Implications of domestic dogs in the epidemiology of Rickettsia parkeri strain Atlantic rainforest and Rangelia vitalii in Southeastern Brazil.

This study aimed to evaluate the occurrence of diseases transmitted by Amblyomma ovale in 61 dogs monitored for three years through collections of ticks and blood, interviews, telemetry and camera traps in three areas of Serra do Mar State Park, Brazil. Blood samples were used to investigate infection by Rangelia vitalii by real-time TaqMan PCR and Rickettsia parkeri by IIFA. The collected ticks were submitted to conventional PCR to investigate the presence of R. parkeri . These data were compared with the monitoring results and interviews with the owners. Dogs considered as companion presented a risk of infection by R. parkeri strain Mata Atlantica 5.4 times higher than those not considered as companion (p = 0.009). Dogs that had at least one A. ovale collected during the campaigns had a 10 times higher risk of infection by R. parkeri strain Mata Atlantica than those who did not (p = 0.009). One dog positive for R. vitalii by real-time TaqMan PCR was parasitized by A. ovale frequently during monitoring. Sequenced ompaA - positive DNA samples had 100% identity of R. parkeri strain Mata Atlantica clone As106. From the findings, it is urgent to control domestic dogs around rainforests to reduce zoonoses transmission.

Implications of domestic dogs in the epidemiology of Rickettsia parkeri strain Atlantic rainforest and Rangelia vitalii in Southeastern Brazil / Implicações dos cães domésticos na epidemiologia da Rickettsia parkeri cepa Mata Atlântica e da Rangelia vitalii no Sudeste do Brasil

Abstract This study aimed to evaluate the occurrence of diseases transmitted by Amblyomma ovale in 61 dogs monitored for three years through collections of ticks and blood, interviews, telemetry and camera traps in three areas of Serra do Mar State Park, Brazil. Blood samples were used to investigate infection by Rangelia vitalii by real-time TaqMan PCR and Rickettsia parkeri by IIFA. The collected ticks were submitted to conventional PCR to investigate the presence of R. parkeri . These data were compared with the monitoring results and interviews with the owners. Dogs considered as companion presented a risk of infection by R. parkeri strain Mata Atlantica 5.4 times higher than those not considered as companion (p = 0.009). Dogs that had at least one A. ovale collected during the campaigns had a 10 times higher risk of infection by R. parkeri strain Mata Atlantica than those who did not (p = 0.009). One dog positive for R. vitalii by real-time TaqMan PCR was parasitized by A. ovale frequently during monitoring. Sequenced ompaA - positive DNA samples had 100% identity of R. parkeri strain Mata Atlantica clone As106. From the findings, it is urgent to control domestic dogs around rainforests to reduce zoonoses transmission.

Resumo A ocorrência de doenças transmitidas por Amblyomma ovale em 61 cães monitorados por três anos através de coletas de carrapatos, sangue, entrevistas, telemetria e armadilhas fotográficas foi avaliada em três áreas do Parque Estadual da Serra do Mar - SP. Amostras de sangue foram utilizadas para investigação de Rangelia vitalii através de PCR TaqMan em tempo real e Rickettsia parkeri através da RIFI. Carrapatos coletados foram submetidos à PCR convencional para investigação de R. parkeri . Estes dados foram comparados considerando os resultados do monitoramento e entrevistas. Cães de companhia apresentaram risco de infecção pela R. parkeri cepa Mata Atlântica 5,4 vezes maior que os não considerados como de companhia (p = 0,009). Cães que tiveram pelo menos um A. ovale coletado apresentaram risco de infecção por R. parkeri cepa Mata Atlântica 10 vezes maior do que aqueles que não tiveram (p = 0,009). Um cão positivo para R. vitalii através de PCR TaqMan em tempo real foi parasitado por A. ovale durante o monitoramento. Amostras positivas para o gene ompaA possuíam 100% de identidade do clone As106 de R. parkeri cepa de Mata Atlântica. Assim, é urgente o controle de cães na Mata Atlântica para redução dos riscos de zoonoses.

Occurrence of Ehrlichia canis and Hepatozoon canis and probable exposure to Rickettsia amblyommatis in dogs and cats in Natal, RN / Ocorrência de Ehrlichia canis, Hepatozoon canis e provável exposição a Rickettsia amblyommatis em cães e gatos de Natal, RN

Abstract Occurrence of infection or exposure to Ehrlichia canis, Hepatozoon canis and Rickettsia spp. was detected in feral cats living in two fragments from Atlantic rainforest, in Natal, RN, Brazil, and in dogs living around the parks. While serum samples were collected from 155 animals (53 cats living in the parks; 29 dogs living in human homes around the parks; and 73 dogs living at an animal control center - ACC), spleen samples were collected from 20 dogs that were euthanized at ACC. Serum samples were analyzed to Rickettsia spp. and E. canis antibodies using the indirect immunofluorescence assay. Seventeen of the 102 dogs (17%) had E. canis antibodies and 13% (20/155) of all dogs and cats (i.e. 3% (3/102) of the dogs and 32% (17/53) of the cats) were seropositive for Rickettsia spp. antigens. The animals were therefore been exposed to R. amblyommatis or by a very closely related genotype. Among the 20 dog spleen samples analyzed, eight were PCR positive for E. canis and two for H. canis (GenBank accession number MG772657 and MG772658, respectively). In none of the spleen samples were obtained amplicons for Babesia spp. through PCR. This study provided the first evidence that Rickettsia of the spotted fever group is circulating among dogs and cats in Natal.

Resumo A ocorrência de infecção ou exposição para Ehrlichia canis, Hepatozoon canis e Rickettsia spp. foi determinada em gatos ferais que viviam em dois fragmentos da Mata Atlântica, localizados em Natal, RN, Brasil e em cães que viviam em torno dos parques e em outras regiões da cidade. Enquanto amostras de soro foram coletadas de 155 animais (53 gatos que viviam nos parques, 29 cães com domicilio em torno dos parques e 73 cães do Centro de Controle de Animais -CCA), fragmentos de baço foram coletados de 20 cães eutanasiados no CCA. A detecção de anticorpos nas amostras de soros coletadas contra Rickettsia spp. e E. canis foi realizada pela Reação de Imunofluorescência Indireta. Dezessete dos 102 cães (17%) apresentaram anticorpos anti E. canis e 13% (20/155) de todos os cães e gatos (ou seja, 3% (3/102) dos cães e 32% (17/53) dos gatos) foram soropositivos para antígenos de Rickettsia spp. Os animais foram considerados expostos à R. amblyommatis ou a um genótipo muito relacionado. Entre as 20 amostras de baço de cães analisadas, oito foram positivas para E. canis e duas para Hepatozoon canis (números de acesso ao Genbank MG772657 e MG772658, respectivamente). Nenhuma das amostras de baço produziram amplicons de Babesia spp. na PCR. Observou-se, pela primeira vez, a circulação de Rickettsia do grupo da febre maculosa em cães e gatos em Natal, RN.

Occurrence of Ehrlichia canis and Hepatozoon canis and probable exposure to Rickettsia amblyommatis in dogs and cats in Natal, RN.

Occurrence of infection or exposure to Ehrlichia canis, Hepatozoon canis and Rickettsia spp. was detected in feral cats living in two fragments from Atlantic rainforest, in Natal, RN, Brazil, and in dogs living around the parks. While serum samples were collected from 155 animals (53 cats living in the parks; 29 dogs living in human homes around the parks; and 73 dogs living at an animal control center - ACC), spleen samples were collected from 20 dogs that were euthanized at ACC. Serum samples were analyzed to Rickettsia spp. and E. canis antibodies using the indirect immunofluorescence assay. Seventeen of the 102 dogs (17%) had E. canis antibodies and 13% (20/155) of all dogs and cats (i.e. 3% (3/102) of the dogs and 32% (17/53) of the cats) were seropositive for Rickettsia spp. antigens. The animals were therefore been exposed to R. amblyommatis or by a very closely related genotype. Among the 20 dog spleen samples analyzed, eight were PCR positive for E. canis and two for H. canis (GenBank accession number MG772657 and MG772658, respectively). In none of the spleen samples were obtained amplicons for Babesia spp. through PCR. This study provided the first evidence that Rickettsia of the spotted fever group is circulating among dogs and cats in Natal.

Occurrence of Ehrlichia canis and Hepatozoon canis and probable exposure to Rickettsia amblyommatis in dogs and cats in Natal, RN

Abstract Occurrence of infection or exposure to Ehrlichia canis, Hepatozoon canis and Rickettsia spp. was detected in feral cats living in two fragments from Atlantic rainforest, in Natal, RN, Brazil, and in dogs living around the parks. While serum samples were collected from 155 animals (53 cats living in the parks; 29 dogs living in human homes around the parks; and 73 dogs living at an animal control center - ACC), spleen samples were collected from 20 dogs that were euthanized at ACC. Serum samples were analyzed to Rickettsia spp. and E. canis antibodies using the indirect immunofluorescence assay. Seventeen of the 102 dogs (17%) had E. canis antibodies and 13% (20/155) of all dogs and cats (i.e. 3% (3/102) of the dogs and 32% (17/53) of the cats) were seropositive for Rickettsia spp. antigens. The animals were therefore been exposed to R. amblyommatis or by a very closely related genotype. Among the 20 dog spleen samples analyzed, eight were PCR positive for E. canis and two for H. canis (GenBank accession number MG772657 and MG772658, respectively). In none of the spleen samples were obtained amplicons for Babesia spp. through PCR. This study provided the first evidence that Rickettsia of the spotted fever group is circulating among dogs and cats in Natal.

Resumo A ocorrência de infecção ou exposição para Ehrlichia canis, Hepatozoon canis e Rickettsia spp. foi determinada em gatos ferais que viviam em dois fragmentos da Mata Atlântica, localizados em Natal, RN, Brasil e em cães que viviam em torno dos parques e em outras regiões da cidade. Enquanto amostras de soro foram coletadas de 155 animais (53 gatos que viviam nos parques, 29 cães com domicilio em torno dos parques e 73 cães do Centro de Controle de Animais -CCA), fragmentos de baço foram coletados de 20 cães eutanasiados no CCA. A detecção de anticorpos nas amostras de soros coletadas contra Rickettsia spp. e E. canis foi realizada pela Reação de Imunofluorescência Indireta. Dezessete dos 102 cães (17%) apresentaram anticorpos anti E. canis e 13% (20/155) de todos os cães e gatos (ou seja, 3% (3/102) dos cães e 32% (17/53) dos gatos) foram soropositivos para antígenos de Rickettsia spp. Os animais foram considerados expostos à R. amblyommatis ou a um genótipo muito relacionado. Entre as 20 amostras de baço de cães analisadas, oito foram positivas para E. canis e duas para Hepatozoon canis (números de acesso ao Genbank MG772657 e MG772658, respectivamente). Nenhuma das amostras de baço produziram amplicons de Babesia spp. na PCR. Observou-se, pela primeira vez, a circulação de Rickettsia do grupo da febre maculosa em cães e gatos em Natal, RN.

Toxoplasma gondii in domestic and wild animals from forest fragments of the municipality of Natal, northeastern Brazil.

Toxoplasmosis stands out as a global disease that has felines as definitive hosts. In the municipality of Natal, Rio Grande do Norte State, Brazil, two parks are notable for their ecological and social importance. This study aimed to investigate the presence of Toxoplasma gondii in short hair cats, bats and small non-volant mammals in these two ecological reserves. Altogether, biological samples were obtained from 154 mammals, 92 wild animals from both areas and 62 domestic cats of the Parque da Cidade. In total, 22 (53.7%) non-volant wild mammals, 11 (21.5%) bats and 28 (52.8%) cats were positive for IgG anti-T. gondii antibodies using the Modified Agglutination Test (≥ 25). It was possible to detect the presence of T. gondii DNA, by means of a molecular amplification of a B1 gene fragment (155bp), in 92 tissue samples from wild animals, including Didelphis albiventris, Monodelphis domestica, Artibeus lituratus, Carollia perspicillata and Glossophaga soricina. Of the 62 cats examined by the same molecular method, T. gondii DNA could be detected in 4 cats. In this study, it was observed the circulation of T. gondii in wild species and domestic cats, demonstrating the involvement of wild and domestic animals in the cycle of T. gondii.

Toxoplasma gondii in domestic and wild animals from forest fragments of the municipality of Natal, northeastern Brazil / Toxoplasma gondii em animais domésticos e silvestres de fragmentos florestais do município de Natal, nordeste do Brasil

Toxoplasmosis stands out as a global disease that has felines as definitive hosts. In the municipality of Natal, Rio Grande do Norte State, Brazil, two parks are notable for their ecological and social importance. This study aimed to investigate the presence of Toxoplasma gondii in short hair cats, bats and small non-volant mammals in these two ecological reserves. Altogether, biological samples were obtained from 154 mammals, 92 wild animals from both areas and 62 domestic cats of the Parque da Cidade. In total, 22 (53.7%) non-volant wild mammals, 11 (21.5%) bats and 28 (52.8%) cats were positive for IgG anti-T. gondii antibodies using the Modified Agglutination Test (≥ 25). It was possible to detect the presence of T. gondii DNA, by means of a molecular amplification of a B1 gene fragment (155bp), in 92 tissue samples from wild animals, including Didelphis albiventris, Monodelphis domestica, Artibeus lituratus, Carollia perspicillata and Glossophaga soricina. Of the 62 cats examined by the same molecular method, T. gondii DNA could be detected in 4 cats. In this study, it was observed the circulation of T. gondii in wild species and domestic cats, demonstrating the involvement of wild and domestic animals in the cycle of T. gondii.

Toxoplasmose destaca-se como uma doença global que tem felinos como hospedeiros definitivos. No município de Natal, Rio Grande do Norte, Brasil, dois parques são notáveis por sua importância ecológica e social. Este estudo teve como objetivo investigar a presença de Toxoplasma gondii em gatos de rua, morcegos e pequenos mamíferos não-voadores nestas duas reservas ecológicas. No total, as amostras biológicas foram obtidas de 154 mamíferos, 92 animais selvagens de ambas as áreas e 62 gatos domésticos do Parque da Cidade. No total, 22 (53,7%) mamíferos silvestres não-voadores, 11 (21,5%) morcegos e 28 (52,8%) gatos foram positivos para IgG anti-T. gondii utilizando o Teste de Aglutinação Modificado (≥ 25). Foi possível detectar a presença de DNA de T. gondii, por meio de uma amplificação molecular de um fragmento do gene B1 (155bp), em 92 amostras de tecido de animais selvagens, incluindo Didelphis albiventris, Monodelphis domestica, Artibeus lituratus, Carollia perspicillata e Glossophaga soricina. Dos 62 gatos examinadas pelo mesmo método molecular, DNA de T. gondii pode ser detectado em quatro gatos. Neste estudo, observou-se a circulação de T. gondii em espécies selvagens e gatos domésticos, demonstrando o envolvimento de animais domésticos e selvagens no ciclo de T. gondii.

Toxoplasma gondii: evaluation of immunofluorescence assay using heterologous secondary antibody in experimentally infected wild small rodents / Toxoplasma gondii: avaliação da reação de imunofluorescência indireta usando anticorpo secundário heterólogo em roedores silvestres experimentalmente infectados

Toxoplasmosis is a zoonotic infection caused by Toxoplasma gondii that affects a wide range of vertebrates. Rodents are intermediate hosts and serve as food for felids, the definitive hosts. However, because of the high variety of species in the order Rodentia, the serological diagnosis is difficult to perform, since the most used techniques require the use of specific antibody conjugated to fluorescein or enzymes. The aim of this study was to evaluate the use of a heterologous secondary antibody conjugate in Immunofluorescence Antibody Test (IFAT) for diagnosis of anti-T. gondii antibodies in two species of wild rodents: Euryoryzomys russatus (WAGNER, 1848) and Calomys callosus (RENGGER, 1830). The specie Mus musculus (Linnaeus, 1758) was used as control conjugate. The animals were experimentally infected with five cysts of T. gondii (strain ME 49) per animal. The Modified Agglutination Test (MAT), which does not require the use of conjugates, and the presence of T. gondii cysts in the rodents were used to confirm the infection. For each animal species, serum samples were collected weekly for five weeks and tested (50 samples per rodent specie, total of 150 samples). None of the samples from C. callosus and E. russatus were positive in the IFAT when anti-mouse heterologous conjugate was used. Brain cysts of T. gondii were microscopically observed in all animals, except in one of the E. russatus. Positive results were found in the MAT 14 days after T. gondii infection in all three species of rodents and IFAT of the control group (M. musculus) was also positive 14 days after infection using anti-mouse (homologous) conjugate. The use of heterologous secondary antibody conjugates should be used with caution and the MAT had a good agreement for serological diagnosis of T. gondii in the studied rodent species.

A toxoplasmose é uma zoonose causada pelo Toxoplasma gondii, que afeta uma grande variedade de vertebrados. Os roedores são hospedeiros intermediários e servem de alimento para felinos, os hospedeiros definitivos. No entanto, por causa da elevada variedade de espécies na ordem Rodentia, o diagnóstico serológico é difícil de ser realizado, uma vez que as técnicas mais empregadas requerem a utilização de um anticorpo específico conjugado com fluoresceína ou enzimas. O objetivo deste estudo foi avaliar o uso de um conjugado secundário heterólogo na Reação de Imunofluorescência Indireta (RIFI) para diagnóstico de anticorpos anti-T. gondii em duas espécies de roedores silvestres: Euryoryzomys russatus (WAGNER, 1848) e Calomys callosus (RENGGER, 1830). A espécie Mus musculus (Linnaeus, 1758) foi utilizada como controle do conjugado. As três espécies de roedores foram experimentalmente infectadas com cinco cistos de T. gondii (cepas ME 49) por animal. O Teste de Aglutinação Modificado (MAT), que não requer o uso de conjugados, bem como, a presença de cistos de T. gondii nos roedores foram usados para confirmar a infecção. Para cada espécie de animal, amostras de soro foram coletadas durante cinco semanas e testadas (50 amostras por espécie de roedor, total de 150 amostras). Nenhuma das amostras de C. callosus e E. russatus foram positivas na RIFI, quando foi usado o conjugado heterólogo anti-camundongo. Cistos cerebrais de T. gondii foram microscopicamente observados em todos os animais, exceto em um dos E. russatus. Resultados positivos foram encontrados pelo MAT após 14 dias de inoculação em todas as três espécies estudadas e pela RIFI no grupo controle (M. musculus), também no dia 14 após a infecção utilizando conjugado anti-camundongo. O uso de conjugados secundários heterólogos deve ser empregado com cautela e o MAT apresentou uma boa concordância para o diagnóstico sorológico de T. gondii nas espécies de roedores estudadas.