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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
191

Synthèse de composés polyspiranniques par oxydation phénolique -<br />Synthèse totale des aculéatines et d'analogues antipaludiques

Peuchmaur, Marine 12 October 2007 (has links) (PDF)
Les aculéatines sont des molécules naturelles, biologiquement actives, isolées de Amomum aculeatum, une plante utilisée en médecine traditionnelle (Papouasie Nouvelle Guinée) pour le traitement des fièvres et du paludisme. L'approche synthétique développée dans le cadre de la synthèse totale énantiosélective de ces molécules utilise comme réaction clef une étape d'oxydation phénolique associée à une spiroannélation. L'étude de la réaction d'aldolisation diastéréosélective intervenant dans cette approche nous a permis d'étendre le champ d'application des inductions 1,3 anti (PMB) aux substrats non encombrés, mais également de mettre en évidence une induction 1,3 de type syn (Tr), rarement décrite dans la littérature.<br />A l'issue de la mise au point de cette synthèse deux thématiques ont été développées.<br />Un premier axe de recherche a visé l'étude de l'étape d'oxydation phénolique et, plus particulièrement, l'étude du piégeage du cation phénoxonium par différents hétéroatomes hybridés sp2 : l'oxygène d'une fonction cétone dans le cas des aculéatines, l'oxygène d'une fonction lactame lors d'une tentative de synthèse de la spiroleucettadine, ou encore l'azote d'une oxazoline au travers d'une approche synthétique d'un analogue des TAN.<br />D'un point de vue biologique, la synthèse d'analogues des aculéatines a rendu possible l'évaluation des paramètres importants intervenant dans trois activités biologiques (antipaludique, antitoxoplasmique, herbicide) possédant comme cible commune les plastes. Certaines caractéristiques structurales ont pu être dégagées des études de relation structure-activité. De même, deux molécules actives sur P. falciparum (CI50 de 80 et 90 nM) sont en cours d'étude in vivo.
192

Synthèses et évaluations de nouveaux composés antipaludiques et antitoxoplasmoses

Traore, Mariam 07 December 2012 (has links) (PDF)
Le paludisme est la maladie parasitaire la plus meurtrière qui touche 3,3 milliards de personne (la moitié de la population mondiale). En 2010, le nombre des décès dus au paludisme a été estimé à 655 milles, dont 91% des cas concerne le continent Africain. Ce sont les femmes enceintes et les enfants de moins de 5 ans (86% des décès) qui sont les plus vulnérables. Pour les pays africains le paludisme est non seulement une maladie due à la pauvreté, mais il est également la cause de la pauvreté. Face à ce fléau qui fait beaucoup de ravage dans les pays pauvres, il est urgent de trouver de nouveaux traitements efficaces et accessibles aux populations concernées. D'autre part, la toxoplasmose est une maladie parasitaire qui touche principalement les pays industrialisés (50% de la population française). Cette parasitose souvent bénigne chez les personnes immunocompétentes, est qualifiée de maladie opportuniste, car elle peut s'avérer dangereuse chez les femmes enceintes et chez les patients immunodéprimés (personnes atteintes du SIDA ou ayant subi des transplantations d'organe). Les parasites responsables de paludisme (P. falciparum) et de la toxoplasmose (T. gondii) appartiennent à la grande famille des apicomplexes, qui sont des organismes unicellulaires. Afin de concevoir de nouveaux composés antipaludiques et antitoxoplasmoses, nous nous sommes inspirés de produits d'origine naturelle qui sont les aculéatines et les cyclotétrapeptides analogues de FR235222. Afin d'optimiser les propriétés antiparasitaires de ces produits naturels, nous avons développé des méthodes de synthèse rapides en peu d'étapes. Des nouveaux analogues des aculéatines ont été obtenus en utilisant une stratégie de réactions en cascade et tandem d'oxydation phénolique induites par les réactifs à l'iode hypervalent (III). Des réactions de chimie radicalaire avec les xanthates et thioesters ont permis d'obtenir une grande diversité de cyclotétrapeptides analogues de FR235222. Les activités antiparasitaires des différents analogues obtenus ont été évaluées et certains se sont révélés actifs de l'ordre du nanomolaire.
193

Perfil sorológico em candidatos à doação de órgãos sólidos no estado de Santa Catarina no período de 2001 a 2007

Baumel, Rosana Marquardt 24 October 2012 (has links)
Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências da Saúde, Programa de Pós-graduação em Saúde Pública, Florianópolis, 2009 / Made available in DSpace on 2012-10-24T15:36:06Z (GMT). No. of bitstreams: 0 / Objetivo: O objetivo deste estudo foi analisar o perfil sorológico dos marcadores de doenças entre os candidatos à doação de órgãos sólidos no Estado de Santa Catarina no período de janeiro de 2001 a dezembro de 2007. Métodos: Foi conduzido estudo epidemiológico, transversal baseado em dados secundários. A população estudada foi composta por 1424 candidatos à doação de órgãos sólidos, no Estado de Santa Catarina. O perfil sorológico das doenças da população do estudo foi realizado pelo Centro de Hematologia e Hemoterapia de Santa Catarina, HEMOSC e os dados sócio-econômicos foram captados pela Central de Notificação, Captação e Distribuição de Órgãos de Santa Catarina, CNCDO/SC. As variáveis analisadas neste estudo foram os resultados das sorologias realizadas nos candidatos à doação de órgãos sólidos, sexo, idade e ano do exame. Resultados: O estudo constitui-se de 1424 dados das sorologias de candidatos à doação de órgãos sólidos, 551 (38,7%) do sexo feminino e 873 (61,3%), do sexo masculino. Os marcadores sorológicos em candidatos à doação de órgãos sólidos foram: anti-HBc=21,7%, HBsAg= 3,9%, anti-HCV=1,3, doença de Chagas=0,3%, sífilis=0,9%, HTLV I/II =0,3%, HIV I/II=0,5%, toxo IgG=38,2%, toxo IgM=2,3%, CMV IgG=46,2% e CMV IgM=2,6%. A soroprevalência de CMV IgM foi maior nas mulheres (p<0,01). Os outros marcadores não apresentaram diferença estatisticamente significativa entre os sexos. Conclusões: Houve maior prevalência do sexo masculino entre os candidatos à doação de órgãos e aumento na faixa etária acima de 60 anos. A exposição ao vírus da hepatite B é comum e crescente em Santa Catarina ao longo do tempo sendo o marcador HBsAg maior em homens. As soroprevalências de HCV, HIV I/II, HTLV I/II e doença de Chagas foram baixas sem alterações ao longo do período estudado. As infecções por citomegalovírus e toxoplasmose se mostraram elevadas, sendo a infecção aguda pouco comum. Citomegalovírus foi mais freqüente em mulheres e nos mais jovens. / Objective: This study examined the profile of serological markers of blood-borne diseases among the candidates for donation of solid organs. Methods: We conducted an epidemiological survey based on secondary data. The study population comprised 1424 candidates for donation of solid organs in the Santa Catarina State from January 2001 to December 2007. Serological analyses were conducted by the Center for Hematology and Hemotherapy of the Santa Catarina State (HEMOSC). Demographic data were collected by the Centre for Organs donation of the state (CNCDO/SC). The variables analyzed included serological tests results in the candidates for solid organ donation, as well as their sex, age and year of examination. Results: Among 1424 subjects examined, there were 551 (38.7%) females and 873 (61.3%) males. Seroprevalence of disease markers was as following: 21.7% with anti-HBc, 3,9% with HBsAg, 1,3% with anti-HCV, 0,3% with Chagas disease, 0,9% with VDRL marker for syphilis, 0,3% with HTLV 1 or 2, 0,5% with HIV 1 or 2, 38.2% with toxoplasmosis IgG, 2,3% with toxoplasmosis IgM, 46,2% with CMV IgG and 2,6% with CMV IgM. The seroprevalence of CMV IgM was higher in women (p <0.01). The other markers showed no statistically significant difference between the sexes. Conclusions: There was a higher percentage of males among the candidates for donation of organs. During the period analyzed, an increase in the percentage of candidates for donation over 60 years of age and of HBsAg-positive men was observed. The seroprevalences of HCV, HIV, HTLV and Chagas disease were low and with no change over the period studied. The infections by cytomegalovirus and toxoplasmosis were highly prevalent but rarely in the acute phase. Cytomegalovirus was more frequent in women and younger people.
194

Stratégie vaccinale contre la toxoplasmose : ciblage d'un antigène paratisaire aux cellules dendritiques par un fragment d'anticorps de type scFv / Vaccine strategy against toxoplasmosis : parasite antigen targeting to dendritic cells by scFv fragment antibody (single chain antibody fragment)

Lakhrif, Zineb 11 December 2015 (has links)
La toxoplasmose est un problème majeur de Santé Publique et notamment en médecine humaine. Le développement de vaccins est donc d’une grande priorité. L’efficacité de la stratégie vaccinale contre Toxoplasma gondii dépend de l’induction des réponses immunitaires muqueuse et systémique Th1. Les cellules dendritiques (CDs) ont un rôle essentiel dans l'orchestration de l’immunité innée et l’induction de l’immunité adaptative spécifique à Toxoplasma gondii. Dans cette étude, nous explorons une stratégie de vaccination originale qui consiste en l’administration par voie systémique et muqueuse de protéines de fusion capables de cibler l’antigène de surface SAG1 aux CDs, en utilisant un fragment d'anticorps de type scFv dirigé contre le récepteur d'endocytose DEC205. Nos résultats montrent que le ciblage de SAG1 aux DCs par le fragment scFv via la voie intranasale et sous-cutanée, réduit dramatiquement la charge parasitaire cérébrale par rapport à l’antigène non ciblé et est plus efficace que l’immunisation par la voie intranasale ou la voie sous-cutanée seule. Le ciblage des DCs potentialise la réponse immunitaire vers un profil Th1 par la production d’IFN-γ, d’IL-2, d’IgG2a sériques, tout en favorisant la production IgA muqueuses spécifiques. Parallèlement, nous avons montré qu’il était possible de conférer une reconnaissance par la protéine L à toutes les chaînes légères kappa, ce qui permettra dans l’avenir de purifier plus efficacement les antigènes ciblés par chromatographie d’affinité avec la protéine L. / Toxoplasmosis is a major public health problem and the development of a human vaccine is of high priority. Efficient vaccination against Toxoplasma gondii (T. gondii) requires both a mucosal and systemic Th1 immune response. Moreover, dendritic cells (DCs) play a critical role in orchestrating the innate immune functions and driving specific adaptive immunity to T. gondii. In this study, we explore an original vaccination strategy that combines administration via mucosal and systemic routes of fusion proteins able to target the major T. gondii surface antigen SAG1 to DCs using an antibody fragment scFv directed against DEC205 endocytic receptor. Our results show that SAG1 targeting to DCs by scFv via intranasal and subcutaneous administration improved protection against chronic T. gondii infection. A marked reduction in brain parasite burden is observed when compared with the intranasal or the subcutaneous route alone. DC targeting improved both local and systemic humoral and cellular immune responses and potentiated more specifically the Th1 response profile by more efficient production of IFN-γ, IL-2, IgG2a and nasal IgA. In parallel, we found a strategy to confer PpL recognition to all kappa chains. Therefore, affinity chromatography on protein L (PpL) matrix could be used to get easily highly purified targeted proteins.
195

Detecção molecular de coccídios da familia sarcocystidae em amostras teciduais de pequenos felídeos neotropicais do Rio Grande do Sul / Molecular detection of coccidia family Sarcocystidae in tissues samples of small Neotropical wildlife felids of Rio Grande do Sul

Cañón-Franco, William Alberto January 2013 (has links)
Poucos estudos quantificam o risco relativo da saúde humana na transmissão spillover de doenças zoonóticas de populações de animais silvestres, estudos cruciais na compreensão da história natural das zoonoses. Coccídios, em particular os da família Sarcocystidae, são importantes agentes transmissíveis na interface homens - animais domésticos e silvestres. O diagnóstico da coccidiose é prejudicado pela limitada disponibilidade de amostras resultantes de populações animais de espécies em risco de extinção. O objetivo deste estudo foi detectar através da amplificação do locus ITS-1, protozoários das subfamílias Sarcocystinae e Toxoplasmatinae em amostras teciduais de Puma yagouaroundi, Leopardus geoffroyi, L. tigrinus, L. wiedii, L. colocolo e L. pardalis, depositados em coleções biológicas do Rio Grande do Sul, Brasil. Um objetivo adicional foi a obtenção de informações que permitissem avaliar o papel epidemiológico dos protozoários no ciclo silvestre dos parasitas e seu possível impacto sobre as populações de animais silvestres e na saúde pública. Noventa pequenos felídeos neotropicais de vida livre, representando seis espécies, foram amostrados. Destes, 31 felídeos (34,4%), de todas as seis espécies, foram positivos para Toxoplasma gondii e DNA foi detectado em 63 das 433 (14,6%) amostras de tecidos primários coletados a partir de língua (28,6%), cérebro (18,6%), músculo esquelético (17,1%), musculatura ocular (13,6%), globo ocular (13,6%), coração (11,1%), diafragma (5,4%) e humor vítreo (4,5%). Doze amostras primárias positivas ao T. gondii foram genotipadas com os marcadores moleculares SAG1, SAG2, SAG3, BTUB, GRA6, c22- 8, c29-2, L358, PK1, Apico e CS3 e a técnica multilocus PCR-RFLP, a amostra Py#36m foi totalmente caracterizada como do tipo I com alelo II no BTUB e um novo genótipo atípico Py#21M, ambos isolados de Puma yagouaroundi e nunca descrito no Brasil. Nove outras amostras tiveram caracterização parcial. Treze dos 90 felídeos foram positivos para Sarcocystis spp. (14,4%) e outros 18 felídeos, representando cinco espécies albergaram S. felis-like [Py (#75m, #83m, #35m, #20li, #55li), Lg (#80m, #70m, #88m, #71li, #67mOi), Lt (#19m, #48m, #89m, #84m), Lw (#12, #73d) e Lc (#82m, #76m)]. Um único felino de L. pardalis foi negativo. DNA do parasita foi detectado em 11,8% dos tecidos examinados (51/433): musculatura esquelética (26,5%), língua (23,2%), musculatura ocular (13,6%), diafragma (10,7%), cérebro (2,3%), coração (1,6%) e globo ocular (4,5%), nenhuma das 44 amostras de humor vítreo foi positiva. Esta é a primeira detecção e caracterização genética de T. gondii e de S. felislike em felídeos silvestres brasileiros de vida livre, demonstrando a presença destes agentes no ciclo silvestre e, a potencial transmissibilidade ao homem e a outros animais domésticos e silvestres. O uso de amostras de tecidos de animais silvestres depositados em coleções biológicas para estudos epidemiológicos de doenças monstraram serem de grande utilidade. / Few studies have quantified the relative risk of human health from spillover of zoonotic diseases from populations of wild animals; these studies are crucial for understanding the natural history of zoonoses. Coccidia, particularly from the family Sarcocystidae, are important transmissible agents at the interface of man and domestic and wild animals. The diagnosis of Coccidiosis is hampered by the limited availability of samples resulting from protection of natural populations of the species at risk of extinction. The aim of this study was to detected, by amplification of ITS-1 locus, protozoa from the subfamilies Sarcocystinae and Toxoplasmatinae in tissue samples from Puma yagouaroundi, Leopardus geoffroyi, L. tigrinus, L. wiedii, L. colocolo and L. pardalis, deposited in biological collections of the State of Rio Grande do Sul, Brazil. An additional aim was to obtain information that would enable assessment of the epidemiological role of the protozoa in the sylvatic cycle of the parasite, and its possible impact on wildlife populations and public health. Ninety free-living small wild felines, representing 6 species, were sampled. Of these, 31 felids (34.4%) of all six species were positive for T. gondii and DNA was detected in 63 of 433 (14.6%) primary tissue samples collected from the tongue (28.6%), brain (18.6%), skeletal muscle (17.1%), ocular muscles (13.6%), eye (13.6%), heart (11.1%), diaphragm (5.4%) and vitreous humor (4.5%). Twelve primary samples positive for T. gondii were genotyped with molecular markers SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and apical CS3. Using the multilocus PCR-RFLP technique, sample Py#36m was fully genotyped as Type I with allele II in locus BTUB, and a new atypical Py#21M, both isolates from Puma yagouaroundi and never described in Brazil. Nine other samples had a partial characterization. Thirteen of the 90 felids were positive for Sarcocystis spp. (14.4%) and another 18 felids, representing 5 species, harbored S. felis-like organisms [Py (#75m, #83m, #35m, #20li, #55li), Lg (#80m, #70m, #88m, #71li, #67mOi), Lt (#19m, #48m, #89m, #84m), Lw (#12, #73d) and Lc (#82m, #76m)]. A single felid of L. pardalis was negative. Parasite DNA was detected in 11.8% (51/433) of the tissues examined: muscle skeletal (26.5%), tongue (23.2%), ocular muscles (13.6%), diaphragm (10.7 %), brain (2.3%), heart (1.6%) and eye (4.5%); none of the 44 samples of vitreous humor was positive. This is the first description of the detection and genetic characterization of T. gondii and S. felis-like in free-living Brazilians wild felids, demonstrating the presence of these agents in the sylvatic cycle, and the potential transmition to humans and other domestic and wild animals. The use of tissue samples from wild animals deposited in biological collections for epidemiological studies of diseases demonstrated to be of great utility.
196

Estudos de soroprevalência de toxoplasmose em gestantes atendidas na rede Municipal de Saúde de Vitória, ES

Areal, Kelly Rose 07 May 2007 (has links)
Made available in DSpace on 2016-12-23T13:56:01Z (GMT). No. of bitstreams: 1 Tese_kelly_pdf.pdf: 1882941 bytes, checksum: dbb7b527acb39029842a16c46fd45c74 (MD5) Previous issue date: 2007-05-07 / Introduction: Toxoplasmosis is prevalent in Brazil. The infection during pregnancy can affect the fetus and cause sequelae in the child. Objectives: To determine seroprevalence of toxoplasmosis in pregnant women attending primary health cares in Vitória Municipality and evaluate correlate factors for the infection. Methods: A cross-sectional study, performed from January to December 2006, in 1153 pregnant women attending antenatal care in public clinics from the six health areas. A face-to-face interview with sócio-demographic, behaviors and clinics was performed; and serology for IgG, IgM and avidity of IgG by quimiluminescence (Diasorin) and eletroquimioluminescence (Biolab-Merrieux). Results: Prevalence of toxoplasmosis was 73.5% (CI 95% 70.95%-76.05%) and 1.3% (CI 95% 0.65%-1.95%) of acute infection. Considering avidity of IgG, prevalence of acute infection was 1.1% (CI 95% 0.5%-1.7%). A total of 26.5% (CI 95% 23.9%-29.0%) pregnant women were susceptive to toxoplasmosis and 72.2% (CI 95% 69.6%-74.8%) immunes. IgG antibodies were independent associated to buying meat in free markets [1.78 (CI 95% 1.02-3.13), and IgM antibodies during pregnancy were associated to lower education (up to four years) [5.30 (CI95% 1.67-16.83)]. Conclusions: These results corroborate the importance of earlier antenatal care and the inclusion of avidity test in toxoplasmosis diagnosis. It is requested a better sanitary control concerning the commerce of meat without registration. / Introdução: A toxoplasmose é de alta prevalência no Brasil. A infecção durante a gravidez pode resultar em doença fetal com graves seqüelas para a criança. Objetivo: Determinar a soroprevalência de toxoplasmose em gestantes atendidas nas Unidades de Saúde do Município de Vitória e avaliar fatores correlatos com a infecção. Métodos: Estudo de corte transversal, realizado de janeiro a dezembro de 2006, em 1153 gestantes atendidas nas Unidades de Saúde das seis regiões de saúde. Entrevista face-a-face contendo dados sócio-demográficos, epidemiológicos e clínicos foi realizada e sorologias para pesquisa de IgG, IgM e avidez de IgG pelo método de quimiluminescência (Diasorin) e eletroquimioluminescência (Biolab-Merrieux). Resultados: A prevalência da infecção foi de 73,5% (IC 95% 70,95%-6,05%) para IgG e 1,3% (IC 95% 0,65%-1,95%) para IgM. Quando se considerou a avidez de IgG, a prevalência de infecção aguda foi de 1,1% (IC 95% 0,5%-1,7%). Um total de 26,5% (IC 95% 23,9%-29,0%) gestante era susceptível à toxoplasmose e 72,2% (IC 95% 69,6%-74,8%) imunes. A presença de anticorpos IgG esteve independentemente associada à aquisição de carne em feiras livres [1,78 (IC95% 1,02-3,13). Já a presença de anticorpos IgM durante a gravidez apresentou associação com uma menor escolaridade (até quatro anos de estudo) [5,30 (IC95% 1,67-16,83)]. Conclusão: Estes resultados corroboram a importância da adesão precoce ao pré-natal com a inclusão do ensaio de avidez de IgG no diagnóstico da toxoplasmose. É necessário haver um maior rigor nas exigências sanitárias, no que diz respeito ao comércio de carnes sem registro.
197

Frequ?ncia da infec??o por Toxoplasma gondii em galinhas caipira e frangos de corte em regi?es dos Estados do Rio Grande do Norte e Para?ba

Santos, Maria Cec?lia Farias dos 11 June 2012 (has links)
Made available in DSpace on 2014-12-17T14:10:27Z (GMT). No. of bitstreams: 1 MariaCFS_DISSERT.pdf: 790525 bytes, checksum: c565e1057a72e1ddf23dec1d882788e8 (MD5) Previous issue date: 2012-06-11 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Toxoplasmosis is a zoonosis caused by Toxoplasma gondii, a protozoan that has a cosmopolitan geographic distribution and low host specificity. Usually a benign and selflimiting, infection can manifest itself in a severe systemic becoming overwhelming in fetuses and patients with immunosuppression. Domestic fowl are considered one of the most important hosts in the epidemiology of toxoplasmosis, since they are potential sources of infection for humans, in addition to playing the role of important indicators of environmental contamination by oocysts of T. gondii. We studied the prevalence of infection by the protozoan in chickens of different breeding systems mesoregions from the states of Rio Grande do Norte and Paraiba: broilers from commercial farms (200/PB) and free-range chickens of small farms (322/RN and PB). Were standardized IFAT and ELISA techniques for detecting specific antibodies in blood samples of birds, and commercial kit was used to determine the prevalence by IHAT. There was no seropositive reaction by T. gondii in the samples of broilers tested, indicating that the particularities of intensive management limit the chances of infection for these animals. Among the hens, the frequency of IgG anti-T. gondii diagnosed by the techniques of IHAT, IFAT and ELISA, respectively, were 3.73% (12/322), 37.88% (122/322) and 40.37% (130/322), for both young and adult animals. Amongst the seropositive samples by IFAT, 33 (27.05%) were positive at a dilution of 1:16, in 1:32, 31 (25.41%), in 1:64, 24 (19.67%), 15 (12.29%) in 1:128, and 19 presented titer greater than or equal to 1:256 (15.57%). The evaluation of the presence of anti-T. gondii should be careful, and reagents IHAT provided erratic results in this measure for the specie studied. This suggests the need for own standardization of the kit before the use in epidemiological studies in animal species. On the other hand, substantial agreement observed between IFAT and ELISA techniques (Kappa = 0.62) enables these methods as effective methodologies for the diagnosis of toxoplasmosis in chickens. The high prevalence of specific antibodies among poultry in the region studied attempts to the potential risk of transmission of toxoplasmosis to humans / A toxoplasmose ? uma zoonose causada pelo Toxoplasma gondii, protozo?rio que tem distribui??o geogr?fica cosmopolita e pouca especificidade parasit?ria. Comumente de curso benigno e autolimitante, a infec??o pode manifestar-se de forma sist?mica grave, tornando-se grav?ssima em fetos e pacientes com imunodepress?o. Galinhas dom?sticas s?o consideradas um dos mais importantes hospedeiros na epidemiologia da toxoplasmose, uma vez que s?o potenciais fontes de infec??o para humanos, al?m de desempenharem o papel de importantes indicadores da contamina??o ambiental por oocistos de T. gondii. Neste trabalho, estudou-se a frequ?ncia da infec??o pelo protozo?rio em galin?ceos de diferentes sistemas de cria??o provenientes de mesorregi?es dos estados do Rio Grande do Norte e Para?ba, tanto frangos de corte de granjas comerciais (200/PB), como galinhas caipira de pequenas propriedades rurais (322/RN e PB). Foram padronizadas t?cnicas de RIFI e ELISA para a detec??o de anticorpos s?ricos espec?ficos nas amostras sangu?neas das aves, e foi utilizado kit comercial para determina??o dessa preval?ncia pelo HAI. N?o foi observada infec??o por T. gondii em nenhuma das amostras de frango de corte analisada, indicando que particularidades do manejo intensivo limitam as chances de infec??o para esses animais. Entre as galinhas caipira, a frequ?ncia de anticorpos IgG anti-T. gondii diagnosticada pelas t?cnicas de HAI, RIFI e ELISA foi, respectivamente, 3,73% (12/322), 37,88% (122/322) e 40,37% (130/322), analisando animais jovens e adultos. Das amostras soropositivas pela RIFI, 33 (27,05%) foram reagentes na dilui??o 1:16; em 1:32, 31 (25,41%); em 1:64, 24 (19,67%); 15 (12,29%) em 1:128 e 19 apresentaram titula??o maior ou igual a 1:256 (15,57%). A avalia??o da presen?a de anticorpos anti-T. gondii deve ser criteriosa, sendo que os reagentes do HAI forneceram resultados err?ticos nesta medida, para a esp?cie estudada, sugerindo a necessidade de padroniza??o pr?pria dos kits para diagn?stico antes do uso em estudos epidemiol?gicos em esp?cies animais. Por outro lado, a concord?ncia substancial observada entre as t?cnicas RIFI e ELISA (Kappa = 0,62) capacita estas metodologias como t?cnicas eficazes no diagn?stico da infec??o pelo protozo?rio em galin?ceos. A alta frequ?ncia de anticorpos espec?ficos observada entre as aves da regi?o estudada atenta para o risco potencial de transmiss?o da toxoplasmose para o homem
198

Detecção imunoistoquímica de linfócitos T (CD3+) e B (CD79+) no encéfalo de cães com leishmaniose visceral e presença de anticorpos séricos anti-Toxoplasma gondii e anti-Neospora caninum /

Sakamoto, Keila Priscilla. January 2009 (has links)
Orientador: Gisele Fabrino Machado / Banca: Mary Marcondes / Banca: Rosemeri de Oliveira Vasconcelos / Resumo: A Leishmaniose visceral é uma enfermidade que possui uma grande variabilidade de manifestações clínicas, em humanos como em cães. Cães cronicamente infectados podem desenvolver desordens neurológicas, contudo, há poucos relatos que caracterizam as lesões e elucidam a patogenia da leishmaniose cerebral canina. Considerando a imunossupressão associada à leishmaniose visceral e que os patógenos oportunistas Toxoplasma gondii e Neospora caninum podem colaborar para a ocorrência de lesões no sistema nervoso central de cães naturalmente infectados por Leishmania chagasi, as populações de linfócitos B (CD79+) e T (CD3+) foram avaliadas no tecido nervoso de cães portadores de leishmaniose visceral e que possuem soropositividade para T. gondii e N. caninum. Lesões inflamatórias, caracterizadas por acúmulos de células mononucleares compostos principalmente por linfócitos T CD3+ predominaram em diversas regiões encefálicas dos cães infectados (P = 0,0012). Linfócitos B CD79+ foram detectados em pequena intensidade, não havendo diferença entre os grupos (P = 0,3604). Os resultados obtidos sugerem que a co-presença de leishmaniose visceral, toxoplasmose e neosporose é importante para o agravamento das lesões encefálicas, e que a imunossupressão gerada pela infecção por Leishmania não somente favorece a infecção por outros patógenos, mas colabora com esses, ocasionando lesões mais severas no tecido nervoso / Abstract: Visceral leishmaniasis is a disease with great variability regarding the clinical manifestations, in humans as in dogs. Chronically infected dogs may develop neurological disorders, however, there are few reports that characterise the lesions and make clear the pathogenesis of the canine cerebral leishmaniasis. Considering the immunossupression associated to visceral leishmaniasis and that the opportunist pathogens Toxoplasma gondii and Neospora caninum may collaborate to the occurrence of lesions in the central nervous system of dogs naturally infected by Leishmania chagasi, we evaluated the population of B (CD79+) and T (CD3+ ) lymphocytes in the nervous tissue of dogs with visceral leishmaniasis and with seropositivity to T. gondii and to N. caninum. Inflammatory lesions, characterised by mononuclear cellsaccumulation, composed mainly by CD3+ T lymphocytes predominated in several encephalic regions of the dogs from the infected groups (P=0.0012). CD79+ B lymphocytes were detected in very small intensity and presented no difference among groups (P=0.3604). The results presented herein suggest that the co-presence of visceral leishmaniasis, toxoplasmosis and neosporosis is important for the worsening of the encephalic lesions, and that the immunossupression caused by Leishmania infection not only propitiates the infection by other pathogens, but collaborate with them, causing more severe lesions in the brain / Mestre
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Detecção molecular de coccídios da familia sarcocystidae em amostras teciduais de pequenos felídeos neotropicais do Rio Grande do Sul / Molecular detection of coccidia family Sarcocystidae in tissues samples of small Neotropical wildlife felids of Rio Grande do Sul

Cañón-Franco, William Alberto January 2013 (has links)
Poucos estudos quantificam o risco relativo da saúde humana na transmissão spillover de doenças zoonóticas de populações de animais silvestres, estudos cruciais na compreensão da história natural das zoonoses. Coccídios, em particular os da família Sarcocystidae, são importantes agentes transmissíveis na interface homens - animais domésticos e silvestres. O diagnóstico da coccidiose é prejudicado pela limitada disponibilidade de amostras resultantes de populações animais de espécies em risco de extinção. O objetivo deste estudo foi detectar através da amplificação do locus ITS-1, protozoários das subfamílias Sarcocystinae e Toxoplasmatinae em amostras teciduais de Puma yagouaroundi, Leopardus geoffroyi, L. tigrinus, L. wiedii, L. colocolo e L. pardalis, depositados em coleções biológicas do Rio Grande do Sul, Brasil. Um objetivo adicional foi a obtenção de informações que permitissem avaliar o papel epidemiológico dos protozoários no ciclo silvestre dos parasitas e seu possível impacto sobre as populações de animais silvestres e na saúde pública. Noventa pequenos felídeos neotropicais de vida livre, representando seis espécies, foram amostrados. Destes, 31 felídeos (34,4%), de todas as seis espécies, foram positivos para Toxoplasma gondii e DNA foi detectado em 63 das 433 (14,6%) amostras de tecidos primários coletados a partir de língua (28,6%), cérebro (18,6%), músculo esquelético (17,1%), musculatura ocular (13,6%), globo ocular (13,6%), coração (11,1%), diafragma (5,4%) e humor vítreo (4,5%). Doze amostras primárias positivas ao T. gondii foram genotipadas com os marcadores moleculares SAG1, SAG2, SAG3, BTUB, GRA6, c22- 8, c29-2, L358, PK1, Apico e CS3 e a técnica multilocus PCR-RFLP, a amostra Py#36m foi totalmente caracterizada como do tipo I com alelo II no BTUB e um novo genótipo atípico Py#21M, ambos isolados de Puma yagouaroundi e nunca descrito no Brasil. Nove outras amostras tiveram caracterização parcial. Treze dos 90 felídeos foram positivos para Sarcocystis spp. (14,4%) e outros 18 felídeos, representando cinco espécies albergaram S. felis-like [Py (#75m, #83m, #35m, #20li, #55li), Lg (#80m, #70m, #88m, #71li, #67mOi), Lt (#19m, #48m, #89m, #84m), Lw (#12, #73d) e Lc (#82m, #76m)]. Um único felino de L. pardalis foi negativo. DNA do parasita foi detectado em 11,8% dos tecidos examinados (51/433): musculatura esquelética (26,5%), língua (23,2%), musculatura ocular (13,6%), diafragma (10,7%), cérebro (2,3%), coração (1,6%) e globo ocular (4,5%), nenhuma das 44 amostras de humor vítreo foi positiva. Esta é a primeira detecção e caracterização genética de T. gondii e de S. felislike em felídeos silvestres brasileiros de vida livre, demonstrando a presença destes agentes no ciclo silvestre e, a potencial transmissibilidade ao homem e a outros animais domésticos e silvestres. O uso de amostras de tecidos de animais silvestres depositados em coleções biológicas para estudos epidemiológicos de doenças monstraram serem de grande utilidade. / Few studies have quantified the relative risk of human health from spillover of zoonotic diseases from populations of wild animals; these studies are crucial for understanding the natural history of zoonoses. Coccidia, particularly from the family Sarcocystidae, are important transmissible agents at the interface of man and domestic and wild animals. The diagnosis of Coccidiosis is hampered by the limited availability of samples resulting from protection of natural populations of the species at risk of extinction. The aim of this study was to detected, by amplification of ITS-1 locus, protozoa from the subfamilies Sarcocystinae and Toxoplasmatinae in tissue samples from Puma yagouaroundi, Leopardus geoffroyi, L. tigrinus, L. wiedii, L. colocolo and L. pardalis, deposited in biological collections of the State of Rio Grande do Sul, Brazil. An additional aim was to obtain information that would enable assessment of the epidemiological role of the protozoa in the sylvatic cycle of the parasite, and its possible impact on wildlife populations and public health. Ninety free-living small wild felines, representing 6 species, were sampled. Of these, 31 felids (34.4%) of all six species were positive for T. gondii and DNA was detected in 63 of 433 (14.6%) primary tissue samples collected from the tongue (28.6%), brain (18.6%), skeletal muscle (17.1%), ocular muscles (13.6%), eye (13.6%), heart (11.1%), diaphragm (5.4%) and vitreous humor (4.5%). Twelve primary samples positive for T. gondii were genotyped with molecular markers SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and apical CS3. Using the multilocus PCR-RFLP technique, sample Py#36m was fully genotyped as Type I with allele II in locus BTUB, and a new atypical Py#21M, both isolates from Puma yagouaroundi and never described in Brazil. Nine other samples had a partial characterization. Thirteen of the 90 felids were positive for Sarcocystis spp. (14.4%) and another 18 felids, representing 5 species, harbored S. felis-like organisms [Py (#75m, #83m, #35m, #20li, #55li), Lg (#80m, #70m, #88m, #71li, #67mOi), Lt (#19m, #48m, #89m, #84m), Lw (#12, #73d) and Lc (#82m, #76m)]. A single felid of L. pardalis was negative. Parasite DNA was detected in 11.8% (51/433) of the tissues examined: muscle skeletal (26.5%), tongue (23.2%), ocular muscles (13.6%), diaphragm (10.7 %), brain (2.3%), heart (1.6%) and eye (4.5%); none of the 44 samples of vitreous humor was positive. This is the first description of the detection and genetic characterization of T. gondii and S. felis-like in free-living Brazilians wild felids, demonstrating the presence of these agents in the sylvatic cycle, and the potential transmition to humans and other domestic and wild animals. The use of tissue samples from wild animals deposited in biological collections for epidemiological studies of diseases demonstrated to be of great utility.
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Ocular toxoplasmosis : immunopathology and virulence : the influence of parasite virulence on the clinical, biological, and immunological characteristics of ocular toxoplasmosis (OT) in the Old and New World / La toxoplasmose oculaire : immunopathologie et Virulence : l'influence de la virulence du parasite sur les caractéristiques cliniques, biologiques et immunologiques de la toxoplasmose oculaire (OT) de l'Ancien et du Nouveau monde

De la Torre Cifuentes, Ligia Alejandra 18 September 2013 (has links)
Nous avons sélectionné des patients atteints d’une TO biologiquement confirmée et avons exploré les différences cliniques et biologiques de deux groupes de patients, l’un en France, l’autre en Colombie. Dans notre hypothèse de départ, les souches sud-américaines, seraient plus virulentes et elles pourraient jouer un rôle crucial dans la sévérité et l’évolution de la TO. Nous avons constaté, chez les patients colombiens, de plus grandes lésions de la rétine et une plus grande proportion de lésions maculaires, dans un contexte inflammatoire vitréen plus sévère. Le cytoquinome oculaire confirme une forte réponse inflammatoire chez les patients européens centrée sur l’IL-17, mais cette réponse Th17 est absente chez les sujets colombiens. L’IL-6 et l’IL-13 sont au contraire fortement augmentées chez ces derniers. Nous avons également démontré que certaines cytokines étaient associées à certaines caractéristiques cliniques comme la sévérité de l’inflammation ou la récurrence. Des travaux préliminaires nous ont permis de débuter une modélisation de ces affactions oculaires en employant une souche de type II et une souche atypique de T. gondii. Nous avons aussi évalué la possibilité d’effectuer des traitements ciblés en intraoculaires par transfection in vivo. Conclusion: Nous avons constaté des différences cliniques et biologiques entre les patients colombien et français. Il semble y avoir une régulation souche dépendante de la production d’IFN-y et d’IL-17. Ces différences pourraient contribuer à expliquer la plus grande sévérité des toxoplasmoses oculaires en Colombie. En se basant sur nos résultats nous pouvons envisager d’explorer des traitements immunomodulateurs plus ciblés. / Ocular involvement, mainly retinochoroiditis, is one of the most severe sequelae of Toxoplasma gondii infection. However, the pathophysiological mechanisms of retinal destruction are poorly understood. Several studies suggested a more frequent and more severe ocular involvement in South American infections compared with European infections, probably due to different T. gondii strains (Type I/III, and atypical vs. Type II). To compare the clinical characteristics and biological and immunological responses in a single study and using the same parameters, in Colombian and French patients with active ocular toxoplasmosis (OT), as well as to study the local cytokinome in aqueous humor of these patients and correlate it with the clinical features. We prospectively collected and compared the clinical features of patients with active OT, evaluated at the Department of Ophthalmology of Strasbourg University Hospital and of Quindio University Health-Center. Results of biological tests in the collected aqueous humor samples were compared between Colombian and French patients: the pattern of protein recognition by immunoblotting (IB); the relative diagnostic sensitivities of IB and Polymerase Chain Reaction (PCR); and the cytokine and chemokine profiles. We found that Colombian and French OT patients presented not only different clinical characteristics but also biological characteristics, and that more virulent South American strains might be responsible for these differences, due to a disruption of the protective effects of interferon gamma (IFN-γ). Retinal lesions were 50% greater in Colombian patients. Macular localization leading to visual impairment was observed in 56% of Colombian cases, compared with 13% of French patients. Moreover, more vitreous inflammation and vasculitis were observed in Colombian patients. However, cytokine assays of the aqueous humor showed upregulation of inflammatory responses in European patients, notably IL-17, which we did not observe in Colombian patients. In a mouse model, intraocular tachyzoite injection of type II and atypical T. gondii strains resulted in differences in parasite multiplication and pathology similar to those observed in human infections. Production of IL-17 and other inflammatory markers, like IL-6, MCP-1, and the Th17 transcription factor ROR-γt was observed upon infection with the type II PRU strain, but was much less with the atypical LEF strain. In a previous work, the cytokine and mRNA patterns showed an upregulation of Th1 responses, notably IFN-γ production, in French patients, and anti-IL-17A antibody markedly diminished clinical damage and retinal inflammation, and also diminished parasite proliferation. In contrast to these previous findings in French patients, the cytokinome of aqueous humor of OT Colombian patients showed a downregulation of Th1 and Th17 responses and an upregulation of the Th2 response. Correlation between the clinical characteristics of Colombian patients with active OT and the levels of cytokines in aqueous humor (AH) showed that local production of cytokines differed between patients with OT, and particular cytokine levels were related to more severe clinical characteristics. Some cytokines were related to a higher number of recurrences.There are clinical and biological differences between Colombian and French patients with OT. There seem to be strain-specific differences in IL-17 and IFN-γ induction, which play an important role in the pathogenesis of this disease. These differences should be considered when thinking in perspectives of any possible immune-modulatory treatment in OT.

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