Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/311635
Type: TESE
Title: Detecção microbiológica e molecular da bacteremia por Bartonella spp. em gatos
Title Alternative: Molecular and microbiological detection of Bartonella spp. bacteremia in cats
Author: Drummond, Marina Rovani, 1985-
Advisor: Velho, Paulo Eduardo Neves Ferreira, 1966-
Abstract: Resumo: Atualmente o genero Bartonella compreende pelo menos 31 especies e subespecies, sendo 15 delas conhecidamente patogenicas ao homem. Tres especies de Bartonella estao associadas ao maior numero de manifestacoes clinicas em seres humanos. Sao elas: Bartonella bacilliformis, Bartonella quintana e Bartonella henselae. A B. henselae e a bacteria mais associada a doencas humanas e sua transmissao e muitas vezes relacionada ao trauma cutaneo causado pelo arranhao de gatos infectados. Em recente estudo, documentou-se que mais de dois por cento dos doadores de sangue da regiao de Campinas testados estavam bacteremicos por Bartonella spp. e o contato com animais foi um fator de risco para a aquisicao da infeccao. Com o objetivo de avaliar a prevalencia de bacteremia por Bartonella spp. e isolar uma cepa regional de B. henselae, e foram analisadas 112 amostras de sangue de gatos, sendo que destes, 84 (75%) eram nao domiciliados. A partir do sangue total coletado durante o procedimento cirurgico para castracao, foi realizada extracao de DNA, seguida de PCR nested que amplifica a regiao FtsZ e e especifica para B. henselae. Este sangue tambem foi inoculado em meio liquido BAPGM (Bartonella Alpha-Proteobacteria Growth liquid Medium). Apos dez dias de incubacao, parte desta cultura liquida de enriquecimento foi semeada em meio solido enriquecido com 30% de sangue de carneiro e parte analisada pela mesma PCR nested e por PCR simples especifica para o genero Bartonella e que amplifica a regiao ITS. As culturas solidas foram incubadas por ate 45 dias e as que apresentaram crescimento foram encaminhadas as duas diferentes reacoes de PCR ja descritas. O DNA de B. henselae foi detectado em 86 (77%) das 112 amostras de sangue e em 56 (50%) das amostras da cultura de liquida de enriquecimento. No total, a bacteremia foi detectada em 90% (101/112) dos gatos deste estudo. Constatou-se maior prevalencia entre gatos nao domiciliados (95%, 80/84) quando comparada com a dos gatos de proprietarios (75%, 21/28). A deteccao da bacteremia por meio de PCR simples, especifica para o genero Bartonella, foi possivel em 31 das 112 (28%) culturas liquidas de enriquecimento. Dezesseis isolados foram obtidos da cultura solida, sendo que 11 foram PCR positivos. As amostras foram sequenciadas e tres destas colonias, que demonstraram 100% de homologia com a cepa de B. henselae Brazil- 1 na regiao ITS analisada, foram depositadas na Colecao de Culturas do Instituto Adolfo Lutz, sendo as primeiras do Brasil. A PCR nested especie especifica foi positiva em, pelo menos, uma amostra de todos os gatos em que a bacteremia foi detectada. Estes resultados mostram que a bacteremia causada por Bartonella spp. e muito frequente nos gatos de Campinas e sugerem que a prevalencia da infeccao por Bartonella spp. nos gatos e suas consequencias para a saude publica tem sido subestimadas. Metodos diagnosticos mais sensiveis precisam ser utilizados na investigação desta infecção

Abstract: Currently, Bartonella genus comprises at least 31 species and subspecies, 15 pathogenic to humans. Three species are associated with the largest number of clinical symptoms in human beings: Bartonella bacilliformis, Bartonella quintana and Bartonella henselae. B. henselae is the one most frequently associated with human diseases. This specie is considered zoonotic and its transmission is usually related with infected cat scratches. In a recent study, bacteremia was detected in more than two percent of blood donors from Campinas area and contact with animals was documented as a risk factor for Bartonella spp. infection. In order to evaluate Bartonella spp. bacteremia prevalence in cats from Campinas and to isolate a B. henselae regional strain, we analyzed blood samples from 112 cats (75% stray cats). The whole blood that was collected during spay surgery was submitted to DNA extraction and tested with specie-specific FtsZ nested PCR for B. henselae. A pre-enrichment culture in liquid medium BAPGM (Bartonella Alpha- Proteobacteria Growth Liquid Medium) was also performed. After ten-day culture, an aliquot was seeded and incubated up to 45 days in a solid medium supplemented with 30% sheep blood and another aliquot was tested for two PCRs: specie-specific FtsZ nested PCR for B. henselae and Bartonella genus-specific ITS single tube PCR. Sample isolates obtained from solid cultures were also tested by the two different PCR reactions described above. B. henselae DNA was detected in 86 (77%) of 112 blood samples and 56 (50%) of pre-enrichment culture samples. In total, bacteremia was detected in 90% (101/112) cats. Higher bacteremia prevalence was detected in stray cats (95%, 80/84 cats) when compared to client-owned subjects (75%, 21/28 cats). When the genus-specific ITS single tube PCR was used, Bartonella spp. bacteremia was detected just in 31/112 (28%) of preenrichment culture. Sixteen Gram-negative isolates were obtained from solid medium culture and eleven of them were PCR positive. Some samples were sequenced and three of these isolates demonstrated a 100% homology with B. henselae Brazil-1 strain at analyzed ITS region. These isolates were the first samples of this strain to be deposited in Brazil, at Adolfo Lutz Institute Culture Collection. The specie-specific FtsZ nested PCR was positive at least at one sample of bacteremic cats. Our results show that Bartonella sp. bacteremia prevalence among cats is very frequent in Campinas and suggest that the prevalence of Bartonella spp. infection among cats and its consequences for public health remains underestimated. More sensitive diagnostic methods must be used in the study of this infection
Subject: Bartonella
Diagnóstico
Gatos
Reação em cadeia da polimerase
Language: Português
Editor: [s.n.]
Date Issue: 2012
Appears in Collections:FCM - Tese e Dissertação

Files in This Item:
File SizeFormat 
Drummond_MarinaRovani_M.pdf532 kBAdobe PDFView/Open


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.