Molecular detection of Anaplasmataceae agents in Dasyprocta azarae in northeastern Brazil.

Recently, the importance of wild-living rodents for maintenance of pathogens of the family Anaplasmataceae in the environment was investigated. These mammals play a role as reservoirs for these pathogens and act as hosts for the immature stages of tick vectors. The aim of the present study was to investigate the prevalence of Ehrlichia sp. and Anaplasma sp. in 24 specimens of Azara's agouti (Dasyprocta azarae) that had been trapped in the Itapiracó Environmental Reserve, in São Luís, Maranhão, northeastern Brazil, using molecular methods. Four animals (16.7%) were positive for Ehrlichia spp. in nested PCR assays based on the 16S rRNA gene. In a phylogenetic analysis based on the 16S rRNA gene, using the maximum likelihood method and the GTRGAMMA+I evolutionary model, Ehrlichia sp. genotypes detected in Azara's agoutis were found to be closely related to E. canis and to genotypes relating to E. canis that had previously been detected in free-living animals in Brazil. The present work showed the first molecular detection of Ehrlichia sp. in Azara's agoutis in Brazil.

Molecular detection of Anaplasmataceae agents in Dasyprocta azarae in northeastern Brazil / Detecção molecular de agentes Anaplasmataceae em Dasyprocta azarae no nordeste do Brasil

Abstract Recently, the importance of wild-living rodents for maintenance of pathogens of the family Anaplasmataceae in the environment was investigated. These mammals play a role as reservoirs for these pathogens and act as hosts for the immature stages of tick vectors. The aim of the present study was to investigate the prevalence of Ehrlichia sp. and Anaplasma sp. in 24 specimens of Azara's agouti (Dasyprocta azarae) that had been trapped in the Itapiracó Environmental Reserve, in São Luís, Maranhão, northeastern Brazil, using molecular methods. Four animals (16.7%) were positive for Ehrlichia spp. in nested PCR assays based on the 16S rRNA gene. In a phylogenetic analysis based on the 16S rRNA gene, using the maximum likelihood method and the GTRGAMMA+I evolutionary model, Ehrlichia sp. genotypes detected in Azara's agoutis were found to be closely related to E. canis and to genotypes relating to E. canis that had previously been detected in free-living animals in Brazil. The present work showed the first molecular detection of Ehrlichia sp. in Azara's agoutis in Brazil.

Resumo Recentemente, a importância de roedores selvagens na manutenção de agentes Anaplasmataceae no ambiente tem sido investigada, haja visto o papel que tais mamíferos podem desempenhar como reservatórios para os patógenos e como hospedeiros para estágios imaturos dos carrapatos vetores. O presente estudo objetivou investigar a ocorrência de Ehrlichia sp. e Anaplasma sp. em 24 cotias (Dasyprocta azarae) capturadas na Reserva Ambiental de Itapiracó, em São Luís, Maranhão, nordeste do Brazil, utilizando métodos moleculares. Quatro animais (16,7%) mostraram-se positivos nos ensaios de nested PCR para Ehrlichia spp. baseados no gene 16S rRNA gene. Na análise filogenética baseda no gene 16S rRNA e utilizando o método de Máxima Verossimilhança e modelo evolutivo GTRGAMMA+I, os genótipos de Ehrlichia sp. detectados em cotias mostraram-se filogeneticamente relacionados às sequências de E. canis e outros genótipos relacionados a E. canis detectados previamente em animais selvagens no Brasil. O presente trabalho mostrou a primeira detecção molecular de Ehrlichia sp. em cotias no Brasil.

Genetic diversity of piroplasmids species in equids from island of São Luís, northeastern Brazil.

Equine piroplasmosisis, a tick-borne disease caused by the intra-erythrocytic protozoans Babesia caballi and Theileria equi, has economic importance due to the international trade and the increased movement of horses all over the world. The goal of this study was to evaluate the occurrence of phylogenetic diversity of T. equi and B. caballi genotypes among infected equids from São Luís Island, state of Maranhão, northeastern Brazil. Between December of 2011 and June of 2012, EDTA-blood and serum samples were collected from 139 equids (90 donkeys, 39 horses and 10 mules). From 139 serum samples submitted to ELISA assay, IgG antibodies to T. equi and B. caballi were detected in 19.4% (27/139) and 25.2% (35/139), respectively. Among sampled animals, 21.6% (30/139) and 55.4% (77/139) were positive for cPCR assays for T. equi and B. caballi, based on ema-1 and rap-1 genes, respectively. Overall, the T. equi sequences (n=7) submitted to Maximum Likelihood analysis (based on a 18S rRNA fragment of 1700 bp after alignment) grouped into three main groups, which were subdivided in eight clusters. The present work showed that different genotypes of T. equi and B. caballi circulate among equids in Brazil.

Genetic diversity of piroplasmids species in equids from island of São Luís, northeastern Brazil / Diversidade genética de espécies de piroplasmídeos em equídeos na ilha de São Luís, Nordeste do Brasil

Abstract Equine piroplasmosisis, a tick-borne disease caused by the intra-erythrocytic protozoans Babesia caballi and Theileria equi, has economic importance due to the international trade and the increased movement of horses all over the world. The goal of this study was to evaluate the occurrence of phylogenetic diversity of T. equi and B. caballi genotypes among infected equids from São Luís Island, state of Maranhão, northeastern Brazil. Between December of 2011 and June of 2012, EDTA-blood and serum samples were collected from 139 equids (90 donkeys, 39 horses and 10 mules). From 139 serum samples submitted to ELISA assay, IgG antibodies to T. equi and B. caballi were detected in 19.4% (27/139) and 25.2% (35/139), respectively. Among sampled animals, 21.6% (30/139) and 55.4% (77/139) were positive for cPCR assays for T. equi and B. caballi, based on ema-1 and rap-1 genes, respectively. Overall, the T. equi sequences (n=7) submitted to Maximum Likelihood analysis (based on a 18S rRNA fragment of 1700 bp after alignment) grouped into three main groups, which were subdivided in eight clusters. The present work showed that different genotypes of T. equi and B. caballi circulate among equids in Brazil.

Resumo A piroplasmose equina, uma doença transmitida por carrapatos e causada pelos protozoários intra-eritrocíticos Babesia caballi e Theileria equi, tem importância econômica devido ao comércio internacional e ao aumento do movimento de cavalos em todo o mundo. O objetivo do presente estudo foi mostrar a diversidade filogenética de T. equi e B. caballi infectando cavalos, burros e jumentos na Ilha de São Luís, Estado do Maranhão, Nordeste do Brasil. Entre dezembro de 2011 e junho de 2012, amostras de sangue com EDTA e soro de foram coletadas de 139 equídeos (90 jumentos, 39 cavalos e 10 burros). Dentre as 139 amostras de soro submetidas ao ensaio de ELISA, foram detectados anticorpos IgG contra T. equi e B. caballi em 19,4% (27/139) e 25,2% (35/139), respectivamente. Entre os animais amostrados, 21,6% (30/139) e 55,4% (77/139) foram positivos por meio dos ensaios de cPCR para T. equi e B. caballi, com base nos genes ema-1 e rap-1, respectivamente. No geral, as sequências T. equi (n = 7) submetidas à análise de Máxima Verossimilhança (baseada em um fragmento do 18S rRNA de 1700 pb, após o alinhamento) foram agrupadas em três grupos principais, os quais foram subdivididos em oito grupos. O presente trabalho mostrou que diferentes genótipos de T. equi e B. caballi circulam entre equídeos no Brasil.

Molecular characterisation of Bartonella species in cats from São Luís, state of Maranhão, north-eastern Brazil.

Bartonella species are fastidious bacteria that predominantly infect mammalian erythrocytes and endothelial cells and cause long-lasting bacteraemia in their reservoir hosts. Reports that describe the epidemiology of bartonellosis in Brazil are limited. This study aimed to detect and characterise Bartonella spp DNA from cat blood samples in São Luís, Maranhão, north-eastern Brazil. Among 200 cats tested for multiple genes, nine (4.5%) were positive for Bartonella spp: six cats for Bartonella henselae and three for Bartonella clarridgeiae. Based on the phylogenetic analysis of four genes, the B. henselae strain matched strains previously observed in Brazil and was positioned in the same clade as B. henselae isolates from the United States of America. Moreover, sequence alignment demonstrated that the B. clarridgeiae strain detected in the present study was the same as the one recently detected in cats from southern Brazil.

Molecular characterisation of Bartonella species in cats from São Luís, state of Maranhão, north-eastern Brazil

Bartonella species are fastidious bacteria that predominantly infect mammalian erythrocytes and endothelial cells and cause long-lasting bacteraemia in their reservoir hosts. Reports that describe the epidemiology of bartonellosis in Brazil are limited. This study aimed to detect and characterise Bartonella spp DNA from cat blood samples in São Luís, Maranhão, north-eastern Brazil. Among 200 cats tested for multiple genes, nine (4.5%) were positive for Bartonella spp: six cats for Bartonella henselae and three for Bartonella clarridgeiae. Based on the phylogenetic analysis of four genes, the B. henselae strain matched strains previously observed in Brazil and was positioned in the same clade as B. henselae isolates from the United States of America. Moreover, sequence alignment demonstrated that the B. clarridgeiae strain detected in the present study was the same as the one recently detected in cats from southern Brazil.

Occurrence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in cats with outdoor access in São Luís, Maranhão, Brazil.

The present study aimed to investigate the frequency of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in cats with outdoor access in São Luís, Maranhão, Brazil. The presence of IgG anti-T. gondii and anti-N. caninum antibodies was tested using the Indirect Immunofluorescent Antibody Test (IFAT). IgG anti-T. gondii and anti-N. caninum antibodies were detected in 101 (50.5%) and 54 (27%) sampled cats, respectively. The titers of anti-T. gondii antibodies ranged from 40 (cut-off) to 2560. On the other hand, the titers of anti-N. caninum antibodies ranged from 25 (cut-off) to 400. Twenty-seven cats (13.5%) were shown to be seropositive for both parasites. Seventy-four cats (34%) were seropositive only for T. gondii. Twenty-two cats (11%) were seropositive only for N. caninum. The present study showed that cats with outdoor access in São Luís, Maranhão, are exposed to T. gondii and N. caninum.

Occurrence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in cats with outdoor access in São Luís, Maranhão, Brazil / Ocorrência de anticorpos anti-Toxoplasma gondii e anti-Neospora caninum em gatos com acesso à rua em São Luís, Maranhão, Brasil

The present study aimed to investigate the frequency of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in cats with outdoor access in São Luís, Maranhão, Brazil. The presence of IgG anti-T. gondii and anti-N. caninum antibodies was tested using the Indirect Immunofluorescent Antibody Test (IFAT). IgG anti-T. gondii and anti-N. caninum antibodies were detected in 101 (50.5%) and 54 (27%) sampled cats, respectively. The titers of anti-T. gondii antibodies ranged from 40 (cut-off) to 2560. On the other hand, the titers of anti-N. caninum antibodies ranged from 25 (cut-off) to 400. Twenty-seven cats (13.5%) were shown to be seropositive for both parasites. Seventy-four cats (34%) were seropositive only for T. gondii. Twenty-two cats (11%) were seropositive only for N. caninum. The present study showed that cats with outdoor access in São Luís, Maranhão, are exposed to T. gondii and N. caninum.

O presente estudo objetivou verificar a frequência de anticorpos anti-Toxoplasma gondii e anti-Neospora caninum em gatos com acesso à rua em São Luís, Maranhão, Brasil. A presença de anticorpos IgG anti-T. gondii e anti-N. caninum foi verificado pela Reação de Imunofluorescência Indireta (RIFI). Anticorpos IgG anti-T. gondii e anti-N. caninum foram detectados em 101 (50,5%) e 54 (27%) gatos amostrados, respectivamente. Os títulos de anticorpos anti-T. gondii variaram de 40 (ponto de corte) a 2560. Por outro lado, anticorpos anti-N. caninum variaram de 25 (ponto de corte) a 400. Vinte e sete gatos (13,5%) mostraram-se soropositivos para ambos os parasitas. Setenta e quatro gatos (34%) foram soropositivos somente para T. gondii. Vinte e dois gatos (11%) foram soropositivos somente para N. caninum. O presente estudo demonstrou que gatos com acesso à rua em São Luís, Maranhão, são expostos ao T. gondii e N. caninum.

Molecular and serological detection of Ehrlichia spp. in cats on São Luís Island, Maranhão, Brazil.

Ehrlichiosis is a tick-borne disease that affects both humans and animals. The few existing reports on ehrlichiosis in Brazilian cats have been based on observation of morulae in leukocytes and, more recently, on molecular detection of Ehrlichia sp. In this study, we assessed occurrences of Ehrlichia sp. in the blood of 200 domestic cats in São Luís, Maranhão. Of the 200 animals tested, 11 (5.5%) were seropositive for Ehrlichia sp. and two (1%) were positive for Ehrlichia sp. in PCR. We also performed DNA sequence alignment to establish the identity of the parasite species infecting these animals, using the gene 16S rRNA. One cat presented infection with Ehrlichia sp. with 98% identity with E. canis, and another cat infected with Ehrlichia sp. showed 97% identity with E. chaffeensis. This is the first study on molecular detection of Ehrlichia sp. among domestic cats in São Luís, Maranhão.

Molecular and serological detection of Ehrlichia spp. in cats on São Luís Island, Maranhão, Brazil / Detecção molecular e sorológica de Ehrlichia spp. em gatos da ilha de São Luís, Maranhão, Brasil

Ehrlichiosis is a tick-borne disease that affects both humans and animals. The few existing reports on ehrlichiosis in Brazilian cats have been based on observation of morulae in leukocytes and, more recently, on molecular detection of Ehrlichia sp. In this study, we assessed occurrences of Ehrlichia sp. in the blood of 200 domestic cats in São Luís, Maranhão. Of the 200 animals tested, 11 (5.5%) were seropositive for Ehrlichia sp. and two (1%) were positive for Ehrlichia sp. in PCR. We also performed DNA sequence alignment to establish the identity of the parasite species infecting these animals, using the gene 16S rRNA. One cat presented infection with Ehrlichia sp. with 98% identity with E. canis, and another cat infected with Ehrlichia sp. showed 97% identity with E. chaffeensis. This is the first study on molecular detection of Ehrlichia sp. among domestic cats in São Luís, Maranhão.

Erliquiose é uma enfermidade transmitida por carrapatos que afeta seres humanos e animais. Os poucos relatos de erliquiose em gatos, no Brasil, são baseados na observação de mórulas em leucócitos e, mais recentemente, na detecção molecular de Ehrlichia sp. Neste estudo, foi avaliada a ocorrência de Ehrlichia sp. no sangue de 200 gatos de São Luís, Maranhão. Dos 200 animais testados, 11 (5,5%) foram soropositivos para Ehrlichia sp. e dois (1%) foram positivos na PCR para Ehrlichia spp. O alinhamento de sequências de DNA baseado no gene 16S rRNA foi conduzido para estabelecer a identidade da espécie de parasito que infectou estes animais. Um gato apresentou infecção por uma espécie de Ehrlichia sp. com 98% de identidade com E. canis; e outro mostrou-se infectado por Ehrlichia sp. com 97% de identidade com E. chaffeensis. Este estudo traz a primeira detecção molecular de Ehrlichia sp. em gatos de São Luís, Maranhão.