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dc.contributor.authorSantos, Huarrisson Azevedo
dc.date.accessioned2023-12-21T18:43:26Z-
dc.date.available2023-12-21T18:43:26Z-
dc.date.issued2011-04-11
dc.identifier.citationSANTOS, Huarrisson Azevedo. Diagnóstico molecular de Anaplasma phagocytophilum (Foggie, 1949) em cães domésticos e carrapatos de áreas urbanas e rurais na microrregião de Itaguaí, Estado do Rio de Janeiro. 2011. 76 f. Tese (Doutorado em Ciências Veterinárias) - Instituto de Veterinária, Universidade Federal Rural do Rio de Janeiro, Seropédica - RJ, 2011.por
dc.identifier.urihttps://rima.ufrrj.br/jspui/handle/20.500.14407/9744-
dc.description.abstractAnaplasma phagocytophilum é um patógeno emergente capaz de infectar humanos, equinos e cães em todo o mundo. Essa bactéria é transmitida por carrapatos ixodídeos e mantida na natureza por uma variedade de pequenos mamíferos silvestres. O objetivo do presente estudo foi avaliar a presença do DNA de A. phagocytophilum em cães e carrapatos da microrregião de Itaguaí, Rio de Janeiro, além de estabelecer os fatores associados (idade, sexo, raça, infestação por carrapatos, etc) e as possíveis alterações hematológicas com a infecção por A. phagocytophilum em cães desta região. Essa bactéria foi detectada pela primeira vez no Brasil, em cães e carrapatos de áreas rurais e urbanas da microrregião de Itaguai, RJ através da Reação em Cadeia da Polimerase (PCR) em tempo real. Dos 398 cães amostrados, 24 (6,03%) foram considerados positivos, com limite de detecção (CT) variando entre 26 a 35 ciclos. As amostras positivas clonadas em pGEM-T apresentaram 100% de identidade com outras sequências de A. phagocytophilum depositadas no GenBank. Utilizando números definidos de cópias do plasmídio contendo um inserto de 122pb do gene msp2 a sensibilidade analítica do ensaio de PCR em tempo real através do sistema SYBR green foi capaz de detectar até três cópias do plasmídio. O ensaio foi considerado específico quando DNA de bactérias proximamente relacionadas à A. phagocytophilum foi colocado na reação. Um fragmento de 122pb do gene msp2 de A. phagocytophilum foi detectado através da TaqMan- PCR em um carrapato fêmea da espécie Amblyomma cajennense e em cinco Rhipicephalus sanguineus, sendo quatro machos e uma fêmea. As amostras de carrapatos positivas obtiveram CT que variaram entre 26,75 e 33,14 ciclos. A técnica de PCR em tempo real utilizando o sistema TaqMan foi capaz de detectar uma única cópia do gene msp2 de A. phagocytophilum em amostras de DNA de carrapatos. Infestação por carrapatos foram observadas em 66,67% (n=16) dos cães positivos na PCR. As espécies de carrapatos identificadas nos cães positivos foram R. sanguineus e A. cajennense, com predominância desta última espécie de carrapato. O histórico de infestação por carrapatos, acesso a áreas de mata e condição de limpeza do ambiente foram os fatores que apresentaram associação com a presença do DNA de A. phagocytophilum em cães na região estudada. Os principais achados hematológicos em cães infectados com A. phagocytophilum foram trombocitopenia, linfopenia e aumento do volume plaquetário médio. Os resultados do presente estudo demonstram que o agente da Anaplasmose Granulocítica Canina circula na região estudada e os cães podem servir como fonte de infecção para carrapatos vetores do agente. Este é o primeiro relato de A. phagocytophilum no Brasil, um agente zoonótico emergente responsável pela Anaplasmose Granulocítica Humana.por
dc.description.sponsorshipCoordenação de Aperfeiçoamento de Pessoal de Nível Superior, CAPES, Brasil.por
dc.formatapplication/pdf*
dc.languageporpor
dc.publisherUniversidade Federal Rural do Rio de Janeiropor
dc.rightsAcesso Abertopor
dc.subjectAnaplasma phagocytophilumpor
dc.subjectCãespor
dc.subjectPCR em Tempo Realpor
dc.subjectHematologiapor
dc.subjectEpidemiologiapor
dc.subjectAnaplasma phagocytophilumeng
dc.subjectdogseng
dc.subjectreal-time PCReng
dc.subjectHematologyeng
dc.subjectEpidemiologyeng
dc.titleDiagnóstico molecular de Anaplasma phagocytophilum (Foggie, 1949) em cães domésticos e carrapatos de áreas urbanas e rurais na microrregião de Itaguaí, Estado do Rio de Janeiropor
dc.title.alternativeMolecular diagnostic of Anaplasma phagocytophilum (Foggie, 1949) in domestic dogs and ticks of urban and rural areas from Itaguai microregion, Rio de Janeiro stateeng
dc.typeTesepor
dc.description.abstractOtherAnaplasma phagocytophilum is an emerging pathogen of humans, horses, and dogs worldwide that is transmitted by Ixodid ticks and maintained in a variety of small wild mammal species.The objective of the present study was to evaluate the presence of A. phagocytophilum DNA in dogs and ticks of Itaguai microregion, Rio de Janeiro, as well as establish the associated factors (age, gender, breed, tick infestation, etc) and possible hematological alterations with A. phagocytophilum infection in dogs. Anaplasma phagocytophilum was detected for the first time from Brazil, in dogs the ticks of urban the rural areas in the Itaguai microregion, Rio de Janeiro state, by real-time polymerase chain reaction (qPCR). Of 398 samples, 24 (6.03%) were positive, with a CT ranging from 26 to 35 cycles. Amplicons from one positive sample were cloned in the pGEM-T and sequenced. The sequence obtained demonstrated 100% identity with other A. phagocytophilum sequences published in the GenBank database. The analytical sensitivity of RT-PCR using SYBR green system was able to detect three plasmid copies when defined numbers of plasmid copies containing 122 base pairs from the msp2 gene were used. The assay was considered specific when DNA from bacteria closely related to A. phagocytophilum was placed in the reaction. A fragment of 122pb msp2 gene of A. phagocytophilum was detected by TaqMan-PCR in one female tick of Amblyomma cajennense and in five Rhipicephalus sanguineus, being four males and one female. Positive tick samples obtained CT that ranged from 26.75 to 33.14 cycles. The TaqMan-PCR assay was capable to detect a single copy of A. phagocytophilum msp2 gene in DNA tick sample. Tick infestation was observed in 66.67% (n = 16) of PCR positive dogs. The tick species identified in positive dogs were R. sanguineus and A. cajennense, with predominance of the latter tick species. The history of tick infestation, access to forest areas and environment condition of cleanliness were factors associated with A. phagocytophilum infection in dogs. The main hematologic findings in dogs infected with A. phagocytophilum were thrombocytopenia, lymphopenia and increased mean platelet volume.These results demonstrate that the canine granulocytic anaplasmosis agent is present in regions in which dogs could be a source of infection for tick vectors. This is the first report of A. phagocytophilum in Brazil, a emergent zoonotic agent responsible of Human Granulocitic Anaplasmosis.eng
dc.contributor.advisor1Massard, Carlos Luiz
dc.contributor.advisor1ID257781297-34por
dc.contributor.advisor1Latteshttp://lattes.cnpq.br/7743112049924654por
dc.contributor.advisor-co1Baldani, Cristiane Divan
dc.contributor.advisor-co2Thomé, Sandra Maria Gomes
dc.contributor.referee1Leite, Romario Cerqueira
dc.contributor.referee2Salcedo, Joaquim Hernan Patarroyo
dc.contributor.referee3Machado, Rosangela Zacarias
dc.contributor.referee4Almosny, Nadia Regina Pereira
dc.creator.ID083833295-04por
dc.creator.Latteshttp://lattes.cnpq.br/3751609492049306por
dc.publisher.countryBrasilpor
dc.publisher.departmentInstituto de Veterináriapor
dc.publisher.initialsUFRRJpor
dc.publisher.programPrograma de Pós-Graduação em Ciências Veterináriaspor
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