Ocorrência de Ehrlichia e Anaplasma em roedores no Brasil
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Data
2017-02-14
Autores
Benevenute, Jyan Lucas [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
Embora novos genótipos de agentes Anaplasmataceae vêm sendo detectados em carnívoros selvagens, aves selvagens e cervídeos no Brasil, poucos são os estudos acerca da circulação destes patógenos em pequenos mamíferos em nosso território. O presente trabalho teve como objetivo investigar a presença de DNA de Ehrlichia e Anaplasma em roedores amostrados no Brasil. Entre 2000 e 2011, diferentes espécies de roedores [n = 60] foram capturadas em cinco biomas brasileiros. As amostras de baço de roedores foram submetidas a extração de DNA utilizando um kit comercial. A presença de DNA de Ehrlichia e Anaplasma foi investigada utilizando ensaios de PCR em tempo real e convencionais dirigidos a vários genes. Dentre as 458 amostras de baço de roedores testadas, 0,44% (2/458) mostraram-se positivas para Ehrlichia spp. e 2,40% (11/458) para Anaplasma spp., pelos ensaios de PCR em Tempo Real multiplex baseados no gene groESL. A quantificação média do número de cópias de um fragmento do gene groESL por microlitro variou de 1,071 x 102 a 2,808 x 102 cópias para Ehrlichia spp. e 1,123 x 100 a 1,310 x 102 para Anaplasma spp. Pelos ensaios de PCR convencional baseados no gene 16S rRNA, 1,09% (5/458) das amostras mostraram-se positivas para Ehrlichia sp. e 1,97% (9/458) para Anaplasma sp. Análises filogenéticas de máxima verossimilhança baseada em um fragmento de 546 pb do gene 16S rRNA posicionaram os genótipos de Anaplasma detectados em roedores próximos a A. phagocytophilum ou isolados de A. marginale e A. odocoilei. Por outro lado, os genótipos de Ehrlichia detectados em roedores amostrados mostraram-se intimamente relacionados com E. canis com base na análises filogenéticas de um fragmento de 358 pb do gene 16S rRNA. O presente estudo mostrou baixa ocorrência de Anaplasma e Ehrlichia em roedores no Brasil.
Although new genotypes of Anaplasmataceae agents have been detected in wild carnivores, wild birds and deer in Brazil, few reports have been done in order to assess the circulation of these pathogens in small mammals in our territory. The present work aimed to investigate the presence of Ehrlichia and Anaplasma in rodents sampled in Brazil. Between 2000 and 2011, different rodent species [n=60] were trapped in five Brazilian biomes. Rodents' spleen samples were submitted to DNA extraction using a commercial kit. The presence of Ehrlichia and Anaplasma DNA was investigated using real time and conventional PCR assays targeting several genes. Among 458 rodent tested spleen samples, 0.44% (2/458) were positive for Ehrlichia spp. and 2.40% (11/458) for Anaplasma spp., by a multiplex qPCR assay based on groESL gene. The mean quantification of the number of copies of 83pb groESL gene fragment per microliter ranged from 1,071 x 102 to 2,808 x 102 copies for Ehrlichia spp., and 1,123 x 100 to 1,310 x 102 for Anaplasma spp. Out of 458 samples tested, 1.09% (5/458) were positive for Ehrlichia sp. and 1.97% (9/458) for Anaplasma sp. by cPCR assays based on 16S rRNA gene. Maximum Likelihood phylogenetic analyse based on 546pb fragment of 16S rRNA gene positioned the Anaplasma genotypes detected in rodents near to A. phagocytophilum or A. marginale and A. odocoilei isolates. On the other hand, the Ehrlichia genotypes detected in sampled rodents were closely related to E. canis, according to the phylogenetic analyses based on 358pb 16S rRNA gene fragment. The present study showed a low occurrence of Anaplasma and Ehrlichia in rodents in Brazil.
Although new genotypes of Anaplasmataceae agents have been detected in wild carnivores, wild birds and deer in Brazil, few reports have been done in order to assess the circulation of these pathogens in small mammals in our territory. The present work aimed to investigate the presence of Ehrlichia and Anaplasma in rodents sampled in Brazil. Between 2000 and 2011, different rodent species [n=60] were trapped in five Brazilian biomes. Rodents' spleen samples were submitted to DNA extraction using a commercial kit. The presence of Ehrlichia and Anaplasma DNA was investigated using real time and conventional PCR assays targeting several genes. Among 458 rodent tested spleen samples, 0.44% (2/458) were positive for Ehrlichia spp. and 2.40% (11/458) for Anaplasma spp., by a multiplex qPCR assay based on groESL gene. The mean quantification of the number of copies of 83pb groESL gene fragment per microliter ranged from 1,071 x 102 to 2,808 x 102 copies for Ehrlichia spp., and 1,123 x 100 to 1,310 x 102 for Anaplasma spp. Out of 458 samples tested, 1.09% (5/458) were positive for Ehrlichia sp. and 1.97% (9/458) for Anaplasma sp. by cPCR assays based on 16S rRNA gene. Maximum Likelihood phylogenetic analyse based on 546pb fragment of 16S rRNA gene positioned the Anaplasma genotypes detected in rodents near to A. phagocytophilum or A. marginale and A. odocoilei isolates. On the other hand, the Ehrlichia genotypes detected in sampled rodents were closely related to E. canis, according to the phylogenetic analyses based on 358pb 16S rRNA gene fragment. The present study showed a low occurrence of Anaplasma and Ehrlichia in rodents in Brazil.
Descrição
Palavras-chave
Análise filogenética, Anaplasmataceae, Brasil, Caracterização molecular, PCR em tempo real quantitativa, Roedores