Costa Rican Genotype of Ehrlichia canis: A Current Concern.

Canine monocytic ehrlichiosis (CME) is endemic to Brazil, and studies have verified that dogs have been exposed to different genotypes of Ehrlichia canis. This genetic divergence can influence the clinical response of the animals. We aimed to describe clinical and hematological changes in 125 dogs that reacted to BrTRP36, USTRP36, and CRTRP36 genotypes through enzyme immunoassays and to highlight the current concern regarding infection by the Costa Rican genotype. The results showed that 52.0% reacted to the Brazilian genotype, 22.4% reacted to the Costa Rican genotype, and 16.0% reacted to the American genotype, and some co-reactions were observed. Dogs reactive to BrTRP36 were 1.24% more likely to present with medullary regeneration in cases of anemia and 3% less likely to manifest hyperproteinemia, while dogs reactive to CRTRP36 were 0.7% less likely to present with medullary regeneration. Febrile illness and neurological alterations were also statistically associated, with an 85.7% and 231.2% increased likelihood, respectively, to occur in dogs that reacted to USTRP36. The dogs with the American genotype developed clinical manifestations related to systemic inflammation, while those with the Brazilian genotype of E. canis were more dispersed in the region studied, showing greater adaptation to the hosts. We highlight the significant serocurrence of the Costa Rican genotype, which has already been described to have zoonotic potential and which showed less adaptation.

Diversity of rickettsiae in ticks (Acari: Ixodidae) collected from wild vertebrates in part of the Amazon, Cerrado, and Pantanal biomes in Brazil.

Ticks parasitizing 102 wild animals in the states of Mato Grosso and Goiás, Brazil were collected between 2015 and 2018. A total of 2338 ticks (865 males, 541 females, 823 nymphs, and 109 larvae) belonging to four genera (Amblyomma, Dermacentor, Haemaphysalis, and Rhipicephalus) and at least 21 species were identified. DNA extraction and a molecular survey for rickettsial agents were performed on 650 ticks. The results revealed parasitism by the following species: Rickettsia amblyommatis in Amblyomma cajennense s.s., A. cajennense s.l., Amblyomma coelebs, Amblyomma humerale, Amblyomma longirostre, Amblyomma nodosum, Amblyomma scalpturatum, Amblyomma sculptum, and Amblyomma romitii; Rickettsia parkeri in Amblyomma nodosum, Amblyomma ovale, Amblyomma scalpturatum, and Amblyomma triste; Rickettsia rhipicephali in Haemaphysalis juxtakochi; Rickettsia sp. in A. cajennense s.s., A. nodosum, and A. sculptum, and lastly, 'Candidatus Rickettsia andeanae' in Amblyomma parvum and Rhipicephalus microplus. This study expands the body of knowledge about tick parasitism among wild animals, including new data concerning tick-host associations, and provides information about the epidemiology of tick-borne pathogens in the Center-West region of Brazil.

Seroprevalence of Toxoplasma gondii and Neospora caninum in sheep intended for human consumption in the Rondônia state, Western Brazilian Amazon.

The aim of this study was to carry out a seroepidemiological study of anti-T. gondii and anti-N. caninum antibodies in serum samples from sheep intended for human consumption in the Rondônia state, in the Western Brazilian Amazon, and identify possible risk factors associated with seropositivity. The analysis of the 616 sheep serum samples revealed animal-level seroprevalence of T. gondii in the order of 52.4 % and of N. caninum of 60.6 %. Prevalence of co-infections was 33.4 %. The herd-level seroprevalence was 100 % for both the parasites. Applying of multivariate logistic regression analysis, test of the association between seroprevalence of T. gondii infection and the potential predictors showed that feed, fetal malformation, abortion, neurological problems (lambs), presence of rodents were the significant (P < 0.05) predictors. The variables presence of rodents, cats, wild animals and main activity showed statistical association (P < 0.05) with the seroprevalence of N. caninum. Due to the high prevalence of T. gondii and N. caninum found in this study, the absence of specific slaughterhouses for sheep and raw or undercooked sheep meat for human consumption we concluded that sheep can represent an important source of infection for humans.

Antibodies anti-Toxoplasma gondii and anti-Neospora caninum in backyard pigs from the state of Mato Grosso, Brazil.

To estimate the seroprevalence of Toxoplasma gondii and Neospora caninum, using an indirect immunofluorescent assay (IFA), and identify the risk factors associated, serum samples were collected from 1,070 pigs from 320 backyard pig farming in the of Mato Grosso state. The animal-level seroprevalence of T. gondii and N. caninum was 32.48% and 13.49%, respectively, with a herd seroprevalence of 55.63% for T. gondii and 27.81% for N. caninum. Feeding the animals with leftovers increases the probability of the presence of anti-T. gondii antibodies in pigs by 1.09-fold. Unlike to T. gondii, feeding with leftovers was found to be negatively associated with N. caninum seropositivity in farm-level analysis and in the animal-level model, so decreasing the chances of positivity. Yet, age was considered a risk factor for N. caninum seropositivity. Further studies are necessary to evaluate the impact of T. gondii infection on backyard pig farming production, and its importance as a source of toxoplasmosis infection in humans in the Mato Grosso state, as well as, the role of domestic pigs in the epidemiology of neosporosis.

Molecular survey of tick-borne pathogens in small mammals from Brazilian Amazonia.

Small non-volant mammals (marsupials and small rodents) were captured at three different timepoints from 23 forest fragments across three municipalities (Alta Floresta, Sinop and Cláudia) covering the Amazonian biome of the Mato Grosso State in Midwestern Brazil. The animal tissues (liver and spleen) and blood were screened using molecular tools for the detection of Babesia, Coxiella, Cytauxzoon, Hepatozoon, Theileria, and Anaplasmataceae agents. A total of 230 specimens (78 rodents and 152 marsupials) were trapped. Hepatozoon and Piroplasmorida agents were detected in the common opossums (Didelphis marsupialis). In turn, all samples (blood, liver, or spleen) collected from the small mammals were negative for the genus Coxiella and the family Anaplasmataceae, as detected by polymerase chain reaction (PCR). Phylogenetic analyses inferred from partial sequences of the 18S rRNA gene highlighted the occurrence of new Hepatozoon and Piroplasmorida haplotypes. Future studies determining the role of common opossum (D. marsupialis) in the epidemiological cycles of Hepatozoon and Babesia under natural conditions in the Amazonian biome are necessary.

Detection molecular of Rangelia vitalii in dogs from Parana State, Southern Brazil.

Rangelia vitalii infects erythrocytes, leukocytes and endothelial cells of dogs. The present study aimed to report the molecular detection confirmed by sequencing of R. vitalii in the state of Paraná, as well as describe the clinical, hematological and biochemical alterations of the infected dogs. Three sick dogs from the metropolitan area of Curitiba, PR, Brazil, underwent a physical exam, and laboratory tests included hematology, biochemistry, polymerase chain reaction (PCR), and gene sequencing. Clinical signs included apathy, anorexia, and hemorrhage. Intra-erythrocytic and extracellular piroplasms were found on peripheral blood smears from all three dogs. Blood samples from these animals were positive for Babesia sp. by PCR targeting 18S rRNA. PCR products from all three dogs were sequenced, and BLAST analysis showed that the PCR-generated sequences were highly homologous with those of R. vitalii previously reported. Hematologic findings included severe anemia, shift of neutrophils to the regenerative left, and thrombocytopenia. Serum urea levels were increased in all three dogs, and direct bilirubin levels were elevated in one dog.

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

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.

Toxoplasma gondii exposure in felines: an alarming factor to public health from mineiros, Goiás / Exposição ao toxoplasma gondii em felinos: um fator alarmante para a saúde pública de Mineiros, Goiás / Exposición a toxoplasma gondii en felinos: un factor alarmante para la salud pública de Mineiros, Goiás

Toxoplasmosis is a cosmopolitan disease caused by Toxoplasma gondii, an obligate protozoan that affects endothermic animals, including man. Being a zoonosis that can cause abortions, fetal malformations and blindness. Cats are the definitive hosts, acting as a source of dissemination. In order to collect epidemiological data on the distribution of T. gondii in Mineiros, Goiás, samples of domestic cats were analyzed to verify the circulation of the protozoan and reinforce preventive measures. Ninety-four samples of domestic cats, regardless of gender, age and breed, were randomly processed in the neighborhoods of Mineiros, Goiás. They were submitted to the indirect fluorescent antibody test for antibodies against T. gondii, using tachyzoites as antigens and anti-cat IgG conjugate (1:16 were positive). Of the 94 samples analyzed, 35 (37.2%) were reagents, of which four had a titration of 1:16, ten of 1:32, eleven of 1:64, eight of 1:128 and two of 1:256. All cats were semi-domiciled, but lifestyle and diet were not assessed. T. gondii infection is rarely symptomatic and the severity depends on the pathophysiological state. Brazil has high prevalence rates and requires effective care in the management of cats. In Mineiros, 29 reports of infected pregnant women were identified. Although infection in humans occurs primarily through consumption of undercooked meat, ingestion of oocysts in feces of cats is a possibility. Therefore, state public health institutions and veterinarians should promote health education, highlighting food hygiene. Since, when symptomatic, the disease presents severe clinical conditions and, in some cases, irreversible.

A toxoplasmose é uma doença cosmopolita causada pelo Toxoplasma gondii, um protozoário obrigatório que afeta animais endotérmicos, incluindo o homem. Sendo uma zoonose que pode causar abortos, malformações fetais e cegueira. Os gatos são os hospedeiros definitivos, atuando como fonte de disseminação. Para coletar dados epidemiológicos sobre a distribuição do T. gondii em Mineiros, Goiás, amostras de gatos domésticos foram analisadas para verificar a circulação do protozoário e reforçar as medidas preventivas. Noventa e quatro amostras de gatos domésticos, independente de sexo, idade e raça, foram processadas aleatoriamente nos bairros de Mineiros, Goiás. Foram submetidos ao teste de anticorpo fluorescente indireto para pesquisa de anticorpos contra T. gondii, utilizando como antígenos taquizoítos e conjugado anti-cat IgG (1:16 foram positivos). Das 94 amostras analisadas, 35 (37,2%) eram reagentes, das quais quatro tiveram titulação de 1:16, dez de 1:32, onze de 1:64, oito de 1:128 e duas de 1:256. Todos os gatos eram semi-domiciliados, mas o estilo de vida e a dieta não foram avaliados. A infecção por T. gondii raramente é sintomática e a gravidade depende do estado fisiopatológico. O Brasil possui altas taxas de prevalência e requer cuidados efetivos no manejo dos gatos. Em Mineiros, foram identificadas 29 notificações de gestantes infectadas. Embora a infecção em humanos ocorra principalmente pelo consumo de carne mal cozida, a ingestão de oocistos nas fezes de gatos é uma possibilidade. Portanto, as instituições estaduais de saúde pública e os médicos veterinários devem promover a educação em saúde, destacando a higiene alimentar. Já que, quando sintomática, a doença apresenta quadros clínicos graves e, em alguns casos, irreversíveis.

La toxoplasmosis es una enfermedad cosmopolita causada por Toxoplasma gondii, un protozoario obligado que afecta a animales endotérmicos, incluido el hombre. Ser una zoonosis que puede provocar abortos, malformaciones fetales y ceguera. Los gatos son los huéspedes definitivos, actuando como fuente de diseminación. Con el fin de recolectar datos epidemiológicos sobre la distribución de T. gondii en Mineiros, Goiás, se analizaron muestras de gatos domésticos para verificar la circulación del protozoario y reforzar las medidas preventivas. Noventa y cuatro muestras de gatos domésticos, independientemente de género, edad y raza, fueron procesadas al azar en los barrios de Mineiros, Goiás. Se sometieron a la prueba de anticuerpos fluorescentes indirectos contra T. gondii, utilizando como antígenos taquizoítos e IgG anti-cat conjugada (1:16 fueron positivos). De las 94 muestras analizadas, 35 (37,2%) fueron reactivos, de los cuales cuatro tuvieron una titulación de 1:16, diez de 1:32, once de 1:64, ocho de 1:128 y dos de 1:256. Todos los gatos tenían domicilio parcial, pero no se evaluó el estilo de vida ni la dieta. La infección por T. gondii rara vez es sintomática y la gravedad depende del estado fisiopatológico. Brasil tiene altas tasas de prevalencia y requiere una atención eficaz en el manejo de los gatos. En Mineiros se identificaron 29 casos de mujeres embarazadas infectadas. Aunque la infección en los seres humanos se produce principalmente a través del consumo de carne poco cocida, la ingestión de ovoquistes en las heces de los gatos es una posibilidad. Por lo tanto, las instituciones estatales de salud pública y los veterinarios deben promover la educación sanitaria, destacando la higiene alimentaria. Dado que, cuando sintomática, la enfermedad presenta cuadros clínicos severos y, en algunos casos, irreversibles.

Seropositivity for Rickettsia spp. and Ehrlichia spp. in the human population of Mato Grosso, Central-Western Brazil.

INTRODUCTION: The epidemiology of Rickettsia and Ehrlichia species infection is underestimated in Mato Grosso State. METHODS:: Serum samples obtained during a Dengue outbreak in 2011-2012 were tested via indirect immunofluorescence and/or ELISA. RESULTS:: Samples from 19/506 (3.8%) patients presented antibodies for at least one of three Rickettsia species; 2/506 (0.4%) samples reacted against Ehrlichia canis. Most afflicted patients are residents of cities from the south-central region of the state, where these diseases have been reported in animals. CONCLUSIONS:: These results show serological evidence of human exposure to Rickettsia and Ehrlichia species in Mato Grosso State.

Epidemiological aspects of the occurrence of anti-Sarcocystis spp. andanti-Toxoplasma gondii antibodies in dogs in the Northern region of Brazil / Aspectos epidemiológicos da ocorrência de anticorpos anti-Sarcocystisspp. e anti-Toxoplasma gondii em cães da região norte do Brasil

We conducted aseroepidemiological study on the occurrence of anti-Sarcocystisspp. and anti-Toxoplasma gondii antibodies in dogs from family farming properties in the municipality of Ji-Paraná, Rondônia.Blood samples were collected from apparently healthy dogs between September 2012 and November 2013. In total, 181 blood serum samples were analyzed using an indirect immunofluorescence assay, among which 57 (31.49%) and 20 (11.04%) were positive for anti-T. gondii and anti-Sarcocystis spp., respectively. Statistical analyses showed that the type of food fed to the dogs was associated with the occurrence of anti-Sarcocystisspp. antibodies. In contrast, age and access to bovine carcasses were the risk factors for anti-T. gondii.The high occurrence of seropositive dogs for Sarcocystis spp. and T. gondii evidences the wide distribution of these agents in the studied area, possibly due to human and animal exposure to these protozoan species. In addition, anti-T. gondii antibodies were directly proportional to dog age. The increase in the number of positive animals with age was statistically significant. Furthermore, high antibody titers (up to 800) against Sarcocystis spp. in dogs suggest the possibility of recent exposure, in addition to environmental contamination by oocysts/sporocysts eliminated by the feces of these animals.

Conduzimos um estudo soroepidemiológico sobre a ocorrência de anticorpos anti- Sarcocystis spp. e anti-Toxoplasma gondiiem cães de propriedades de agricultura familiar no município de Ji-Paraná, Rondônia. Amostras de sangue foram coletadas de cães aparentemente saudáveis, entre setembro de 2012 e novembro de 2013. Ao todo, foram analisados 181 soros sanguíneos por meio do ensaio de imunofluorescência indireta, sendo positivas 57 (31,49%) e 20 (11,04%) amostras para anticorpos anti-T. gondii e anti-Sarcocystis spp., respectivamente. As análises estatísticas demonstraram que o tipo de alimentação fornecida aos cães esteve associado à ocorrência de anticorpos anti-Sarcocystis spp. Em contraste a idade e o acesso à carcaça bovina foram fatores de risco para a presença de anticorpos anti-T. gondii. A alta ocorrência de cães soropositivos para Sarcocystis spp. e T. gondii evidencia a ampla distribuição desses agentes na área estudada, possivelmente devido à exposição humana e animal a essas espécies de protozoários. Além disso, o resultado dos anticorpos anti-T. gondii relacionados a idade do cão mostraram diferença estatística, com aumento significativo no número de animais positivos com a idade. Além disso, altos títulos de anticorpos (até 800) contra Sarcocystis spp. em cães sugerem a possibilidade de exposição recente, além da contaminação ambiental por oocistos/esporocistos eliminados pelas fezes desses animais.

Ocorrência de Babesia sp, Ehrlichia canis e Hepatozoon canis em cães domiciliados, em dois municípios do estado do Espírito Santo - Brasil / Occurrence of Babesia sp, Ehrlichia canis and Hepatozoon canis in domiciled dogs in two municipalities of the state of Espírito Santo - Brazil / Ocurrencia de Babesia sp, Ehrlichia canis y Hepatozoon canis em perros domiciliados em dos municipios del estado de Espírito Santo - Brasil

As doenças transmitidas por carrapatos são afecções de grande importância na clínica médica de pequenos animais, devido à alta casuística e ampla distribuição vetorial no território brasileiro. Os principais agentes responsáveis pelas infecções em cães são Babesia sp., Ehrlichia canis e Hepatozoon canis. Os animais infectados são assintomáticos ou apresentam sinais clínicos inespecíficos, sendo necessário a utilização de testes diagnósticos para definição do agente etiológico, e diagnóstico seguro. O objetivo do presente estudo foi determinar a ocorrência desses micro-organismos em cães naturalmente infectados, domiciliados nos municípios de Vila Velha e Anchieta, Espírito Santo, utilizando diferentes testes de detecção: Reação em cadeia polimerase (PCR), sorologia para detecção de anticorpos anti Ehrlichia canis e pesquisa de hematozoários em esfregaço sanguíneo. Foram analisadas 65 amostras de sangue obtidas por venopunção de veia cefálica de cães. No teste de PCR, 4,62% dos animais foram positivos para Babesia vogeli e 1,54% para Ehrlichia canis sendo os resultados para Hepatozoon canis negativos. No teste sorológico para E. canis 90,77% dos animais foram positivos para a presença de anticorpos, e na pesquisa em lâminas de esfregaço sanguíneo 3,02% apresentavam outros hemoparasitas. Os resultados indicam a dispersão desses hemoparasitas na população canina da região de estudo, entretanto com baixa ocorrência. O teste de PCR demonstrou-se como o mais sensível no qual Babesia vogeli foi o agente mais observado.(AU)

Tick-borne diseases are diseases of great importance in the medical practice of small animals, due to the high casuistry and wide vectorial distribution in the Brazilian territory. The main agents responsible for infections in dogs are Babesia sp., Ehrlichia canis and Hepatozoon canis. Infected animals are asymptomatic or present nonspecific clinical signs, requiring the use of diagnostic tests to define the etiologic agent, and safe diagnosis. The objective of the present study was to determine the occurrence of these microorganisms in naturally infected dogs domiciled in the municipalities of Vila Velha and Anchieta, Espírito Santo, using different detection tests: polymerase chain reaction (PCR), serology to detect antibodies against Ehrlichia canis and research of hematozoa in blood smears. Sixty-five blood samples obtained by venipuncture of the cephalic vein of dogs were analyzed. In the PCR test, 4.62% of the animals were positive for Babesia vogeli and 1.54% for Ehrlichia canis, and the results for Hepatozoon canis were negative. In the serological test for E. canis, 90.77% of the animals were positive for the presence of antibodies, and in the research in blood smear slides, 3.02% presented other hemoparasites. The results indicate the dispersion of these hemoparasites in the canine population of the study region, however with low occurrence. The PCR test proved to be the most sensitive, in which Babesia vogeli was the most observed agent.(AU)

Las enfermedades transmitidas por garrapatas son enfermedades de gran importancia en la práctica médica de los pequeños animales, debido a la alta casuística y amplia distribución vectorial en el territorio brasileño. Los principales agentes responsables de las infecciones en los perros son Babesia sp., Ehrlichia canis y Hepatozoon canis. Los animales infectados son asintomáticos o presentan signos clínicos inespecíficos, siendo necesario el uso de pruebas diagnósticas para la definición del agente etiológico, y el diagnóstico seguro. El objetivo del presente estudio fue determinar la ocurrencia de estos microorganismos en perros infectados naturalmente, domiciliados en los municipios de Vila Velha y Anchieta, Espírito Santo, utilizando diferentes pruebas de detección: reacción en cadena de la polimerasa (PCR), serología para detectar anticuerpos anti Ehrlichia canis e investigación de hematozoos en frotis de sangre. Se analizaron sesenta y cinco muestras de sangre obtenidas por venopunción de la vena cefálica de los perros. En la prueba PCR, el 4,62% de los animales fueron positivos para Babesia vogeli y el 1,54% para Ehrlichia canis, y los resultados para Hepatozoon canis fueron negativos. En la prueba serológica para E. canis, el 90,77% de los animales fueron positivos a la presencia de anticuerpos, y en la investigación en láminas de frotis de sangre el 3,02% presentaron otros hemoparásitos. Los resultados indican la dispersión de estos hemoparásitos en la población canina de la región de estudio, aunque con una baja presencia. La prueba PCR resultó ser la más sensible, en la que Babesia vogeli fue el agente más observado.(AU)

Canine gastrointestinal pythiosis: a case report in Brazilian Pantanal from a diagnostic approach / Pitiose gastrointestinal canina: relato de caso no Pantanal brasileiro a partir de uma abordagem diagnóstica

Pythiosis is caused by an aquatic fungus-like organism (Pythium insidiosum). It is considered an important public health issue as it can affect both animals and humans. This paper reports a case of gastrointestinal pythiosis in a dog. The patient was hospitalized for four days, during which the animal received supportive and symptomatic treatment. But the applied treatment was unsuccessful and the dog's clinical condition worsened, culminating in death. Complementary imaging tests such as radiography and ultrasonography, as well as hematological tests, were performed during the hospitalization period. The definitive diagnosis was reached in the postmortem as macroscopic and microscopic characteristics suggested the presence of intestinal granuloma and accentuated multifocal suppurative necrotic enteritis. Additionally, the histological evaluation revealed morphological structures compatible with P. insidiosum. Also, the results of nested PCR performed showed partial amplification (105 bp) of the ITS1 region of the ribosomal gene of P. insidiosum.(AU)

A pitiose é causada por um organismo aquático semelhante a um fungo (Pythium insidiosum) e considerada um importante problema de saúde pública, pois pode afetar animais e humanos. Este artigo relata um caso de pitiose gastrointestinal em um cão. O paciente ficou internado por quatro dias, período em que o animal recebeu tratamento de suporte e sintomático. No entanto, o tratamento aplicado não teve sucesso e o quadro clínico do cão piorou, culminando com a morte. Exames de imagem complementares, como radiografia e ultrassonografia, bem como exames hematológicos, foram realizados durante o período de internação. O diagnóstico definitivo foi feito na autópsia, pois as características macroscópicas e microscópicas sugeriam a presença de granuloma intestinal e acentuada enterite necrótica multifocal supurativa. Além disso, a avaliação histológica revelou estruturas morfológicas compatíveis com P. insidiosum. Além disso, a nested PCR foi realizada e mostrou amplificação parcial (105 pb) da região ITS1 do gene ribossomal de P. insidiosum.(AU)

Seroprevalence and hematological abnormalities associated with Ehrlichia canis in dogs referred to a veterinary teaching hospital in central-western Brazil / Soroprevalência e alterações hematológicas associadas com Ehrlichia canis em cães encaminhados a um hospital veterinário-escola no centro-oeste do Brasil

Ticks are significant parasites of dogs in the tropics, where tick-borne pathogens are highly prevalent, especially in areas where tick control measures are frequently neglected. This study investigated the seroprevalence and hematological abnormalities associated with Ehrlichia canis in dogs referred to a veterinary teaching hospital in Central-western Brazil. Out of 264 dogs tested for anti-Ehrlichia canis antibodies by an indirect immunofluorescence assay (IFA), 59.1% (156/264) were positive. Seropositivity was significantly associated to anemia and thrombocytopenia, alone or in combination, and to leukopenia. Conversely, there were no differences in terms of seroprevalence according to sex, breed and age. This study demonstrated that dogs referred to a veterinary teaching hospital in Central-western Brazil are highly exposed to E. canis and that seropositive dogs are more likely to present hematological abnormalities, particularly anemia, thrombocytopenia and leukopenia. To our knowledge, this is the first study on detection of anti-E. canis antibodies by means of IFA among dogs in the state of Goiás. These findings highlighted the need for increasing awareness among dog owners regarding tick control measures in Central-western Brazil, ultimately to reduce the risk of exposure to E. canis and other tick-borne pathogens.

Carrapatos são importantes parasitos de cães nos trópicos, onde patógenos transmitidos por carrapatos são altamente prevalentes, especialmente em áreas onde as medidas de controle de carrapatos são frequentemente negligenciadas. O estudo investigou a soroprevalência e as anormalidades hematológicas associadas à Ehrlichia canis em cães encaminhados para um hospital veterinário-escola no Centro-oeste do Brasil. Dos 264 cães testados para anticorpos anti-Ehrlichia canis por meio da reação de imunofluorescência indireta (RIFI), 59.1% (156/264) foram positivos. A soropositividade foi associada significativamente à anemia e trombocitopenia, isoladamente ou em combinação, e à leucopenia. Por outro lado, não houve diferenças quanto à soroprevalência segundo sexo, raça e idade. Este estudo demonstrou que cães encaminhados a um hospital veterinário-escola na região Centro-oeste do Brasil são altamente expostos à E. canis, e que cães soropositivos têm maior probabilidade de apresentar alterações hematológicas, principalmente anemia, trombocitopenia e leucopenia. Para o nosso conhecimento, este é o primeiro estudo sobre a detecção de anticorpos anti-E. canis por meio da RIFI em cães do estado de Goiás. Essas descobertas destacam a necessidade de aumentar a conscientização entre os proprietários de cães em relação às medidas de controle do carrapato no Centro-oeste do Brasil, em última análise, para reduzir o risco de exposição ao E. canis e outros patógenos transmitidos por carrapatos.

Serological evidence of exposure to tick-borne agents in opossums (Didelphis spp.) in the state of São Paulo, Brazil.

This work involved a serological investigation of tick-borne pathogens in opossums in eight municipalities of the state of São Paulo, Brazil. Serum samples from 109 opossums (91 Didelphis aurita and 18 Didelphis albiventris) were tested to detect antibodies to Rickettsia rickettsii (Taiaçu strain, 1:64 cut-off) and Ehrlichia canis (São Paulo strain, 1:40 cut-off), by indirect immunofluorescence assay (IFA); and against Borrelia burgdorferi (strain G39/40) by enzyme-linked immunosorbent assay (ELISA). The presence of antibodies to anti-R. rickettsii, anti-E. canis and anti-B. burgdorferi was detected in 32 (29.35%), 16 (14.67%) and 30 (27.52%) opossums, respectively. Opossum endpoint titers ranged from 64 to 1,024 for R. rickettsii, from 40 to 160 for E. canis, and from 400 to >51,200 for B. burgdorferi. These serological results suggest that opossums have been exposed to Rickettsia spp., Ehrlichia spp., and B. burgdorferi-related agents in the state of São Paulo. Our study underscores the need for further research about these agents in this study area, in view of the occurrence of Spotted Fever and Baggio-Yoshinari Syndrome disease in humans in the state of São Paulo, Brazil.

Anti-Ehrlichia properties of the dichloromethane fraction of Ageratum conyzoides associated with doxycycline: In vitro study / Propriedades anti-Ehrlichia da fração diclorometânica de Ageratum conyzoides associada com doxiciclina: estudo in vitro

ABSTRACT: The increasing number of cases of canine ehrlichiosis caused by Ehrlichia canis in hospitals and veterinary clinics has demonstrated the need for a new drug protocol for this disease. Doxycycline is used to treat ehrlichiosis, but the resistance of the microorganism to this treatment protocol, as well as the various side effects to the animals, has become a concern. Several studies have shown a positive interaction with extracts of plants and drugs, which allow for the reduction of the concentration necessary to produce the desired effect, minimizing adverse effects. This study determined the efficiency of the combination of the dichloromethane (DCM) fraction of Ageratum conyzoides L. with anti-Ehrlichia activity and doxycycline by using the checkerboard assay. Plant material was collected in São Luís, northeastern Brazil, followed by extraction in MeOH: H2O (8:2) and partitioning of the DCM fraction. After determining the Minimum Inhibitory Concentration (MIC) of the fraction under study against DH82 cells infected with Ehrlichia canis, it was combined with doxycycline to derive the Fractional Inhibitory Concentration Index (CIF Index). A reduction of 5.83 times the doxycycline minimum inhibitory concentration was observed, showing that this fraction of A. conyzoides composed predominantly by the class of lignans, identified by mass spectrometry notably intensified the activity of doxycycline against E. canis, resulting in a synergistic effect.

RESUMO: O crescente número de casos de erliquiose canina por Ehrlichia canis em hospitais e clínicas veterinárias tem demonstrado a necessidade de um novo protocolo de medicamentos para essa doença. A doxiciclina é usada para tratar a erliquiose, mas a resistência do microrganismo a esse protocolo de tratamento, bem como os diversos efeitos colaterais para os animais, tornou-se uma preocupação. Vários estudos têm demonstrado interação positiva com extratos de plantas e fármacos, que permitem a redução da concentração necessária para produzir o efeito desejado, minimizando os efeitos adversos. Este estudo determinou a eficiência da combinação da fração diclorometânica (DCM) de Ageratum conyzoides L. com atividade anti-Ehrlichia canis associada com doxiciclina por meio do ensaio de Checkerboard. O material vegetal foi coletado em São Luís, Maranhão, nordeste do Brasil, seguido pela extração em MeOH:H2O (8:2) e partição da fração diclorometânica. Após a determinação da Concentração Inibitória Mínima (CIM) da fração em estudo frente às células DH82 infectadas com Ehrlichia canis, a mesma foi combinada com a doxiciclina para derivação do Índice de Concentração Inibitória da Fração (Índice CIF). Observou-se uma redução de 5,83 vezes a concentração inibitória mínima da doxiciclina mostrando que esta fração de A. conyzoides, composta predominantemente por lignanas identificadas por espectrometria de massas, notavelmente intensificou a atividade desse fármaco contra E. canis, resultando em um efeito sinérgico.

Detection of Leptospira spp. and Brucella abortus antibodies in free-living jaguars (Panthera onca) in two protected areas of northern Pantanal, Brazil.

This study aimed to assess the exposure of free-living jaguars (Panthera onca) to Leptospira spp. and Brucella abortus in two conservation units in the Pantanal of Mato Grosso, Brazil. The presence of antibodies in blood samples of eleven jaguars was investigated using autochthonous antigens isolated in Brazil added to reference antigen collection applied to diagnosis of leptospirosis by Microscopic Agglutination Test (MAT). The Rose Bengal test was applied for B. abortus antibodies. Two (18.2%) jaguars were seroreactive for the Leptospira spp. antigen and the serovar considered as most infective in both animals was a Brazilian isolate of serovar Canicola (L01). All jaguars were seronegative for B. abortus. These data indicate that the inclusion of autochthonous antigens in serological studies can significantly increase the number of reactive animals, as well as modify the epidemiological profile of Leptospira spp. infection.

Prevalence of Neospora caninum and Toxoplasma gondii antibodies in dogs in the municipality of Poconé, state of Mato Grosso, Brazil / Prevalência de anticorpos contra Neospora caninum e Toxoplasma gondii no município de Poconé, estado de Mato Grosso, Brasil

This study evaluated the presence of Neospora caninum and Toxoplasma gondii antibodies in 320 domestic dogs living in urban and rural areas of the municipality of Poconé, located in the Pantanal wetlands of the state of Mato Grosso, Brazil. Anti- N. caninum (cut-off point 50) and anti- T. gondii (cut-off point 16) antibodies were detected by means of immunofluorescence assays (IFA), using NC-Bahia and RH strains as antigens, respectively. Anti- N. caninum antibodies were detected in 69 (21.56%; 95% CI: 17.27%-26.56%) dogs, 31 (44.93%) of which lived in urban areas and 38 (55.07%) in rural areas, and endpoint titers ranged from 50 to 3200. Anti- T. gondii antibodies were found in 132 (41.25%; 95% CI: 35.84%-46.87%) dogs, 58 (43.94%) from urban areas and 74 (56.06%) from rural areas, and endpoint titers ranged from 16 to 8192. A total of 33 dogs (10.3%) (12 urban and 21 rural animals) reacted to both agents (P <0.05). This study showed a higher prevalence of anti- T. gondii compared to anti- N. caninum antibodies in the dogs living in the Pantanal region, suggesting that both protozoans circulate in the studied region. This information is relevant, in view of its implications for animal and public health.

Este estudo avaliou a presença de anticorpos contra N. caninum e T. gondii em 320 cães domésticos das áreas urbana e rural do município de Poconé, no estado do Mato Grosso, região pantaneira do Pantanal. A Reação de Imunofluorescência Indireta (RIFI) foi utilizada para detectar anticorpos anti- N. caninum (ponto de corte igual a 50) e anticorpos anti- T. gondii (ponto de corte igual a 16) utilizando as cepas NC-Bahia e RH como antígenos, respectivamente. Anticorpos anti- N. caninum foram detectados em 69 (21,56%; IC 95%: 17,27%-26,56%) cães, dos quais 31 (44,93%) eram da área urbana e 38 (55,07%) eram da área rural e os títulos variaram entre 50 a 3200. Em relação aos anticorpos contra T. gondii 132 (41,25%; IC 95%: 35,84%-46,87%) cães foram sororeagentes sendo que 58 (43,94%) eram da área urbana e 74 (56,06%) da área rural e os títulos variaram entre 16 a 8192. Um total de 33 (10,31%) (12 cães urbanos e 21 cães rurais) cães reagiram para ambos os agentes testados pela RIFI (P <0,05). Este estudo mostrou uma maior ocorrência de anticorpos contra T. gondii em relação a N. caninum nos cães avaliados, mostrando que na região do Pantanal há circulação desses importantes protozoários. Essa informação é relevante dada sua importância para a saúde animal e pública

Antibodies anti-Toxoplasma gondii and anti-Neospora caninum in backyard pigs from the state of Mato Grosso, Brazil / Anticorpos anti-Toxoplasma gondii e anti-Neospora caninum em porcos de criações de fundo de quintal do Estado de Mato Grosso, Brasil

Abstract To estimate the seroprevalence of Toxoplasma gondii and Neospora caninum, using an indirect immunofluorescent assay (IFA), and identify the risk factors associated, serum samples were collected from 1,070 pigs from 320 backyard pig farming in the of Mato Grosso state. The animal-level seroprevalence of T. gondii and N. caninum was 32.48% and 13.49%, respectively, with a herd seroprevalence of 55.63% for T. gondii and 27.81% for N. caninum. Feeding the animals with leftovers increases the probability of the presence of anti-T. gondii antibodies in pigs by 1.09-fold. Unlike to T. gondii, feeding with leftovers was found to be negatively associated with N. caninum seropositivity in farm-level analysis and in the animal-level model, so decreasing the chances of positivity. Yet, age was considered a risk factor for N. caninum seropositivity. Further studies are necessary to evaluate the impact of T. gondii infection on backyard pig farming production, and its importance as a source of toxoplasmosis infection in humans in the Mato Grosso state, as well as, the role of domestic pigs in the epidemiology of neosporosis.

Resumo Para estimar a soroprevalência de Toxoplasma gondii e Neospora caninum, utilizando a reação de imunofluorescência indireta (RIFI), e identificar os fatores de risco associados, foram coletadas amostras de soro de 1.070 suínos provenientes de 320 criatórios de fundo de quintal no Estado de Mato Grosso. A soroprevalência para nível animal de T. gondii e N. caninum foi de 32,48% e 13,49%, respectivamente, com uma soroprevalência de rebanho de 55,63% para T. gondii e 27,81% para N. caninum. Alimentar os animais com sobras aumenta a probabilidade da presença de anticorpos anti-T. gondii em porcos em 1,09 vezes. Ao contrário de T. gondii, a alimentação baseada em sobras foi encontrada como negativamente associada à soropositividade para N. caninum na análise em nível de fazenda e no modelo em nível animal, diminuindo assim as chances de positividade. Ainda, a idade foi considerada um fator de risco para a soropositividade para N. caninum. Mais estudos são necessários para avaliar o impacto da infecção por T. gondii na produção de suinocultura de quintal e sua importância como fonte de infecção para toxoplasmose em humanos no Estado de Mato Grosso, bem como o papel dos suínos domésticos na epidemiologia da neosporose.

Soropositividade para Leptospira sp. e sorogrupos predominantes em cães do Pantanal Brasileiro / Seropositivity for Leptospira sp. and predominant serogroups in dogs from the Brazilian Pantanal

Em face à grande importância que a leptospirose possui no contexto sanitário mundial, tanto no aspecto humano como animal, este estudo teve por objetivo realizar a pesquisa de anticorpos anti-Leptospira sp. pela técnica de Soroaglutinação Microscópica (SAM) em 429 amostras de soros de cães provenientes de quatro municípios (Poconé/MT, Santo Antônio de Leverger/MT, Barão de Melgaço/MT e Corumbá/MS) localizados na região do Pantanal Brasileiro, bem como foram verificadas possíveis associações entre os resultados dos exames sorológicos e respostas aos questionários epidemiológicos aplicados aos proprietários. Do total de cães avaliados pela SAM (título ≥100), verificou-se que 34 (7,93%; IC 95%: 5,63%-11,00%) cães tinham anticorpos anti-Leptospira sp. Os títulos encontrados variaram entre 100 e 1600 e todos os municípios analisados tinham cães sororreagentes ao agente pesquisado. O sorogrupo reator mais frequente foi o Icterohaemorrhagiae, seguido pelo Australis. Por outro lado, foram observadas menores proporções de cães reagentes aos sorogrupos Tarassovi, Hebdomadis, Autumnalis e Grippotyphosa. As variáveis associadas com a ocorrência de leptospirose foram habitat rural (P<0,01) e área alagável (P=0,01). Estes resultados demonstram que os cães da região pantaneira tiveram contato com agentes do gênero Leptospira, o que representa uma informação relevante para a saúde pública local devido à importância zoonótica da doença.

Given the great importance that leptospirosis has the global health context, both in human and animal aspect, this study aimed to search for antibodies anti-Leptospira sp. by the technique of microscopic agglutination test (MAT) in 429 samples of sera from dogs from four municipalities (Poconé/MT, Santo Antônio de Leverger/MT, Barão de Melgaço/MT and Corumbá/MS) located in the Brazilian Pantanal region, in order for determine associations between the results of the serological tests and answers to epidemiological questionnaires applied to owners. Of the total dogs evaluated by MAT (titer ≥100), it was verified that 34 (7.93%, 95% CI: 5.63% -11.00%) dogs had antibodies against Leptospira sp. The titers found ranged from 100 to 1600 and all municipalities analyzed had seroreactive dogs for the investigated agent. The most frequent serogroup reactor was Icterohaemorrhagiae, followed by Australis. On the other hand, smaller proportions of reactive dogs were observed for serogroups Tarassovi, Hebdomadis, Autumnalis and Grippotyphosa. The variables associated with the occurrence of leptospirosis were rural habitat (P<0.01) and flooded area (P=0.01). These results demonstrate that dogs from the Pantanal region had contact with agents of the genus Leptospira, which represents information relevant to local public health due to the zoonotic importance of the disease.

Soropositividade para Leptospira sp. e sorogrupos predominantes em cães do Pantanal Brasileiro / Seropositivity for Leptospira sp. and predominant serogroups in dogs from the Brazilian Pantanal

Em face à grande importância que a leptospirose possui no contexto sanitário mundial, tanto no aspecto humano como animal, este estudo teve por objetivo realizar a pesquisa de anticorpos anti-Leptospira sp. pela técnica de Soroaglutinação Microscópica (SAM) em 429 amostras de soros de cães provenientes de quatro municípios (Poconé/MT, Santo Antônio de Leverger/MT, Barão de Melgaço/MT e Corumbá/MS) localizados na região do Pantanal Brasileiro, bem como foram verificadas possíveis associações entre os resultados dos exames sorológicos e respostas aos questionários epidemiológicos aplicados aos proprietários. Do total de cães avaliados pela SAM (título 100), verificou-se que 34 (7,93%; IC 95%: 5,63%-11,00%) cães tinham anticorpos antiLeptospira sp. Os títulos encontrados variaram entre 100 e 1600 e todos os municípios analisados tinham cães sororreagentes ao agente pesquisado. O sorogrupo reator mais frequente foi o Icterohaemorrhagiae, seguido pelo Australis. Por outro lado, foram observadas menores proporções de cães reagentes aos sorogrupos Tarassovi, Hebdomadis, Autumnalis e Grippotyphosa. As variáveis associadas com a ocorrência de leptospirose foram habitat rural (P 0,01) e área alagável (P=0,01).

Given the great importance that leptospirosis has the global health context, both in human and animal aspect, this study aimed tosearch for antibodies anti-Leptospira sp. by the technique of microscopic agglutination test (MAT) in 429 samples of sera from dogsfrom four municipalities (Poconé/MT, Santo Antônio de Leverger/MT, Barão de Melgaço/MT and Corumbá/MS) located in the BrazilianPantanal region, in order for determine associations between the results of the serological tests and answers to epidemiologicalquestionnaires applied to owners. Of the total dogs evaluated by MAT (titer ≥100), it was verified that 34 (7.93%, 95% CI: 5.63%-11.00%) dogs had antibodies against Leptospira sp. The titers found ranged from 100 to 1600 and all municipalities analyzed hadseroreactive dogs for the investigated agent. The most frequent serogroup reactor was Icterohaemorrhagiae, followed by Australis.On the other hand, smaller proportions of reactive dogs were observed for serogroups Tarassovi, Hebdomadis, Autumnalis andGrippotyphosa. The variables associated with the occurrence of leptospirosis were rural habitat (P<0.01) and flooded area (P=0.01).These results demonstrate that dogs from the Pantanal region had contact with agents of the genus Leptospira, which representsinformation relevant to local public health due to the zoonotic importance of the disease.