Serological Evidence Supporting the Occurrence of Ehrlichia chaffeensis or a Closely Related Species in Brazilian Dogs.

Ehrlichia chaffeensis is a tick-borne bacterium that causes human monocytotropic ehrlichiosis, an emerging life-threatening disease in humans transmitted by Amblyomma americanum. Although most studies have reported bacterial isolations and clinical cases in the United States, their occurrence is not restricted to North America. Some studies in the Southern Cone of South America have molecularly detected a close phylogenetic relative of E. chaffeensis in ticks and wild mammals. Even so, many gaps must be filled to confirm the presence of this agent in the region. To add new data on this issue, we report the first detection of specific anti-E. chaffeensis antibodies in dogs collected from all regions of Brazil. By means of IFA and ELISA with crude and specific antigens of E. chaffeensis, sera from 1134 dogs were analyzed. Serological analyses using ELISA showed nine (0.7%) seropositive dogs, with seven of them exhibiting IFA titers ranging from 160 to 5120. All regions of Brazil had at least one seropositive dog. Our results support the evidence for the occurrence of E. chaffeensis in South America. As dogs have a close relationship with humans, they can be used as an environmental sentinel for these infections because they can act as a bridge to human parasitism or infection with ehrlichial agents.

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.

Novel Ehrlichia canis genogroup in dogs with canine ehrlichiosis in Cuba.

BACKGROUND: Canine monocytic ehrlichiosis (CME) is caused by the tick-borne pathogen Ehrlichia canis, an obligate intracellular Gram-negative bacterium of the family Anaplasmataceae with tropism for canine monocytes and macrophages. The trp36 gene, which encodes for the major immunoreactive protein TRP36 in E. canis, has been successfully used to characterize the genetic diversity of this pathogen in different regions of the world. Based on trp36 sequence analysis, four E. canis genogroups, United States (US), Taiwan (TWN), Brazil (BR) and Costa Rica (CR), have been identified. The aim of this study was to characterize the genetic diversity of E. canis in Cuba based on the trp36 gene. METHODS: Whole blood samples (n = 8) were collected from dogs found to be infested with the tick vector Rhipicephalus sanguineus sensu lato (s.l.) and/or presenting clinical signs and symptoms of CME. Total DNA was extracted from the blood samples and trp36 fragments were amplified by PCR. Nucleotide and protein sequences were compared using alignments and phylogenetic analysis. RESULTS: Four of the trp36 sequences obtained (n = 8) fall within the phylogenetic cluster grouping the US genogroup E. canis strains. The other E. canis trp36 sequences formed a separate and well-supported clade (94% bootstrap value) that is phylogenetically distant from the other major groups and thus represents a new genogroup, herein designated as the 'Cuba (CUB) genogroup'. Notably, dogs infected with the CUB genogroup presented frequent hemorrhagic lesions. CONCLUSIONS: The results of this study suggest that genetic diversification of E. canis in Cuba is associated with the emergence of E. canis strains with increased virulence.

Concentrations of tissue inhibitor of matrix metalloproteinase-1 and hyaluronic acid in canine amniotic membranes cryopreserved for different time points and its effects in dogs with complicated corneal ulcers.

OBJECTIVES: To determine the concentrations of total protein (TP), tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) and hyaluronic acid (HA) in amniotic membranes (AMs) harvested from placentas of bitches of different ages and cryopreserved for different time points. The outcomes of complicated corneal defects of dogs repaired with AMs stored for the same time points were also evaluated. PROCEDURES: Ten cryopreserved canine AMs were stored for short term (2-50 days), middle term (92-210 days), or long term (256-357 days). TP was quantified by Bradford's test, whereas TIMP-1 and HA were quantified by ELISA. Twenty-one dogs that had an AM transplantation to restore deep or perforating corneal wounds were selected. RESULTS: TIMP-1 levels were lower in AMs cryopreserved for middle term (p = .02) and long term (p = .0009), when compared to AMs stored for short term. TP (p = .39) and HA (p = .18) concentrations in AMs did not differ among the storage time. TIMP-1 concentration in AMs correlated with storage time (R = -.65, p < .0001), while TP (R = -.33, p = .07) and HA concentrations did not (R = -.15, p = .41). The age of donors did not correlate with the components evaluated in the AMs. Corneal defects repaired with AMs stored for short term healed sooner than the ones repaired with AMs stored for middle (p < .01) and long term (p = .02). Additionally, TIMP-1 levels in AMs correlated negatively with the epithelization time (R = -.62, p = .002). Graft opacity was severe in 55% of cases. However, the HA levels in AMs correlated negatively with the opacification score (R = -.47, p = .03). Vision was observed in more patients who presented deep ulcers and descemetoceles, than in the ones with perforations (p = .004). CONCLUSIONS: TIMP-1 concentration in canine AMs significantly decreased over a year storage time, while TP and HA concentrations did not change during the same period. The age of donors did not correlate with the components evaluated in the AMs. Complicated corneal defects repaired with AMs cryopreserved for short term healed sooner and tended to be less opaque; however, satisfactory to optimal outcomes were achieved even in the eyes repaired with AMs stored for up to a year.

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.

Exposure of Domestic Cats to Distinct Ehrlichia canis TRP Genotypes.

Cats naturally exposed to Ehrlichia canis have been described in different regions of the world, but little is known about the genotypes associated with infection in these animals. To detect E. canis-specific antibodies and investigate the E. canis TRP genotypes in cats, serum samples from 76 domestic cats reactive to crude E. canis antigens by the indirect fluorescence antibody test (IFAT) were analyzed by ELISA, using E. canis-specific peptides (i.e., TRP19 and TRP36 /BR/US/CR). Of these, 25 (32.9%) cats reacted to at least one TRP peptide, confirming their specific exposure to E. canis. Eighteen (23.7%) cats reacted to TRP19, 15 (19.8%) to BRTRP36, and 11 (14.5%) to USTRP36, but none of them reacted to CRTRP36. Eight (10.5%) cats reacted to TRP19 but not to any TRP36 genotype, demonstrating the possible existence of a new E. canis genotype infecting felines. Nevertheless, this study provides the first report of anti-E. canis-specific antibodies in domestic cats.

Geographic Distribution of Ehrlichia canis TRP Genotypes in Brazil.

Tandem repeat proteins (TRPs) are major immunoreactive proteins of Ehrlichia canis, which have been used in the serological diagnosis of different genotypes of the microorganism. TRP19 is preserved among different E. canis isolates expressed on both reticulate and dense-core cells and observed in the extracellular matrix or associated with the morula membrane. TRP36 is differentially expressed only on the surface of the dense-core form of the bacterium and exhibits more divergence among isolates. The aim of this study was to evaluate the distribution of the American (USTRP36), Brazilian (BrTRP36) and Costa Rican (CRTRP36) genotypes of E. canis in Brazil, using ELISA assays. Serum samples of 814 dogs from 49 municipalities from all over Brazil were analyzed. Our results showed that 33.9% of the samples were reactive to the USTRP36 genotype and 32.6% to the BrTRP36 genotype. The two genotypes appeared to occur equally throughout Brazil, although the frequency of seropositivity was lower in the south than in the country's other regions. Dogs that reacted to at least one of the synthetic peptides (TRP19 and TRP36) were 456 (56%). A few dogs (n = 5; 0.6%) reactive to the E. canis TRP36 genotype (CRTRP36) were also detected in the northeast and southern regions. We concluded that the American and Brazilian genotypes of E. canis are distributed evenly in Brazil, especially in the tropical region, while the temperate region in the south presented the lowest prevalence rates. This study offers the first report of dogs seropositive for the Costa Rican genotype in Brazil.

Uncommon Ehrlichia canis infection associated with morulae in neutrophils from naturally infected dogs in Brazil.

Ehrlichia and Anaplasma species are the most common tick-borne disease (TBD) pathogens in dogs worldwide. Ehrlichia canis, the aetiological agent of the Canine Monocytic Ehrlichiosis (CME), is known to replicate within the cytoplasm of mononuclear cells into clusters of organisms called morulae. However, detection of morulae in neutrophils is commonly observed in dogs infected by Ehrlichia ewingii or Anaplasma phagocytophilum. We report uncommon clinical cases of canine ehrlichiosis presenting morulae compatible with E. ewingii and A. phagocytophilum in dogs from two distinct regions of Brazil. Eight dogs were admitted to two veterinary teaching hospitals from Brazil, showing clinical or haematological signs suggestive of TBD. Blood or peritoneal fluid was withdrawn for haematological and cytologic analysis. All samples were evaluated by PCR assays for Ehrlichia and Anaplasma using genus-specific primers for dsb, 16S rRNA and groEL genes, followed by sequencing. Samples were also evaluated by nested PCR assays for the 16S rRNA gene of E. ewingii and groEL gene of A. phagocytophilum and Anaplasma platys. Seven dogs revealed thrombocytopenia, six dogs had monocytosis and five presented lymphopenia and anaemia. All dogs showed morulae structures compatible with Ehrlichia spp. in neutrophils and were PCR-positive for the dsb and 16S rRNA gene fragments of Ehrlichia, with sequences showing 100% identity with multiple E. canis sequences deposited in the GenBank™. Sequencing of 16S rRNA and groEL gene fragments from one PCR-positive dog showed 100% identity with A. platys. Overall, our data suggest that in endemic regions for E. canis, that is Brazil, the presence of morulae in neutrophils may indicate infection by this bacterium. Herein, morulae were also found in neutrophils present in the peritoneal fluid of a dog. Also, this is the first report of E. canis and Hepatozoon canis co-infection in neutrophils from naturally infected dogs confirmed by DNA sequencing.

Detection molecular of Rangelia vitalii in dogs from Parana State, Southern Brazil / Detecção molecular de Rangelia vitalii em cães do Estado do Paraná, Sul do Brasil

Abstract 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.

Resumo Rangelia vitalii infecta eritrócitos, leucócitos e células endoteliais de cães. O presente estudo objetivou relatar a detecção molecular confirmada por sequenciamento de R. vitalii no estado do Paraná e descrever as alterações clínicas, hematológicas e bioquímicas dos cães infectados. Três cães doentes da região metropolitana de Curitiba, PR, Brasil, foram submetidos a exame físico e exames laboratoriais que incluíram hematologia, bioquímica, reação em cadeia da polimerase (PCR) e sequenciamento genético. Os sinais clínicos incluíram apatia, anorexia e hemorragia. Piroplasmas intra-eritrocíticos e extracelulares foram encontrados em esfregaços de sangue periférico dos três cães. As amostras de sangue destes animais foram positivas para Babesia sp. pela PCR baseada no gene 18S rRNA. Os produtos de PCR dos três cães foram sequenciados e a análise de BLAST mostrou que as seqüências geradas por PCR eram altamente homólogas com as de R. vitalii previamente relatadas. Os achados hematológicos incluíram anemia grave, desvio de neutrófilos à esquerda regenerativo e trombocitopenia. Os níveis de uréia no soro aumentaram nos três cães, e os níveis de bilirrubina direta foram elevados em um cão.

Isolation and genome characterization of canine parvovirus type 2c in Brazil.

This study focused on the isolation and characterization of parvovirus in an infected dog in midwestern Brazil. Non-enveloped icosahedral parvovirus-like particles were isolated in CRFK cells and were allocated to a clade comprised of strains of CPV-2c, based on genome analysis. This is the first isolate of CPV-2c genomically characterized in Brazil.

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.

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.

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

Abstract 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.

Ehrlichia canis DNA in domestic cats parasitized by Rhipicephalus sanguineus sensu lato (s. l. ) ticks in Brazil ­ case report / Felinos domésticos parasitados por carrapato Rhipicephalus sanguineus sensu lato (s. l. ) infectados por Ehrlichia canis no Brasil ­ relato de caso

Ectoparasites can transmit pathogens, including bacteria such as Ehrlichia sp., which trigger infectious diseases in domestic animals. Little is known about the epidemiology of feline ehrlichiosis, although several studies have focused on elucidating the pathogenesis and transmission of this disease. This paper presents the first mutual infection by Ehrlichia sp. between a domestic cat and a Rhipicephalus sanguineus sensu lato (s.l.) tick removed from the animal. The cat and tick were tested by Polymerase Chain Reaction (PCR) to detect the dsb gene, and the analyzed sequences revealed samples 100% identical to E. canis. Based on this report, we discussed the importance of cats as E. canis reservoirs s and their position in the cycle of transmission between dogs and cats in Brazil.(AU)

Os ectoparasitos são capazes de transmitir patógenos incluindo algumas bactérias, como a Ehrlichia sp., causando doenças em animais domésticos. Pouco se conhece sobre a epidemiologia da erliquiose felina, embora alguns estudos já tenham sido realizados para elucidar a sua patogenia e transmissão. Este trabalho relata a primeira infecção mútua por Ehrlichia sp. entre um felino doméstico e o carrapato Rhipicephalus sanguineus sensu lato (s.l.) removido deste animal. Ambos foram testados pela Reação em Cadeia pela Polimerase (PCR) para detectar o gene dsb, e as sequências analisadas confirmaram amostras 100% idênticas à E. canis. Baseado neste relato é discutida a importância dos gatos como vetores de E. canis e sua posição no ciclo de transmissão de carrapatos entre cães e gatos no Brasil.(AU)

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

Abstract 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.

Resumo Rangelia vitalii infecta eritrócitos, leucócitos e células endoteliais de cães. O presente estudo objetivou relatar a detecção molecular confirmada por sequenciamento de R. vitalii no estado do Paraná e descrever as alterações clínicas, hematológicas e bioquímicas dos cães infectados. Três cães doentes da região metropolitana de Curitiba, PR, Brasil, foram submetidos a exame físico e exames laboratoriais que incluíram hematologia, bioquímica, reação em cadeia da polimerase (PCR) e sequenciamento genético. Os sinais clínicos incluíram apatia, anorexia e hemorragia. Piroplasmas intra-eritrocíticos e extracelulares foram encontrados em esfregaços de sangue periférico dos três cães. As amostras de sangue destes animais foram positivas para Babesia sp. pela PCR baseada no gene 18S rRNA. Os produtos de PCR dos três cães foram sequenciados e a análise de BLAST mostrou que as seqüências geradas por PCR eram altamente homólogas com as de R. vitalii previamente relatadas. Os achados hematológicos incluíram anemia grave, desvio de neutrófilos à esquerda regenerativo e trombocitopenia. Os níveis de uréia no soro aumentaram nos três cães, e os níveis de bilirrubina direta foram elevados em um cão.

Molecular assessment of the transplacental transmission of Toxoplasma gondii, Neospora caninum, Brucella canis and Ehrlichia canis in dogs.

Given the fact that numerous microbial species can be detected in pregnant female dogs, the objective of this study was to assess the transplacental transmission of Brucella canis, Ehrlichia canis, Neospora caninum and Toxoplasma gondii in stillborn puppies. This study involved 41 stillborn puppies, 78.6% of which were positive for T. gondii, 52.4% for N. caninum and 59.5% for B. canis. E. canis was not detected in any of the analyzed puppies. Pregnancy is an important physiological condition for the transmission of infectious agents to puppies and transplacental transmission may be epidemiologically relevant in the spread of these opportunistic agents.

Molecular detection of tick-borne protozoan parasites in a population of domestic cats in midwestern Brazil.

Some tick-borne pathogens that infect domestic cats have been considered emergent in veterinary medicine. Occurrences of Hepatozoon spp., Babesia spp. and Cytauxzoon spp. have been described in several regions of Brazil. This paper offers a comprehensive analysis of the 18S rRNA gene of a Hepatozoon sp. strain detected in domestic cats in the metropolitan area of Cuiabá, in Midwestern Brazil. Based on a molecular analysis, we detected the presence of Hepatozoon species circulating among cats in this region. The aforementioned strain is closely related to other isolates of H. felis detected in wild felids. Moreover, a phylogenetic analysis indicates that this genotype is grouped into a clade of 18S rRNA sequences previously described for the genus Hepatozoon in wild felids around the world. Hepatozoon felis strains detected in cats from Spain and Israel showed, respectively, 98% and 97% identity to our sequence and are clustered on a separate branch of the phylogenetic tree. This finding suggests a high diversity of Hepatozoon genotypes occurring in cats in Europe and South America. None of the analyzed cats were positive for Babesia spp. or Cytauxzoon spp. by PCR analysis.

Serological and molecular investigation of viral agents in freeliving jaguars of the Pantanal wetlands, state of Mato Grosso, Brazil / Investigação sorológica e molecular de agentes virais em onças-pintadas de vida livre no Pantanal de Mato Grosso, Brasil

This study investigates the exposure of free-living jaguars from two federal protected areas in the Pantanal of Mato Grosso, Brazil, to a variety viral agents. These viral agents, particularly causing zoonotic diseases, were analyzed using serological and molecular methods. None of the jaguars was positive by RT-PCR for the molecular detection of avian influenza and West Nile Fever (WNF). Only one animal was serologically positive for Eastern Equine Encephalitis (EEE) by virus neutralization test in VERO cell cultures, representing the first reported case of jaguar exposure to EEE virus. However, all the animals were negative for Western Equine Encephalitis (WEE) virus and Venezuelan Equine Encephalitis (VEE) virus. Eleven jaguars were tested by two tests for the detection of antibodies against rabies virus (Simplified Fluorescent Inhibition Microtest ­ SFIMT and Rapid Fluorescent Focus Inhibition Test ­ RFFIT), resulting in five positive animals, two animals in each test and one in both serological tests. Furthermore, three out of 14 samples subjected to the neutralization test were positive for antibodies against canine distemper virus (CDV), and 15 out of 17 samples subjected to the hemagglutination-inhibition test (HI) were positive for antibodies against canine parvovirus (CPV). In view of the findings of this study, it is unlikely that the viruses examined here represent a threat to the jaguar populations in this region.(AU)

Este estudo investigou a exposição de onças-pintadas de vida livre a agentes virais selecionados em duas unidades de conservação federais no Pantanal de Mato Grosso, Brasil. Para a análise desses agentes virais, a maioria de caráter zoonótico, foram utilizados métodos sorológicos e moleculares. Nenhuma das onze onças-pintadas examinadas foi positiva na técnica de real-time RT-PCR para a detecção molecular dos agentes da Influenza aviária e Febre do Nilo Ocidental (WNF). Somente um animal foi positivo sorologicamente para a o vírus da Encefalite Equina do Leste (EEE) pela Microtécnica de vírus neutralização em culturas de células VERO, sendo este o primeiro relato da exposição de onças-pintadas. Todos os animais examinados s foram negativos para o vírus da Encefalite Equina do Oeste (WEE) e Venezuelana (VEE). Amostras de soro colhidas de 11 onças-pintadas foram submetidas a adois testes distintos para a detecção de anticorpos contra o vírus da raiva (Teste Rápido de Inibição de Foco de Fluorescência ­ RFFIT e Microteste Simplificado de Inibição da Fluorescência - SFIMT), resultando em cinco animais positivos, dos quase dois positivos para cada teste e um positivo quando submetido aos dois testes sorológicos. Além disso, três das 14 amostras submetidas a técnica de soroneutralização foram positivas para a pesquisa de anticorpos contra o vírus da cinomose (CDV) e 15 amostras positivas das 17 analisadas para a pesquisa de anticorpos contra o parvovírus canino (CPV) foram identificadas pela técnica de Inibição da Hemaglutinação (HI). De acordo com os resultados deste estudo, é pouco provável que os agentes virais aqui analisados representem ameaça à população de onçaspintadas nesta região.(AU)