Levantamento sorológico e molecular de patógenos transmitidos por carrapatos em cães do Norte do Brasil

O estudo teve como objetivo avaliar a ocorrência de anticorpos anti-Rickettsia spp. e anti-Ehrlichia canis, bem como a detecção molecular de DNA de Babesia vogeli, Ehrlichia spp. e Hepatozoon spp. em cães do município de Ji-Paraná, estado de Rondônia, norte do Brasil. Para tanto, foram analisadas 177 amostras de soro e sangue por meio reação de imunofluorescência indireta (RIFI) e reação em cadeia pela polimerase (PCR), respectivamente. Um total de 8,47 (15/177) cães foram sororreativos para pelo menos uma espécie de Rickettsia, com títulos finais variando de 64 a 1.024. Os títulos finais mais elevados foram observados para Rickettsia parkeri e Rickettsia rhipicephali. Além disso, anticorpos anti-E. canis foram detectados em 24,29% (43/177) das amostras de soro testadas, com títulos finais variando de 80 a 2.560. Dentre as amostras de sangue analisadas molecularmente, 5,65% (10/177), 22,03% (39/177) e 48,02% (85/177) estavam infectados por B. vogeli, E. canis e Hepatozoon canis, respectivamente. Neste trabalho, fornecemos evidências sorológicas de Rickettsia spp., e E. canis além disso, confirmamos a ocorrência de DNA de B. vogeli, E. canis e H. canis em cães da área rural do estado de Rondônia. região oeste da Amazônia brasileira

Rural area of the Brazilian Pantanal wetlands associated with the occurrence of anti-Leishmania spp. antibodies in dogs

Neste trabalho foi realizada uma avaliação sobre a presença de anticorpos anti-Leishmania infantum chagasi em cães domésticos das áreas urbanas e rurais da região do Pantanal brasileiro usando técnicas sorológicas. Um total de 429 cães foram amostrados em três áreas do bioma do Pantanal, incluindo os municípios de Poconé, Santo Antônio de Leverger e Barão de Melgaço, em Mato Grosso, e o município de Corumbá, em Mato Grosso do Sul. A reação de imunofluorescência indireta (RIFI) foi utilizada para detectar anticorpos (ponto de corte de 40) de Leishmania infantum chagasi como antígeno. Devido à possibilidade de reação cruzada entre as espécies do gênero Leishmania, as amostras positivas na RIFI para L. infantum chagasi foram também avaliadas na RIFI utilizando L. amazonensis e L. braziliensis como antígenos. As amostras positivas na RIFI para L. infantum chagasi foram avaliadas utilizando o ensaio de imunoadsorção ligado à enzima (ELISA). Os resultados mostraram a presença de anticorpos contra L. infantum chagasi em 23 (5,36%; IC 95%: 3,50% -8,05%) cães e pelo menos um cão soro-reativo foi encontrado em todos os municípios avaliados neste estudo. Os títulos de anticorpos variaram de 40 a 5.120 e todas as amostras positivas na RIFI foram positivas no ELISA. Entre os 23 cães positivos, nove também reagiram para L. amazonensis e L. braziliensis. A ocorrência de anticorpos anti-L. infantum chagasi em cães foi maior nas áreas rurais (7,06%) do que nas áreas urbanas (2,50%) (P < 0,05). Com base neste estudo, concluímos que cães de áreas rurais do Pantanal tiveram contato com espécies de Leishmania, o que é uma informação relevante, dada a sua importância para a saúde pública.An evaluation was made of the presence of anti-Leishmania infantum chagasi antibodies in domestic dogs from the urban and rural areas of Brazil’s Pantanal wetland region using serological techniques. A total of 429 dogs were sampled in three areas of the Pantanal biome, including the municipalities of Poconé, Santo Antônio de Leverger, and Barão de Melgaço, in the state of Mato Grosso, and in the municipality of Corumbá, in the state of Mato Grosso do Sul. The immunofluorescence assay (IFA) was used to detect antibodies (cut-off point 40) using Leishmania infantum chagasi antigen. Because of the possibility of cross-reactivity between species of the genus Leishmania, samples that were positive in the IFA against L. infantum chagasi were also tested by IFA in the same conditions, using L. amazonensis and L. braziliensis antigens. IFA-positive samples to L. infantum chagasi were also evaluated using Enzyme-Linked Immunosorbent Assay (ELISA). The results showed the presence of antibodies against L. infantum chagasi in 23 (5.36%; 95% CI: 3.50%-8.05%) dogs and at least one seroreactive dog was found in each of the municipalities evaluated in this study. Antibody titers ranged from 40 to 5,120, and all IFA positive samples were positive in the ELISA. Among the 23 positive dogs, nine were also were seroreactive for L. amazonensis and L. braziliensis antigens. The occurrence of anti- L. infantum chagasi antibodies in dogs was higher in rural areas (7.06%) than in urban areas (2.50%) (P < 0.05). Based on this study, we concluded that dogs from rural areas of the Pantanal wetlands were in contact with Leishmania species, which is relevant information given their importance to public health

Detecção molecular e soroprevalência de Ehrlichia sp. em bovinos leiteiros da Amazônia ocidental, Brasil

Ehrlichia minasensis, uma espécie de Ehrlichia que está intimamente relacionada a E. canis, é conhecida por infectar bovinos e veados no Canadá. Esta bactéria rickettsial foi isolada de carrapatos Rhipicephalus (Boophilus) microplus e de bovino leiteiro do Sudeste e Centro-Oeste do Brasil, respectivamente. O objetivo deste estudo foi determinar a soroprevalência e realizar a detecção molecular de Ehrlichia sp. em bovinos leiteiros da região Norte do Brasil. O estudo foi realizado no município de Ji-Paraná, localizado no estado de Rondônia, região oeste da Amazônia brasileira. Amostras de sangue e soro foram obtidas entre setembro de 2012 a novembro de 2013 de vacas leiteiras (≥ 24 meses) em 64 fazendas. As amostras de sangue foram submetidas à reação em cadeia pela polimerase (PCR) para amplificar um fragmento do gene dsb de Ehrlichia sp. e os níveis de anticorpos de Ehrlichia sp. foram avaliados pelo teste de imunofluorescência indireta (IFI). Um total de 15 de 610 (2,45%; IC 95%: 1,04-3,86%) amostras de sangue apresentaram resultado positivo com base na detecção do gene dsb de Ehrlichia sp. O sequenciamento do produto da PCR confirmou que a sequência parcial amplificada do gene dsb (~ 295 pares de base) representava E. minasensis. Anticorpos anti-Ehrlichia sp. foram detectados em 178 animais (53,96%; IC 95%: 46,63-61,29%). Os títulos dos anticorpos variaram entre 40 e 5.120. No entanto, resultados positivos deste estudo devem ser interpretados com cautela. Entre as variáveis analisadas usando IFI, o número de vacas ≥ 24 meses foi estatisticamente significativo (p= 0,0103), e as propriedades com 51 a 100 vacas apresentaram maior probabilidade de infecção por Ehrlichia sp. Embora os animais não apresentem doença clínica, o caráter crônico da infecção pode levar à diminuição da produtividade.Ehrlichia minasensis, a novel species of Ehrlichia that is closely related to E. canis, is known to infect cattle and deer in Canada. This rickettsial bacterium was isolated from Rhipicephalus (Boophilus) microplus ticks and from dairy and beef cattle in southeast and midwestern Brazil, respectively. The purpose of this study was to determine the seroprevalence and perform molecular detection of Ehrlichia sp. in dairy cattle in the northern region of Brazil. The study was conducted in the municipality of Ji-Paraná, located in the state of Rondônia in the western Brazilian Amazon region. Blood and serum samples were obtained between the dates September 2012 and November 2013 from dairy cows (≥ 24 months old) in 64 farms. The blood samples were subjected to polymerase chain reaction (PCR) to amplify a fragment of the Ehrlichia sp. dsb gene, and the levels of Ehrlichia sp. antibodies were measured by the immunofluorescence assay (IFA). A total of 15 of 610 (2.45%; 95% CI: 1.04-3.86%) blood samples tested positive for ehrlichial infection based on the detection of the Ehrlichia sp. dsb gene. Sequencing of PCR amplicons from samples confirmed that the amplified partial dsb gene (~295 base pairs) sequence represented E. minasensis. Anti-Ehrlichia sp. antibodies were detected in 178 cows (53.96%; 95% CI: 46.63–61.29%). Endpoint titers ranged from 40 to 5,120. However, positive results derived from this assay should be interpreted with caution. Among the variables analyzed using IFA, the number of cows aged 24 months or greater was statistically significant (p = 0.0103), and hers of approximately 51-100 cows were more likely to be infected with Ehrlichia sp. Although the animals do not show clinical disease, the chronic character of the infection can lead to decrease in productivity

Infection by Neospora caninum in dairy cattle belonging to family farmers in the northern region of Brazil

Neosporosis is considered a major cause of abortion among cattle worldwide. The aim of this study was to investigate the prevalence of anti-Neospora caninum antibodies in dairy cattle and correlate them with possible risk factors on 63 small farms (family farms) in the municipality of Ji-Paraná, the main milk-producing region of the state of Rondônia, northern Brazil. For this purpose, 621 serum samples were collected from cows and were evaluated by means of the indirect fluorescence antibody test (IFAT). The overall herd prevalence of N. caninum antibodies among the farms (38/63; 60.31%) showed that N. caninum are widespread among the dairy herds in this region, despite only infecting a small proportion of animals (66/621, 10.62%). Occurrences of abortion and birth of weak calves were the only variables that showed as risk factors for the presence of N. caninum. The result from the spatial lag model strongly indicated that birth of weak calves and presence of N. caninum are occurring on farms that are located close to each other, indicating aggregation of disease occurrence

Seroprevalence, spatial analysis and risk factors of infection with Neospora caninum in cattle in Brazil’s northern Pantanal wetland

Abstract Neosporosis is caused by an obligate intracellular protozoan, Neospora caninum . It is considered one of the most widespread and frequent causes of abortion in cattle worldwide. To evaluate the prevalence of anti-N. caninum antibodies and associated risk factors, serum samples were collected from 2,452 bovines at 262 farms in the northern Pantanal, state of Mato Grosso, Brazil. Each farmer was asked to fill out a questionnaire for subsequent epidemiological data analysis. Anti-N. caninum antibodies were detected by means of the indirect immunofluorescent assay (IFA), using a cut-off dilution of 1:100. The overall anti-N. caninum antibodies prevalence was 25.44% (Confidence Interval - CI 95%; 20.10%; 30.78%), and the anti-N. caninum antibodies prevalence per herd was 76.72% (CI 95%; 71.60%; 81.84%). The presence of dogs, occurrence of abortion in cows, and sale of cattle for breeding were statistically associated with seropositivity in herds, while the risk of females being seropositive for N. caninum was higher in animals ≤ 6-years-old and in the presence of dogs. A spatial analysis indicated that the relative risk of the disease is spatially constant and that the farms with the highest prevalence of anti-N. caninum antibodies are located south of the region under study

Ocorrência de anticorpos anti-Neospora caninum em fêmeas e fetos da espécie bovina do município de Nossa Senhora do Livramento, Mato Grosso

Neosporosis is a parasitic disease caused by the obligate intracellular protozoan Neospora caninum, which is responsible for miscarriage cases in cattle countrywide. The vertical or transplacental route with prevalent miscarriages between the 3rd and 6rd months of pregnancyis the main transmission form in cattle. The aim of the present study is to investigate the presence of anti-N.caninum antibodies in (Bos indicus) Nellore female animals and fetuses in Nossa Senhora do Livramento County - MT. Blood samples from 489 female bovines and from 128 bovine fetuses over three months of pregnancy were colleted. The research about the anti N. caninum antibodies in bovine females and fetuses was conducted through the Immunofluorescence technique (IFA). We found 2.45% (12) seropositive and 3.91% (5) seropositive to N. caninum in the present study.A neosporose é uma doença parasitária ocasionada pelo protozoário intracelular obrigatório Neospora caninum, responsável por quadros de abortos em bovinos em várias regiões do país. A principal forma de transmissão entre os bovinos é a via vertical ou transplacentária com abortos prevalentes entre 3 a 6 meses de gestação. Este trabalho teve como objetivo pesquisar a presença de anticorpos anti-N.caninum em fêmeas e fetos bovinos da raça Nelore (Bos indicus), do município de Nossa Senhora do Livramento-MT. Durante o abate dos animais foram coletadas amostras de sangue de 489 fêmeas e de 128 fetos acima de três meses de gestação. A pesquisa de anticorpos anti N. caninum nas fêmeas e fetos foi determinada pela Reação de Imunofluorescência Indireta (RIFI). A ocorrência foi de 2,45% para as fêmeas e de 3,91% para os fetos

Genetic diversity and molecular survey of Trypanosoma (Megatrypanum ) theileri in cattle in Brazil’s western Amazon region

Abstract Trypanosoma (Megatrypanum) theileri is a flagellated protozoan that infects ruminants and it displays high genetic diversity. In this study, we investigated the prevalence rates of this protozoan based on hemoculture and molecular diagnosis. The isolates of T. theileri thus obtained were characterized by molecular markers SSU rDNA and gGAPDH and molecular diagnosis based on Cathepsin L-like gene (PCR-TthCATL). The PCR-TthCATL and hemoculture indicated an overall prevalence rate of 8.13%, and the CATL derived sequence named IB was identified for the first time in cattle in the western Amazon region, as well as IF in Brazil. We also describe a possible new PCR-TthCATL derived sequence in cattle, designated IL

Prevalence of canine monocitic ehrlichiosis and canine thrombocytic anaplasmosis in dogs suspected of hemoparasitosis in Cuiabá Mato Grosso

The present study evaluated Canine Monocytic Ehrlichiosis and Canine Thrombocytic Anaplasmosis in 77 dogs treated at the Veterinary Hospital (HOVET) of the Federal University of Mato Grosso (UFMT) in 2009. An association among clinical and hematological, serological and molecular data was investigated. Dogs were evaluated by partial amplification of dsb and 16S rRNA of Ehrlichia canis and Anaplasma platys and by Indirect Fluorescence Antigen Test (IFAT) for E. canis. DNA of E. canis and A. platys were detected in 18 (23.3%) and 07 (9.1%) dogs respectively. Fifty four (70.1%) dogs were positive by IFAT with titers ranging from 40 to 327,680. There were higher frequencies of E. canis positive PCR in dogs less than 12 months old, with anemia, lymphopenia and thrombocytopenia (P?0.05). Leukopenic dogs presented tendency to be A. platys PCR positive (P=0.07). Thrombocytopenia and hyperproteinemia of dogs were associated with high anti-E. canis antibody titers (P?0.05). Lymphadenopathy and pulmonary changes were observed in 15 (30.6%) and 4 (57.1%) E. canis PCR positive dogs (P?0.05). No other clinical features were associated between E. canis and A. platys infected dogs. E. canis was the only specie of Ehrlichia detected in dogs attended at the HOVET with a higher infection rate in young dogs, and statistically associated with anemic and thrombocytopenic dogs. On the other side, A. platys presented low occurrence among evaluated dogs

Prevalence of 'Candidatus Mycoplasma haemobos' detected by PCR, in dairy cattle from Ji-Paraná in the north region of Brazil

ABSTRACT: The municipality of Ji-Paraná, Rondônia, is one of the major dairy production areas in the north region of Brazil. Thus, it is important to evaluate infectious agents that have the potential to negatively affect productivity in the industry. The objective of this study was to investigate the prevalence of ' Candidatus Mycoplasma haemobos' by using a PCR-based detection method and correlate this with dairy herd variables (abortion frequency, weak calf birth rate, total cattle number, >24-month-old cow number, farm size, and production system) in family farms of the Ji-Paraná municipality, north region, Brazil. Blood samples were collected from 320 dairy cows located across 64 farms (i.e., five animals per farm) from September 2012 to November 2013. Overall prevalence of 'Ca. M. haemobos' was 64.2% and prevalence per herd was 95.3%; the number of >24-month-old cows in the farms studied correlated with ' Ca. M. haemobos' infection rates. Considering the importance of the dairy industry to the study area, additional investigations are necessary to evaluate the effect of chronic infection in these animals on milk production and herd health