Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2018-10-19T18:19:28Z No. of bitstreams: 1 2013 - Andresa Guimar?es.pdf: 1221110 bytes, checksum: 80ca8363650edbb042caea761cb85083 (MD5) / Made available in DSpace on 2018-10-19T18:19:28Z (GMT). No. of bitstreams: 1 2013 - Andresa Guimar?es.pdf: 1221110 bytes, checksum: 80ca8363650edbb042caea761cb85083 (MD5) Previous issue date: 2013-08-21 / CAPES / CNPq / Studies of domestic cats are little discussed in Brazil, especially regarding hemoparasita research where cats can be carriers of the agent and contribute to infection of man. Anaplasmataceae agents are gram-negative bacteria found in leukocytes and platelets, in which morulae characteristic of the infection is observed. The transmission of these agents to humans and animals occurs by arthropod vectors. Sorologic diagnostic methods are emphasized, but recent studies using molecular techniques demonstrate greater sensitivity and specificity, with better characterization of the agent. The present study aims to diagnose domestic cats in Rio de Janeiro naturally infected by agents Anaplasmataceae (Ehrlichia sp., and Anaplasma sp.) by serological and molecular methods, as well as evaluate hematological changes associated with these infections to guide veterinarians in the diagnosis of these diseases. Samples of 216 animals from clinics located in cities in the metropolitan region of Rio de Janeiro were used. The research of antibodies IgG anti-E. canis was performed by Immunofluorescence Assay (IFAT) and molecular detection of Ehrlichia sp. by nested Polymerase Chain Reaction (PCR) based on 16S rRNA gene. The results demostrate that 18 (8.3%) animals were positive in the direct detection by blood smear, 57 (26.4%) seropositive and 37 (17.1%) positive by nested-PCR for Ehrlichia sp., being 91 (42.1%) positive in at least one technique. The diagnosis of A. platys was performed by real-time PCR (qPCR) and nested-PCR based on 16S rRNA and gltA genes, respectively. Seventeen animals (7.9%) showed inclusions in platelet, eight (3.7%) were positive in qPCR, two (0,9%) in nested-PCR and 23 (10.6%) cats were considered positive for A. platys in at least one technique. The detection of A. phagocytophilum was performed by qPCR based on gene msp2, however, positive animals were not observed. The most common hematologic findings in animals positive for agents Anaplasmataceae were thrombocytopenia, leukocytosis, neutrophilia, regenerative left shift, anemia and hyperproteinemia. Only one animal showed co-positivity for both agents. Regarding the associated factor, only age interfered with positivity for Ehrlichia sp., in wich younger IFAT positive animals were less prone to infection, unlike that observed in the nested-PCR in which the younger had higher positivity. Only two positive samples in the nested-PCR for E. canis were subjected to sequencing. One sequence demonstrated 100% similarity with isolates of Ehrlichia sp. and E. canis and another showed 99% similarity with isolates of Ehrlichia sp. ocelot and Ehrlichia sp. little-spotted-cat. The two positive samples in the nested-PCR for A. platys demonstrated 100% similarity with isolates of A. platys reported in advanced. The present work demonstrates the circulation of agents Anaplasmataceae in domestic cats in Rio de Janeiro. / Estudos com felinos dom?sticos s?o pouco abordados no Brasil, principalmente em rela??o ? pesquisa de hemoparasitoses, em que os gatos podem ser portadores do agente e contribuir para infec??o do homem. Os agentes Anaplasmataceae s?o bact?rias gram-negativas que s?o encontradas em leuc?citos, plaquetas ou eritr?citos, formando m?rulas, caracter?sticas de sua infec??o. A transmiss?o desses agentes para o homem e animais ocorre principalmente por vetores artr?podes. Como m?todo diagn?stico ressalta-se a sorologia, por?m estudos recentes utilizando diagn?stico molecular demonstram maior sensibilidade e especificidade, com melhor caracteriza??o do agente. O presente estudo tem como objetivo diagnosticar felinos dom?sticos do Rio de Janeiro infectados naturalmente por agentes Anaplasmataceae (Ehrlichia sp. e Anaplasma sp.) por meio de m?todos sorol?gico e molecular, bem como avaliar as altera??es hematol?gicas associadas a estas infec??es. Foram utilizadas amostras de 216 animais, provenientes de cl?nicas localizadas em cidades da Regi?o Metropolitana do Rio de Janeiro. A pesquisa de anticorpos IgG anti-E. canis foi realizada pela Rea??o de Imunofluoresc?ncia Indireta (RIFI) e a detec??o molecular de Ehrlichia sp., pela nested Rea??o em Cadeia da Polimerase (nested-PCR) baseada no gene 16s rRNA. Os resultados demonstraram que 18 (8,3%) animais foram positivos na detec??o direta pelo esfrega?o sangu?neo, 57 (26,4%) soropositivos e 37 (17,1%) positivos na nested-PCR para Ehrlichia sp., sendo 91 (42,1%) considerados positivos para Ehrlichia sp. em pelo menos uma t?cnica. O diagn?stico de A. platys foi efetuado pela PCR em tempo real (qPCR) e nested-PCR baseada nos genes gltA e 16S rRNA, respectivamente. Dezessete animais (7,9%) apresentaram inclus?es em plaquetas, oito (3,7%) foram positivos na qPCR, dois (0,9%) na nested-PCR e 23 (10,6%) animais foram considerados positivos para A. platys em pelo menos uma t?cnica. A pesquisa de A. phagocytophilum foi realizada por meio da qPCR baseada no gene msp2; no entanto, n?o foram observados animais positivos. Os achados hematol?gicos mais frequentes nos animais positivos para agentes Anaplasmataceae foram trombocitopenia, leucocitose, neutrofilia, desvio ? esquerda regenerativo, anemia e hiperproteinemia. Apenas um animal apresentou co-positividade para ambos os agentes. Em rela??o aos dados associados, somente a idade interferiu na positividade para Ehrlichia sp., sendo na RIFI os animais mais jovens menos predispostos ? infec??o, ao contr?rio do observado na nested-PCR em que os mais jovens apresentaram maior ?ndice de positividade. Apenas duas amostras positivas na nested-PCR para E. canis foram submetidas ao sequenciamento, sendo que uma sequ?ncia apresentou 100% de similaridade com isolados de Ehrlichia sp. e E. canis e a outra demonstrou 99% de similaridade com isolados de Ehrlichia sp. ocelot e Ehrlichia sp. little-spotted-cat. As duas amostras positivas na nested-PCR para A. platys apresentaram 100% de similaridade com isolados de A. platys previamente descritos. O presente trabalho demonstra, portanto, a ocorr?ncia de agentes Anaplasmataceae na popula??o de felinos dom?sticos do Rio de Janeiro.

Made available in DSpace on 2016-04-28T20:18:30Z (GMT). No. of bitstreams: 1 2008 - Erica Mateus Toledo Accetta.pdf: 809163 bytes, checksum: 04d79d0b9854d1e99fd77cff6ae68a4f (MD5) Previous issue date: 2009-08-27 / Canine ehrlichiosis is an important infectious disease whose prevalence has been increasead in most areas of Brazil. Clinical signals and the laboratorial findings are variable. The present work had as objective to determine the frequency of the infection for E. canis and Anaplasma platys in dogs with trombocytopenia at Regi?o dos Lagos - State of Rio de Janeiro. It has been evaluated the CBC of 1127 dogs with trombocytopenia, a total of 3019 laboratorial tests carried through in the period of June 2006 and July 2007, at CEVET Lagos Lab, providing atendence to several clinics of Araruama, Iguaba Grande, S?o Pedro d Aldeia, Cabo Frio, Arraial do Cabo and B?zios. Erliquiosis was diagnosised through haemoparasit in smears of total blood s research stained with Panoptic kit. Eighty-four dogs (7.45%) were considered infected by the discovery of morulae of Ehrlichia spp. e Anaplasma platys. Normocytic normochromic anaemia and monocytosis were the most hematological alterations found. / A erliquiose canina ? uma importante doen?a infecciosa cuja preval?ncia tem aumentado significativamente em v?rias regi?es do Brasil. Os sinais cl?nicos e os achados laboratoriais s?o vari?veis. O presente trabalho teve como objetivo determinar a freq??ncia da infec??o por E. canis e A. platys em c?es com trombocitopenia na Regi?o dos Lagos do Estado do Rio de Janeiro. Foram avaliados os exames hematol?gicos de 1127 c?es com trombocitopenia de um total de 3019 exames realizados no per?odo de junho de 2006 a julho de 2007, no laborat?rio CEVET Lagos, que presta atendimento ?s diversas cl?nicas dos munic?pios de Araruama, Iguaba Grande, S?o Pedro d Aldeia, Cabo Frio, Arraial do Cabo e B?zios. A erliquiose foi diagnosticada atrav?s da pesquisa de hemoparasitos em esfrega?os de sangue total corados com kit pan?ptico. O diagn?stico baseou-se no achado de m?rulas de Ehrlichia spp. e Anaplasma platys, sendo considerados infectados 84 (7,45%) c?es. Anemia normoc?tica normocr?mica e monocitose foram as altera??es hematol?gicas mais freq?entes.

Made available in DSpace on 2016-04-28T20:16:32Z (GMT). No. of bitstreams: 1 2006-Valeria Regia Franco Sousa.pdf: 1545960 bytes, checksum: 07990d273ccfe1ac6c38929e5d3d100c (MD5) Previous issue date: 2006-02-20 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / This work had for objective to investigate the infection for Ehrlichia canis and Anaplasma platys in the dogs taken care of in the Veterinarian Hospital of the Universidade Federal de Mato Grosso, through the examination of blood smears and of the PCR, analyzing the clinical and laboratories findings. During the period of May of 2004 the July of 2005, 195 dogs with suggestive clinical signs of ehrlichiosis or had contact with the infection had been examined. The canine cyclic thrombocytopenia, caused for the A. platys, provokes reduction of platelets to each 14 days, without provoking severe signs, but when associated to the infection for E. canis the clinical gravity it increases. In the thirteen dogs taken care of with positive PCR for E. canis diagnosised by the PCR it was possible to verify the diversity of signals, with statistical significant predominance of the hemorrhagic riots. Already in the 195 dogs tested for the examination of blood smears it did not have predominance of no clinical signs. The hematologic findings had also been nonspecific, occurring in such a way anemia, leukopenia and thrombocytopenia, how much normality or increase of the cells. The alterations observed in the analysis biochemist had not been exclusive of the groups with infection. In these groups increase of plasma proteins occurred, with hyperglobulinaemia, without, however to have significant difference, despite this finding being frequent in ehrlichiosis. From the analysis of the PCR it was confirmed that the infection for E. canis and A. platys occur in the dogs taken care of in the Veterinarian Hospital of the UFMT, having to be taken writ of prevention, mainly in the control of the vector, since all the ticks found in the examined dogs had been of species Rhipicephalus sanguineus. / Este trabalho teve por objetivo investigar a infec??o por Ehrlichia canis e Anaplasma platys nos c?es atendidos no Hospital Veterin?rio da Universidade Federal de Mato Grosso, atrav?s das t?cnicas de esfrega?o sang??neo e de PCR, analisando os achados cl?nicos e laboratoriais. Durante o per?odo de maio de 2004 a julho de 2005 foram examinados 195 c?es com sinais cl?nicos sugestivos de erlichiose ou com contactantes com a infec??o. A trombocitopenia canina c?clica, causada pelo A. platys, provoca diminui??o das plaquetas a cada 14 dias, sem provocar sinais severos, mas quando associada ? infec??o por E. canis a gravidade cl?nica aumenta. Nos treze c?es atendidos com PCR positiva para E. canis foi poss?vel verificar a diversidade de sinais, com predomin?ncia estatisticamente significativa dos dist?rbios hemorr?gicos. J? nos 195 c?es testados pela t?cnica de esfrega?o sang??neo n?o houve predom?nio de nenhum sinal cl?nico. Os achados hematol?gicos tamb?m foram inespec?ficos, ocorrendo tanto anemia, leucopenia e trombocitopenia, quanto normalidade ou aumento das c?lulas. As altera??es observadas na an?lise bioqu?mica n?o foram exclusivas dos grupos com infec??o. Nestes grupos ocorreu aumento das prote?nas plasm?ticas, com hiperglobulinemia, sem, no entanto haver diferen?a significativa, apesar deste achado ser freq?ente na erliquiose. A partir da an?lise da PCR confirmou-se que a infec??o por E. canis e A. platys ocorre nos c?es atendidos no Hospital Veterin?rio da UFMT, devendo ser tomadas medidas preventivas, principalmente no controle do vetor, j? que todos os carrapatos encontrados nos c?es examinados foram da esp?cie Rhipicephalus sanguineus.

Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2017-10-19T13:49:16Z No. of bitstreams: 1 2016 - Claudia Bezerra da Silva.pdf: 8032175 bytes, checksum: ef71dd2a0e7801e9000e116c822a3a00 (MD5) / Made available in DSpace on 2017-10-19T13:49:16Z (GMT). No. of bitstreams: 1 2016 - Claudia Bezerra da Silva.pdf: 8032175 bytes, checksum: ef71dd2a0e7801e9000e116c822a3a00 (MD5) Previous issue date: 2016-03-11 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / and investigate the circulation of this agent in dogs in the Itaguai microregion, Rio de Janeiro, Brazil, and dogs and ticks in two provinces of the island of Cuba, analyzing epidemiological aspects associated with infections caused by this bacterium in dogs. A new real-time polymerase chain reaction method (qPCR) was patterned to target the citrate synthase gene (gltA) for the identification of A. platys in naturally infected dogs. The primers and probe were designed to amplify a fragment of 84 base pairs based on gltA gene sequences of A. platys available in GenBank. 186 blood samples of dogs from Itaguai microregion, Rio de Janeiro, Brazil, were tested by qPCR. The same samples were tested by cytology and nested polymerase chain reaction (nPCR, 16S rDNA) to determine the performance of qPCR front of these techniques. 17.20% of the samples tested positive by qPCR were significantly more than that detected by nPCR (13.98%). The qPCR technique was more specific than cytology, due to false-positive results obtained by optical microscopy. The prevalence of A. platys in dogs from Itaguai microregion was 14.4%. Dogs less than six months, infested by ticks, that spend the most of the time restrict to domestic environment and without shelter are factors associated with infection by this hemoparasite in dogs in the study area. During research, A. platys held in Cuba, 100 blood samples were collected from residents dogs in four cities located in the provinces of Havana and Mayabeque. When inspecting the animals, found ticks were collected, identified and carefully grouped, forming a total of 49 pools. DNA extracted from blood samples from dogs and ticks were subjected nPCR (16S rDNA). Positive samples in nPCR were also subjected to conventional PCR (gltA gene), and the products were sequenced. Only the species Rhipicephalus sanguineus sensu lato was found in Cuban dogs and 10.2% (n=5/49) of these ticks added to 16.0% (n=16/100) dogs were considered positive for A. platys. All sequences analyzed of the gltA and 16S rDNA genes, respectively, showed a 99-100% identity with sequences from A. platys reported in other countries. Phylogenetic analysis showed two clusters defined for the 16S rDNA gene and three clusters defined for the gltA gene. Based on the gltA gene, the deduced amino acid sequence showed two points of non-synonymous mutations at positions 88 and 168 compared to the reference sequence DQ525687. A preliminary study on the epidemiological aspects associated with infection with A. platys showed no statistical association with the variables studied (p> 0.05). This study also to report the first evidence of A. platys in both dogs and ticks in Cuba also presents for the first time the development of a new qPCR method that contributes to the advancement of research involving A. platys. The epidemiological study in Brazil allowed us to identify significant factors in the occurrence of canine anaplasmosis, while in Cuba, it can be concluded that more research is needed to assess what the deciding factors in the transmission and spread of A. platys in that country. / platys, e investigar a circula??o deste agente em c?es na microrregi?o de Itagua?, Rio de Janeiro, Brasil, e c?es e carrapatos em duas prov?ncias da ilha de Cuba, analisando aspectos epidemiol?gicos associados ? infec??o causada por esta bact?ria em c?es. Um novo m?todo de rea??o em cadeia da polimerase em tempo real (qPCR) foi padronizado com alvo no gene citrato sintase (gltA) para a identifica??o de A. platys em c?es naturalmente infectados. Os oligoiniciadores e a sonda foram desenhados para amplificar um fragmento de 84 pares de base baseado em sequ?ncias do gene gltA de A. platys dispon?veis no GenBank. 186 amostras de sangue de c?es da microrregi?o de Itagua?, Rio de Janeiro, Brasil, foram testados pela qPCR. As mesmas amostras foram testadas pela citologia e rea??o em cadeia da polimerase nested (nPCR, 16S rDNA) para determinar o desempenho da qPCR frente ? essas t?cnicas. 17,20% das amostras testadas pela qPCR foram positivas, significativamente mais do que detectado pela nPCR (13,98%). A t?cnica de qPCR foi mais espec?fica que a citologia, em virtude dos resultados falsopositivos obtidos pela microscopia ?ptica. A preval?ncia de A. platys em c?es da microrregi?o de Itagua? foi de 14,4%. C?es com menos de seis meses, infestados por carrapatos, que possam maior tempo restrito ao ambiente dom?stico e sem abrigo s?o fatores associados a infec??o por este hemoparasito em c?es na regi?o do estudo. Durante investiga??o de A. platys realizada em Cuba, 100 amostras de sangue foram coletadas de c?es residentes em quatro cidades localizadas nas prov?ncias de Habana e Mayabeque. Ao inspecionar os animais, carrapatos encontrados foram coletados, identificados e criteriosamente agrupados, formando um total de 49 pools. Amostras de DNA extra?das do sangue dos c?es e de carrapatos foram submetidas a nPCR (16S rDNA). Amostras positivas na nPCR foram tamb?m submetidas a PCR convencional (gene gltA), e os produtos foram sequenciados. Somente a esp?cie Rhipicephalus sanguineus sensu lato foi encontrada em c?es cubanos, e 10,2% (n=5/49) desses carrapatos somado aos 16,0% (n=16/100) de c?es foram considerados positivos para A. platys. Todas as sequ?ncias analisadas dos genes gltA e 16S rDNA, respectivamente, mostraram uma identidade de 99-100% com sequ?ncias de A. platys reportadas em outros pa?ses. A an?lise filogen?tica mostrou dois clusters definidos para o gene 16S rDNA e tr?s clusters definidos para o gene gltA. Com base no gene gltA, a sequ?ncia de amino?cidos deduzidos demonstrou dois pontos de muta??es n?o-sin?nimas nas posi??es 88 e 168 comparados com sequ?ncia de refer?ncia DQ525687. Um estudo preliminar sobre os aspectos epidemiol?gicos associados com a infec??o por A. platys demonstrou nenhuma associa??o estat?stica com as vari?veis avaliadas (p > 0,05). O presente estudo al?m de relatar a primeira evid?ncia de A. platys em ambos c?es e carrapatos em Cuba, tamb?m apresenta pela primeira vez o desenvolvimento de um novo m?todo de qPCR que contribui para o avan?o da pesquisa envolvendo A. platys. O estudo epidemiol?gico realizado no Brasil permitiu identificar fatores importantes na ocorr?ncia da anaplasmose canina, enquanto em Cuba, pode-se concluir que mais investiga??es s?o necess?rias para avaliar quais os fatores decisivos na transmiss?o e dispers?o de A. platys nesse pa?s.