prévalence chez le chat varie géographiquement de 4.8% à 35%. Bien que l’exposition féline à Leptospira spp. soit rapportée dans des études sérologiques, les conséquences cliniques de cette maladie chez le chat sont peu connues. Le but principal de cette étude était de comparer le statut sérologique et de porteur (PCR urinaire) de Leptospira spp. entre des chats sains et des chats atteints de maladie rénale (insulte rénale aigue et maladie rénale chronique de stades IIb, III et IV). Une étude préliminaire pour valider la sensibilité et la spécificité analytiques de la PCR de Leptospira spp. réalisée par le Laboratoire de Diagnostic Moléculaire de la FMV sur l’urine de chat a été effectuée. La validation in vitro a démontré que la technique de PCR est efficace pour déterminer la présence de leptospires pathogènes dans l’urine du chat. Dans le cadre de l’étude principale, 251 chats ont été recrutés entre janvier 2010 et mars 2012,. De ceux-ci, 240 ont été inclus et divisés en 2 groupes (chats sains (C=125) et chats atteints de maladie rénale (MR=115) en se basant sur un examen physique ainsi que sur des résultats d’hématologie, de biochimie et d’analyse d’urine. Tous les chats recrutés ont également été examinés sérologiquement par test de micro-agglutination pour la présence d’anticorps contre Leptospira spp. (résultat considéré positif si ≥1 :100) et par PCR pour la présence de Leptospira spp. dans l’urine. Le pourcentage prédit de séropositivité pour Leptospira spp. était significativement plus élevé chez les chats atteints de maladie rénale (13,7%) que chez les chats sains (5%) (p=0,02). Les sérovars impliqués étaient Pomona (n=16), Bratislava (n=8) et Grippotyphosa (n=1). De plus, les chats séropositifs pour Pomona présentaient des titres significativement plus élevés que pour les autres sérovars (p=0,04). L’excrétion de Leptospira spp. a été confirmée par PCR dans l’urine de huit chats. Des 26 chats séropositifs, quatre (C=2, MR=2) se sont également révélés PCR positifs. La prévalence a été plus élevée chez les chats du groupe MR (5.3%; 6/113) lorsque comparée à celle du groupe C (1.6%; 2/125), mais cette différence ne s’est pas révélée statistiquement significative (C=0,9% , MR= 5,5% ; p = 0,09). L’âge, le sexe et le milieu de vie (urbain versus rural) n’ont pas influencé le statut sérologique ou d’excrétion pour Leptospira spp. Le pourcentage prédit de séropositivité était significativement plus élevée chez les chasseurs (p<0.01) et pendant les mois de juin à août (p=0.02). La présence d’un autre chat à la maison a également significativement augmenté ce pourcentage (p<0.01), mais la présence d’un chien ne l’a pas influencé. Lors de l’évaluation du PCR par le modèle GGE, seules les variables « contact avec raton laveur » et « contact avec mouffettes » sont ressorties statistiquement significatives (p≤0.03). Le rôle que joue Leptospira spp. comme agent étiologique de maladie rénale chez le chat demeure incertain. Toutefois, la différence significative de statut sérologique entre les chats sains et les chats atteints de maladie rénale suggère que la leptospirose pourrait être une cause sous-diagnostiquée de maladie rénale chez cette espèce. Dans cette étude, plusieurs porteurs asymptomatiques ont été identifiés, ce qui suggère que l’espèce féline puisse être un acteur sous-estimé dans la transmission de la bactérie aux humains. / Leptospirosis is a globally widespread zoonosis, with prevalence in cats varying from 8.8% to 35% depending on geographical localization. Although serologic evidence of feline exposure exists, clinical disease is rarely reported. This study aimed to compare seropositivity and urinary PCR status for Leptospira spp. between healthy cats (H) and cats with kidney disease (KD: acute kidney injury (AKI) and chronic KD stages IIb, III and IV). The analytical sensitivity and specificity of the PCR performed by the Laboratoire de Diagnostic Moléculaire of the FMV were evaluated. In vitro validation showed that the PCR technique is effective for determining the presence of pathogenic leptospires in the urine of cats. A total of 251 cats were recruited, from which 240 cats were enrolled. Cats were enrolled from January 2010 to March 2012 and divided into two groups, H (n=125) and KD (n=115), based on complete blood count, serum biochemistry profile and urinalysis. Leptospira spp. serology by microscopic agglutination test (titers ≥ 1:100 considered positive) and urinary PCR were performed in all cats. Predicted seropositivity for Leptospira spp. was statistically different between groups; being 5% and 13,7% in the H and KD groups respectively (p=0.02). Serovars involved were Pomona (n=16), Bratislava (n=8) and Grippotyphosa (n=1), with titers being significantly higher for Pomona (p=0.04). The excretory status was confirmed by a positive urine PCR in eight cats. Of the 26 seropositive cats, four (H=2, KD=2) were also PCR positive. The prevalence of PCR positive cats was higher in the KD group (5.3%; 6/113) compared with the H group (1.6%; 2/125), but the difference between groups did not reach statistical significance (0.9% in H, 5.5% in KD; p=0.09). Age, sex and rural versus urban environment did not influence serologic or PCR status for Leptospira spp. Predicted seropositivity was greater between June and August (p =0.02) and in known hunters (p<0.01). The presence of another cat at home also increased significantly the predicted seropositivity (p<0.01), although the presence of a dog did not. When evaluating the PCR status of cats in the GEE model for individually tested variables, only the variables “contact with raccoons” and “contact with skunks” were statistically significant (P ≤ .03). Although the precise role of Leptospira spp. as an etiologic agent of feline KD remains unclear, the significant difference in the serologic status found between H and KD cats suggests that it may be an under-diagnosed cause of KD in cats. Several asymptomatic carriers were identified, suggesting that cats could be underestimated players in the transmission of the bacteria to humans.

La leptospirose est une zoonose à distribution mondiale dont la prévalence chez le chat varie géographiquement de 4.8% à 35%. Bien que l’exposition féline à Leptospira spp. soit rapportée dans des études sérologiques, les conséquences cliniques de cette maladie chez le chat sont peu connues. Le but principal de cette étude était de comparer le statut sérologique et de porteur (PCR urinaire) de Leptospira spp. entre des chats sains et des chats atteints de maladie rénale (insulte rénale aigue et maladie rénale chronique de stades IIb, III et IV). Une étude préliminaire pour valider la sensibilité et la spécificité analytiques de la PCR de Leptospira spp. réalisée par le Laboratoire de Diagnostic Moléculaire de la FMV sur l’urine de chat a été effectuée. La validation in vitro a démontré que la technique de PCR est efficace pour déterminer la présence de leptospires pathogènes dans l’urine du chat. Dans le cadre de l’étude principale, 251 chats ont été recrutés entre janvier 2010 et mars 2012,. De ceux-ci, 240 ont été inclus et divisés en 2 groupes (chats sains (C=125) et chats atteints de maladie rénale (MR=115) en se basant sur un examen physique ainsi que sur des résultats d’hématologie, de biochimie et d’analyse d’urine. Tous les chats recrutés ont également été examinés sérologiquement par test de micro-agglutination pour la présence d’anticorps contre Leptospira spp. (résultat considéré positif si ≥1 :100) et par PCR pour la présence de Leptospira spp. dans l’urine. Le pourcentage prédit de séropositivité pour Leptospira spp. était significativement plus élevé chez les chats atteints de maladie rénale (13,7%) que chez les chats sains (5%) (p=0,02). Les sérovars impliqués étaient Pomona (n=16), Bratislava (n=8) et Grippotyphosa (n=1). De plus, les chats séropositifs pour Pomona présentaient des titres significativement plus élevés que pour les autres sérovars (p=0,04). L’excrétion de Leptospira spp. a été confirmée par PCR dans l’urine de huit chats. Des 26 chats séropositifs, quatre (C=2, MR=2) se sont également révélés PCR positifs. La prévalence a été plus élevée chez les chats du groupe MR (5.3%; 6/113) lorsque comparée à celle du groupe C (1.6%; 2/125), mais cette différence ne s’est pas révélée statistiquement significative (C=0,9% , MR= 5,5% ; p = 0,09). L’âge, le sexe et le milieu de vie (urbain versus rural) n’ont pas influencé le statut sérologique ou d’excrétion pour Leptospira spp. Le pourcentage prédit de séropositivité était significativement plus élevée chez les chasseurs (p<0.01) et pendant les mois de juin à août (p=0.02). La présence d’un autre chat à la maison a également significativement augmenté ce pourcentage (p<0.01), mais la présence d’un chien ne l’a pas influencé. Lors de l’évaluation du PCR par le modèle GGE, seules les variables « contact avec raton laveur » et « contact avec mouffettes » sont ressorties statistiquement significatives (p≤0.03). Le rôle que joue Leptospira spp. comme agent étiologique de maladie rénale chez le chat demeure incertain. Toutefois, la différence significative de statut sérologique entre les chats sains et les chats atteints de maladie rénale suggère que la leptospirose pourrait être une cause sous-diagnostiquée de maladie rénale chez cette espèce. Dans cette étude, plusieurs porteurs asymptomatiques ont été identifiés, ce qui suggère que l’espèce féline puisse être un acteur sous-estimé dans la transmission de la bactérie aux humains. / Leptospirosis is a globally widespread zoonosis, with prevalence in cats varying from 8.8% to 35% depending on geographical localization. Although serologic evidence of feline exposure exists, clinical disease is rarely reported. This study aimed to compare seropositivity and urinary PCR status for Leptospira spp. between healthy cats (H) and cats with kidney disease (KD: acute kidney injury (AKI) and chronic KD stages IIb, III and IV). The analytical sensitivity and specificity of the PCR performed by the Laboratoire de Diagnostic Moléculaire of the FMV were evaluated. In vitro validation showed that the PCR technique is effective for determining the presence of pathogenic leptospires in the urine of cats. A total of 251 cats were recruited, from which 240 cats were enrolled. Cats were enrolled from January 2010 to March 2012 and divided into two groups, H (n=125) and KD (n=115), based on complete blood count, serum biochemistry profile and urinalysis. Leptospira spp. serology by microscopic agglutination test (titers ≥ 1:100 considered positive) and urinary PCR were performed in all cats. Predicted seropositivity for Leptospira spp. was statistically different between groups; being 5% and 13,7% in the H and KD groups respectively (p=0.02). Serovars involved were Pomona (n=16), Bratislava (n=8) and Grippotyphosa (n=1), with titers being significantly higher for Pomona (p=0.04). The excretory status was confirmed by a positive urine PCR in eight cats. Of the 26 seropositive cats, four (H=2, KD=2) were also PCR positive. The prevalence of PCR positive cats was higher in the KD group (5.3%; 6/113) compared with the H group (1.6%; 2/125), but the difference between groups did not reach statistical significance (0.9% in H, 5.5% in KD; p=0.09). Age, sex and rural versus urban environment did not influence serologic or PCR status for Leptospira spp. Predicted seropositivity was greater between June and August (p =0.02) and in known hunters (p<0.01). The presence of another cat at home also increased significantly the predicted seropositivity (p<0.01), although the presence of a dog did not. When evaluating the PCR status of cats in the GEE model for individually tested variables, only the variables “contact with raccoons” and “contact with skunks” were statistically significant (P ≤ .03). Although the precise role of Leptospira spp. as an etiologic agent of feline KD remains unclear, the significant difference in the serologic status found between H and KD cats suggests that it may be an under-diagnosed cause of KD in cats. Several asymptomatic carriers were identified, suggesting that cats could be underestimated players in the transmission of the bacteria to humans.

Memoria para optar al Título Profesional de Médico Veterinario. / La leptospirosis es una zoonosis de distribución mundial causada por bacterias de tipo espiroqueta del género Leptospira. La transmisión de este patógeno comúnmente se produce por contacto directo o indirecto con orina de animales infectados. Los animales de vida silvestre juegan un papel importante en la trasmisión de la bacteria, siendo los roedores el principal reservorio silvestre. Las fuentes de agua también favorecen la diseminación pudiendo permanecer en ella por largos períodos de tiempo. Con el fin de identificar Leptospiras patógenas y su relación de seropositividad con variables biológicas y de manejo de perros en la comuna de La Pintana se tomaron muestras de sangre de 119 caninos domésticos. Los sueros fueron analizados mediante la técnica de microMAT para nueve serovares. Además se realizó una encuesta a los propietarios para determinar manejos e información individual de cada perro. Se detectaron Leptospiras patógenas en 15 de 119 individuos, siendo el serovar Canicola el más frecuente. Las variables vacunación séxtuple, grado de supervisión al pasear, consumo de agua potable y contacto con roedores, presentaron una relación significativa con seropositividad, pudiendo ser los roedores una fuente de diseminación en las zonas semiurbana y la falta de conocimiento de los propietario con respecto a la Tenencia Responsable de sus mascotas los principales factores que favorecen la infección de los cánidos. / Leptospirosis is a zoonosis with global distribution caused by bacteria of the spirochete type of the genus Leptospira. The transmission of this pathogen is produced by direct or indirect contact with urine of infected animals. Wildlife animals play an important role in the transmission of the bacteria, with rodents being the main wild reservoir. Water sources also favor dissemination of the bacteria, which can remain in it for long periods of time. To identify pathogenic Leptospiras and their relationship between seropositivity with biological variables and the management of dogs in La Pintana commune, blood samples were taken from domestic canines. To assess the presence of nine serotypes of Leptospira, the microMAT technic was used. A survey was applied to owners to collect information on management and individual factors of the dogs. Pathogenic Leptospira were detected in 15 of 119 individuals, with Canicola being the most frequent serovar. The variables vaccination, degree of supervision during walks, consumption of drinking water and contact with rodents, had a significant association with seropositivity, rodents being a source of dissemination in the semi-urban areas and the lack of knowledge of the owners regarding the responsible ownership of pets being the main factors that favor the infection of canids. / Financiamiento: Proyecto U-Inicia No. 121017019102049.

Orientador: Helio Langoni / Resumo: O interesse pelo estudo das zoonoses, enfermidades comuns aos humanos e aos animais, aumentou acentuadamente nos últimos anos, sugerindo maior integração, conhecimento e relacionamento entre profissionais da área da saúde. Dentre estas zoonoses a brucelose, a toxoplasmose, a leptospirose e a doença de Chagas (DC) têm elevada importância veterinária e para a saúde pública, pela gravidade e potencial letalidade com que se apresentam em humanos. Atenção especial deve ser dada às pessoas vivendo com HIV/aids (PVHA), pois a imunossupressão à que estão sujeitas pode agravar ainda mais o seu quadro clínico, fato que gera necessidade de diagnósticos mais eficientes para essas infecções. Com o objetivo de determinar o perfil sorológico destas zoonoses em PVHA, atendidos no Serviço de Ambulatório Especializado de Infectologia “Domingos Alves Meira” (SAEI-DAM), entidade pertencente ao complexo de assistência da Faculdade de Medicina de Botucatu – UNESP, bem como relacionar aos hábitos, comportamentos e fatores de risco de exposição a essas doenças, além da obtenção de dados referentes ao grau de conhecimento em relação às zoonoses estudadas. Foram analisadas 236 amostras de soro desses pacientes para a presença de anticorpos anti- Brucella abortus, que mostraram-se negativas para todas as amostras. Para leptospirose quatro (1,69%) amostras foram positivas, para a toxoplasmose 154 (65%) amostras positivas, sendo 146 (94,8%) somente para anticorpos da classe IgG, dois (1,3%) somente para ... (Resumo completo, clicar acesso eletrônico abaixo) / Doutor

A criação industrial de avestruzes no Brasil tem crescido nos últimos anos, mas avestruzes podem ser reservatórios de agentes com potencial zoonótico, como é o caso do vírus da influenza, rotavírus, vírus da encefalomielite eqüina do leste (EEEV) e do oeste (WEEV), salmonela, leptospira e micoplasma. O objetivo deste trabalho foi detectar rotavírus nas fezes de filhotes, anticorpos contra rotavírus, EEEV e WEEV, influenza A , leptospira, salmonela e micoplasma a partir do soro de reprodutores e efetuar o isolamento de Salmonella sp. a partir de de suabe cloacal. Para a detecção de rotavírus nas fezes utilizaram-se as técnicas de PAGE, isolamento viral e genotipagem pela RT-PCR. As técnicas de contraimunoeletroosmoforese, soroneutralização em cultura de células, inibição da hemaglutinação e aglutinação em placa foram utilizadas nos levantamentos sorológicos e a cultura bacteriológica foi utilizada para detecção de Salmonella spp. em suabes de cloaca. Rotavírus do grupo A com genotipos G[6], G[10], P[1] e P[7] foram detectados nas fezes de avestruzes e anticorpos anti-rotavírus em 10 amostras (10/182) de soros colhidos de avestruzes reprodutores. Foram detectados anticorpos para vários sorovares de Leptospira spp. (19/128), Salmonella pullorum (17/182), Mycoplasma gallisepticum (17/182) e Mycoplasma synoviae (47/182). Não foram detectados anticorpos anti-EEEV e WEEV e anti-vírus da Influenza A H3, bem como amostras de Salmonella spp. em suabes de cloaca. Estes resultados permitem sugerir o papel do avestruz na cadeia epidemiológica das rotaviroses e da leptospirose e dão bases para o aprimoramento sanitário do plantel brasileiro desta ave. / The industrial ostrich breeding in Brazil has grown in the last few years, but ostrich may be reservoirs of potentially zoonotic agents such as influenzavirus, rotavirus, eastern equine encephalitis virus (EEEV), western equine encephalitis virus (WEEV), Salmonella, Leptospira and Mycoplasma. The aim of the present study was to detect rotavirus in fecal samples of young ostriches, antibodies against rotavirus, EEEV and WEEV, influenza A , Leptospira, Sallmonela and Mycoplasma in sera from breeders and carry out Salmonella isolation in cloacal swabs. For the rotavirus detection, PAGE, viral isolation and genotyping by RT-PCR were used. Counterimmunoelectroosmophoresis, virus neutralisation assay in cell culture, hemagglutination inhibition and plate agglutination were used in the serological surveys and bacteriological culture was used to detetc Salmonella spp. in the cloacal swabs. Group A rotavirus with genotypes G[6], G[10], P[1] and P[7] was detected in the fecal samples and anti-rotavirus antibodies were detected in ten samples (10/182) from breeder ostriches. Antibodies to different serovars of Leptospira spp. (19/128), Salmonella pullorum (17/182), Mycoplasma gallisepticum (17/182) and Mycoplasma synoviae (47/182) were detected. Antibodies against EEEV, WEEV and influenzavirus A H3 were not detected, as well as Salmonella spp. in swab samples. These results allow one to suggest a role to ostriches in the epidemiological chain of rotavirus diseases and leptospirosis and give basis to the improvement of the sanitary condition of the Brazilian flocks.