Serum proteins and electrophoretic profile in horses undergoing crotalid venom hyperimmunization.

Indisputably, the use of antivenoms for the treatment of snakebite envenoming is beneficial for the victims. However, there are few studies addressing the effect of long-term hyperimmunization in inoculated horses. It is known that the injection of snake venoms and adjuvants leads to local and systemic reactions in horses, but little is known about the response of inflammatory proteins. The aim of this study was to evaluate serum proteins and the electrophoretic profile of horses undergoing crotalid venom hyperimmunization. Twenty horses were divided into two groups: an inoculated group, comprising ten horses that were already being used for production of a Crotalus sp. antivenom, and a control group, comprising ten animals that had never been used for hyperimmunization. All animals were clinically healthy and without laboratory abnormalities. Total protein and albumin concentrations were measured in serum. Serum globulins were obtained by calculation. Plasma fibrinogen estimates were determined by the heat precipitation method. Serum proteinograms were obtained using agarose gel electrophoresis. The results revealed a significant increase in the concentrations of total serum proteins, globulins, and ß-globulins in the inoculated group, exceeding the reference values. There were slight increases in the α-1- and α-2-globulin subfractions in serum-producing horses, with no statistical significance. We also observed that horses used to produce hyperimmune plasma developed hypoalbuminemia, although the decrease in albumin production was not statistically significant. Our findings suggest that the continuous use of horses to produce crotalid antivenom may lead to a chronic inflammatory stimulus, with changes in plasma levels of inflammatory proteins.

Molecular detection of anaplasmataceae agents on dogs from the departament of Piura, Peru

This study aimed to study the prevalence of Anaplasmataceae organisms through the nested-PCR and phylogenetic analysis on domestic dogs in the Department of Piura, Peru. Two hundred and twelve canine blood samples were randomly collected on dogs from the central urban areas at the Piura Department in Peru. The extracted DNAs were tested, by nested-PCR based on 16SrRNA gene, to identify agents from Anaplasmataceae family. These results show that there was a prevalence of 18.5% (40/216) of positive dogs, 13.8% (30/216) for Ehrlichia canis, 7.4% (16/216) for Anaplasma platys and 0.1% (2/216) for Ehrlichia sp. confirmed by sequencing analysis. Co-positivity among Anaplasmataceae family species was present in 25% (10/40) of positive samples. There was a significant association among Anaplasmataceae family infection in dogs and the following variables: sex (p=0.034), presence of ticks (p=0.0001), and socio-economic status (p=0.001). There was no statistical association on the variables "living with other animals" and "age group" (p=0.1074). The partial sequences on the portion of the 16S rRNA gene, from positive samples for agents of Anaplasmataceae family demonstrated an identity of 97-100% with the isolated E. canis and A. platys obtained from the GenBank. This is the first study on infection by agents of Anaplasmataceae family in dogs in the Department of Piura, through molecular analysis

Rickettsial pathogens circulating in urban districts of Rio de Janeiro, without report of human Brazilian Spotted Fever.

Spotted fever group rickettsioses are emerging diseases. In some of these diseases, domestic dogs act as sentinels. Canine serological studies have demonstrated that rickettsial dispersion is concentrated in rural areas, seroprevalence being higher where human rickettsioses are endemic. In Rio de Janeiro, the Atlantic forest vegetation has been devastated by urbanization. In this context, we aimed to detect Rickettsia spp. in urban areas of the West Zone of Rio de Janeiro. Sera from 130 dogs were tested by Indirect Immunofluorescence Assay, and ticks collected from these dogs were tested by polymerase chain reaction. We found the rate of serological reactions against R. rickettsii and R. parkeri in our study area to exceed those of rural and non-endemic areas, highlighting the importance of dogs as urban sentinels. The possibility of contact with opossums and capybaras increased the chances of exposure to Rickettsia spp., reinforcing the hypothetical link between the landscape and the rickettsial wild cycle. Rhipicephalus sanguineus sensu lato was the tick most frequently observed. PCR-positive samples showed similarity with R. rickettsii and R. felis, an emerging pathogen rarely reported from ticks. We observed that rickettsiae circulate in urban places and ticks from indoor environments, which may be involved in bacterial epidemiology.

Spotted fever group rickettsial infection in dogs and their ticks from domestic-wildlife interface areas in southeastern Brazil.

Rickettsia rickettsii is the causative agent of Brazilian spotted fever (BSF), for which humans and dogs are both susceptible. Dogs are sentinels in serological surveys, however, canine disease is rarely reported. Therefore, we aimed to evaluate natural infection by spotted fever group (SFG) Rickettsia spp. in dogs and ticks collected from domiciles close to forest fragments, featuring domestic-wildlife interface areas. Samples from 115 dogs and 135 ixodids were assessed by polymerase chain reactions (PCR) targeting the gltA gene for Rickettsia spp. and the ompA gene for the SFG rickettsial species. One dog (0.87%; 1/115) was positive for R. rickettsii. This dog presented nonspecific laboratory and clinical abnormalities (thrombocytopenia, hyperproteinemia, lymph node enlargement, emaciation, anorexia, and lethargy). Rickettsia parkeri was identified in 2.96% (4/135) of the ticks (Amblyomma sculptum, A. aureolatum, and Rhipicephalus sanguineus). This study confirmed the presence of SFG bacteria in non-endemic and preserved locations, where domestic and wild populations interact. We reinforce the fact that the dog is susceptible to natural R. rickettsii infection. Although this is a rare finding, preventive measures should be taken against BSF in the studied areas. Finally, R. parkeri infection is possibly being demonstrated in A. sculptum for the first time.

Spotted fever group rickettsial infection in dogs and their ticks from domestic-wildlife interface areas in southeastern Brazil / Infecção por rickettsias do Grupo da Febre Maculosa em cães e carrapatos de áreas de interface doméstico-silvestre no sudeste Brasileiro

Abstract Rickettsia rickettsii is the causative agent of Brazilian spotted fever (BSF), for which humans and dogs are both susceptible. Dogs are sentinels in serological surveys, however, canine disease is rarely reported. Therefore, we aimed to evaluate natural infection by spotted fever group (SFG) Rickettsia spp. in dogs and ticks collected from domiciles close to forest fragments, featuring domestic-wildlife interface areas. Samples from 115 dogs and 135 ixodids were assessed by polymerase chain reactions (PCR) targeting the gltA gene for Rickettsia spp. and the ompA gene for the SFG rickettsial species. One dog (0.87%; 1/115) was positive for R. rickettsii. This dog presented nonspecific laboratory and clinical abnormalities (thrombocytopenia, hyperproteinemia, lymph node enlargement, emaciation, anorexia, and lethargy). Rickettsia parkeri was identified in 2.96% (4/135) of the ticks (Amblyomma sculptum, A. aureolatum, and Rhipicephalus sanguineus). This study confirmed the presence of SFG bacteria in non-endemic and preserved locations, where domestic and wild populations interact. We reinforce the fact that the dog is susceptible to natural R. rickettsii infection. Although this is a rare finding, preventive measures should be taken against BSF in the studied areas. Finally, R. parkeri infection is possibly being demonstrated in A. sculptum for the first time.

Resumo Rickettsia rickettsii é o agente causador da Febre Maculosa Brasileira (FMB), doença na qual humanos e cães são susceptíveis. Os cães são sentinelas nos inquéritos sorológicos, contudo, a doença canina é raramente descrita. Assim sendo, objetivou-se avaliar a infecção natural por Rickettsia spp. do Grupo da Febre Maculosa (GFM) em cães e carrapatos obtidos de domicílios próximos a fragmentos de mata, caracterizando áreas de interface doméstico-silvestre. Amostras de 115 cães e 135 ixodídeos foram avaliadas pela reação em cadeia da polimerase (PCR) tendo como alvo o gene gltA de Rickettsia spp. e o gene ompA das espécies do GFM. Um cão (0,87%; 1/115) foi positivo para R. rickettsii. Este animal apresentou alterações clínicas e laboratoriais inespecíficas (trombocitopenia, hiperproteinemia, linfonodos edemaciados, emagrecimento, anorexia e letargia). Rickettsia parkeri foi identificada em 2,96% (4/135) dos carrapatos (Amblyomma sculptum, A. aureolatum e Rhipicephalus sanguineus). Este estudo confirmou a presença de bactérias do GFM em locais preservados e não endêmicos, onde populações domésticas e silvestres interagem. Reforçamos o fato do cão ser susceptível à infecção natural por R. rickettsii. Embora este seja um achado raro, medidas preventivas devem ser tomadas contra a FMB nas áreas estudadas. Em última análise, a infecção por R. parkeri possivelmente está sendo demonstrada pela primeira vez em A. sculptum.

Linfocitose extrema associada à leucemia linfoblástica aguda (LLA) de células T em um cão jovem: relato de caso / Extreme lymphocytosis associated with acute T-cell lymphoblastic leukemia(ALL) in a young dog: case report

A leucemia linfoblástica aguda (LLA) é uma enfermidade de origem linfóide e consiste na proliferação de células neoplásicas na medula óssea. O objetivo desse trabalho é relatar o caso de um cão macho, sem raça definida, de apenas um ano de idade, atendido no Hospital Universitário de Medicina Veterinária Prof. Firmino Mársico Filho (HUVET) da Universidade Federal Fluminense (UFF) com queixa principal de inapetência e diarreia há três dias e que foi diagnosticado com essa neoplasia por meio da sintomatologia clínica, resultados do hemograma e do mielograma. O paciente apresentava valores exacerbados de linfócitos (553.094 células/µL), além de anemia, trombocitopenia, hipoalbuminemia e elevação da atividade das enzimas fosfatase alcalina e ALT. Foram observadas manchas de Gümprecht, linfócitos atípicos apresentando anisocitose, anisocariose, intensa basofilia citoplasmática e monócitos ativados. O mielograma apresentou também um aumento de linfócitos e contagem de linfoblastos superior a 30% na medula, confirmando o diagnóstico de leucemia linfoblástica aguda. Ademais, posteriormente, foi realizado exame de Reação em Cadeia de Polimerase (PCR) para rearranjos de receptores de antígenos e foi detectado clonalidade para linfócitos T. O animal foi submetido à quimioterapia (protocolo com ciclofosfamida, vincristina e prednisona), mas não resistiu à gravidade do quadro, vindo a óbito após a primeira sessão, pouco tempo após o diagnóstico.

Acute lymphoblastic leukemia (LLA) is a disease with a lymphoid origin and consists of the proliferation of neoplastic cells in the bone marrow. The aim of this study was to report the case of only one year old mixed breed male dog, attended at the University Hospital of Veterinary Medicine Prof. Firmino Mársico Filho (HUVET) from Universidade Federal Fluminense (UFF), with major complaint of inappetence and diarrhea three days ago and which was diagnosed with this neoplasm through clinical symptoms, complete blood count and myelogram results. The patient had increased values of lymphocytes (553,094 cells/μL), in addition to anemia, thrombocytopenia, hypoalbuminemia and elevated alkaline phosphatase and ALT activities. Gümprecht shadows, atypical lymphocytes presenting anisocytosis, anisocariosis, and severe cytoplasmic basophilia and activated monocytes were observed. Myelogram also showed an increase in lymphocytes and a lymphoblastic count greater than 30% in the marrow, confirming the diagnosis of LLA. In addition, polymerase chain reaction (PCR) for antigen receptor rearrangements was performed and clonality for T lymphocytes was detected. The animal underwent chemotherapy (protocol with cyclophosphamide, vincristine and prednisone), but did not withstand the severity of the disease, coming to death after the first session, shortly after diagnosis.

Linfocitose extrema associada à leucemia linfoblástica aguda (LLA) de células T em um cão jovem: relato de caso / Extreme lymphocytosis associated with acute T-cell lymphoblastic leukemia (ALL) in a young dog: case report

A leucemia linfoblástica aguda (LLA) é uma enfermidade de origem linfóide e consiste na proliferação de células neoplásicas na medula óssea. O objetivo desse trabalho é relatar o caso de um cão macho, sem raça definida, de apenas um ano de idade, atendido no Hospital Universitário de Medicina Veterinária Prof. Firmino Mársico Filho (HUVET) da Universidade Federal Fluminense (UFF) com queixa principal de inapetência e diarreia há três dias e que foi diagnosticado com essa neoplasia por meio da sintomatologia clínica, resultados do hemograma e do mielograma. O paciente apresentava valores exacerbados de linfócitos (553.094 células/µL), além de anemia, trombocitopenia, hipoalbuminemia e elevação da atividade das enzimas fosfatase alcalina e ALT. Foram observadas manchas de Gümprecht, linfócitos atípicos apresentando anisocitose, anisocariose, intensa basofilia citoplasmática e monócitos ativados. O mielograma apresentou também um aumento de linfócitos e contagem de linfoblastos superior a 30% na medula, confirmando o diagnóstico de leucemia linfoblástica aguda. Ademais, posteriormente, foi realizado exame de Reação em Cadeia de Polimerase (PCR) para rearranjos de receptores de antígenos e foi detectado clonalidade para linfócitos T. O animal foi submetido à quimioterapia (protocolo com ciclofosfamida, vincristina e prednisona) mas não resistiu à gravidade do quadro, vindo a óbito após a primeira sessão, pouco tempo após o diagnóstico.

Acute lymphoblastic leukemia (LLA) is a disease with a lymphoid origin and consists of the proliferation of neoplastic cells in the bone marrow. The aim of this study was to report the case of only one year old mixed breed male dog, attended at the University Hospital of Veterinary Medicine Prof. Firmino Mársico Filho (HUVET) from Universidade Federal Fluminense (UFF), with major complaint of inappetence and diarrhea three days ago and which was diagnosed with this neoplasm through clinical symptoms, complete blood count and myelogram results. The patient had increased values of lymphocytes (553,094 cells/µL), in addition to anemia, thrombocytopenia, hypoalbuminemia and elevated alkaline phosphatase and ALT activities. Gümprecht shadows, atypical lymphocytes presenting anisocytosis, anisocariosis, and severe cytoplasmic basophilia and activated monocytes were observed. Myelogram also showed an increase in lymphocytes and a lymphoblastic count greater than 30% in the marrow, confirming the diagnosis of LLA. In addition, polymerase chain reaction (PCR) for antigen receptor rearrangements was performed and clonality for T lymphocytes was detected. The animal underwent chemotherapy (protocol with cyclophosphamide, vincristine and prednisone), but did not withstand the severity of the disease, coming to death after the first session, shorly after diagnosis.

Bartonella henselae and Bartonella clarridgeiae infection, hematological changes and associated factors in domestic cats and dogs from an Atlantic rain forest area, Brazil.

Cats are considered main reservoir of Bartonella henselae, which is transmitted to other cats especially through Ctenocephalides felis fleas, and to humans through scratching and biting. Serra da Tiririca State Park (PESET) is an Atlantic Forest area that shelters a wide variety of endemic fauna. Recently, the park has been suffering due to irregular housing construction and domestic animal population that interacts with humans and wildlife. Given that surveillance policies for animals are part of the global Strategic Framework for One Health, the aim of this study was to detect Bartonella spp. DNA in cats and dogs, evaluating laboratory changes and associated factors. Blood samples of 124 dogs and 89 cats were collected for hematology and serum chemistry analysis. DNA was extracted and tested by conventional polymerase chain reaction (PCR) targeting a fragment of the citrate synthase (gltA) gene of Bartonella spp. with specific primers. Positive samples were sequenced to identify species. Bartonella henselae and B. clarridgeiae were detected in 24.7% of cats, being, for our knowledge, the first report of B. clarridgeiae in cats from Rio de Janeiro, Brazil. None of the samples obtained from dogs tested positive in the PCR assays. No statistical significance was observed in physical and laboratory exams. We suggest that cats that inhabit PESET can be considered sources of Bartonella sp. for other cats and humans. We highlight that infected cats did not present clinical or laboratory alterations. We alert for the need of care measures, avoiding scratch and bite, particularly in immunocompromised people.

Circulation of spotted fever group rickettsiae among dogs seropositive for Leishmania spp. in an urban area of Brazil.

INTRODUCTION: Dogs play an epidemiological role in several vector-borne diseases that affect human and animal health worldwide. We aimed to identify rickettsial circulation among dogs with canine visceral leishmaniasis (CVL) from a region endemic for both diseases. METHODS: CVL-seropositive dogs were screened for spotted fever group rickettsiae using an indirect immunofluorescence assay. RESULTS: Among the CVL-positive dogs, anti-Rickettsia rickettsii antibodies were identified in one asymptomatic and one oligosymptomatic dog. CONCLUSIONS: This study shows low circulation of antibodies to R. rickettsii in CVL-seropositive dogs. It is recommended that surveillance studies in dogs should continue in order to monitor this scenario.

Hematological evaluation of free-living golden-headed lion tamarins (Leontopithecus chrysomelas) from an Urban Atlantic Forest.

BACKGROUND: The Atlantic Forest where the animals were captured is surrounded by residences, so this close contact could favor the presence of disease-transmitting pathogens, putting the local population at risk. For these and other factors, it is important to perform laboratory tests enabling the performance of important diagnoses. METHODS: Blood samples of 268 golden-headed lion tamarins (Leontopithecus chrysomelas) inhabiting an Atlantic Forest area in Rio de Janeiro, Brazil, were processed for accessing age and sex influence in hematological parameters and to establish normative hematology values. RESULTS: Mean values of red blood cells, hematocrit, hemoglobin, and platelet count were significantly higher in adult males than in adult females. Adult animals had significantly higher mean neutrophil count, and young animals had higher averages than adults in lymphocyte values. Anisocytosis and platelet indices parameters were also provided for the first time. CONCLUSIONS: Averages presented can be used as hematological parameters for golden-headed lion tamarins.

Circulation of spotted fever group rickettsiae among dogs seropositive for Leishmania spp. in an urban area of Brazil

Abstract INTRODUCTION: Dogs play an epidemiological role in several vector-borne diseases that affect human and animal health worldwide. We aimed to identify rickettsial circulation among dogs with canine visceral leishmaniasis (CVL) from a region endemic for both diseases. METHODS: CVL-seropositive dogs were screened for spotted fever group rickettsiae using an indirect immunofluorescence assay. RESULTS: Among the CVL-positive dogs, anti-Rickettsia rickettsii antibodies were identified in one asymptomatic and one oligosymptomatic dog. CONCLUSIONS: This study shows low circulation of antibodies to R. rickettsii in CVL-seropositive dogs. It is recommended that surveillance studies in dogs should continue in order to monitor this scenario.

Assessment of the diagnostic performance of four methods for the detection of Giardia duodenalis in fecal samples from human, canine and feline carriers.

Enteric parasitic diseases including giardiasis are of public health concern. Different methods are available for the diagnosis of this parasitic infection in fecal samples such as the identification of protozoan cysts and trophozoites by light microscopy, detection of specific antigens by ELISA, and amplification of DNA fragments by PCR. The present study aimed at assessing the performance of four laboratory tests for the detection of Giardia duodenalis in fecal specimens from three different host species with a previous diagnosis of giardiasis; canine, feline and human patients provided new stool samples to be retested for Giardia before initiating treatment with antiprotozoal drugs. For this purpose, triplicate fecal specimens from 54 humans, 24 dogs and 18 cats living in the city of Niterói, RJ, southeast Brazil, were analysed by light microscopy, ELISA, immunochromatography, and nested PCR. The centrifugal-flotation method detected Giardia cysts in 89.6% (86/96) of the fecal samples. The protozoan parasite was detected via immunochromatography in 87.5% (84/96) of these samples. Giardia was detected by ELISA in 69.8% (67/96) of the stool specimens from carriers with a previous diagnosis of Giardia infection. Giardia was detected by PCR in only 39.6% (38/96) of the fecal specimens. Based on these findings, we suggest that, among the four assays that were used in this study, the zinc sulphate flotation technique (Faust et al., 1939) is the best diagnostic assay in terms of sensitivity and specificity to detect G. duodenalis on serially collected samples from dogs, cats and humans.

Novel record of a tick Amblyomma sculptum with detection of piroplasm Rangelia vitalii DNA.

Rangelia vitalii is the protozoan responsible for a febrile tick-borne disease known as "Nambiuvú" or "bleeding plague." The only competent vector of this protozoan recognized in Brazil is the tick Amblyomma aureolatum. The aim of this study was to identify R. vitalii using molecular biology in ticks parasitizing Cerdocyon thous from Tijuca and Serra dos Órgãos National Parks, which are areas of the Atlantic Rainforest biome in the municipality of Rio de Janeiro. After taxonomic identification, ticks were crushed and DNA was extracted and examined by polymerase chain reaction (PCR) to verify amplification of a fragment of the 18S rRNA gene of piroplasms. Samples identified as positive in the PCR analysis were subjected to sequencing, which revealed high identity with sequences of R. vitalii. This is the first report of parasitizing nymphs of A. sculptum infected by R. vitalii under natural conditions.

Behavior of red cell distribution width (RDW) in a population of anemic and non-anemic dogs from an ambulatory care unit

O objetivo deste estudo foi avaliar o comportamento do índice de anisocitose na interpretação do hemograma de cães anêmicos e não anêmicos. Atualmente, contadores hematológicos automatizados veterinários fornecem alguns índices que não são calculados quando o hemograma é confeccionado manualmente. A saber: RDW-CV (Amplitude de distribuição de hemácias como coeficiente de variação) e RDW-SD (Amplitude de distribuição de hemácias como desvio-padrão). Tais índices levaram a uma nova abordagem do paciente com anemia permitindo que a anisocitose fosse mensurada de uma forma quantitativa. Foram avaliados 454 hemogramas processados no período de setembro de 2009 a março de 2011. Foram calculados média e desvio padrão e uma análise de variância foi realizada. Os cães anêmicos (125/454) apresentaram altos valores de RDW enquanto que, em animais não anêmicos, esse índice permaneceu dentro dos valores de normalidade previamente estabelecidos. O índice também estava alto em animais com anisocitose observada à microscopia. Houve uma diferença significante entre os índices dos animais que apresentaram Volume Globular Médio (VGM) elevado e normal. Concluiu-se que os índices RDW-CV e RDW-SD foram sensíveis a pequenas variações no tamanho das hemácias e na heterogeneidade, sendo mais precisos que a observação microscópica para se detectar anisocitose e mais sensível que VGM para variação eritrocitária.

The aim of this study was to evaluate the behavior of the red cell distribution width (RDW) in the interpretation of the Complete Blood Count (CBC) from anemic and non-anemic dogs. Currently, automated veterinary blood cell analyzers provide hematologic indices not calculated in manually performed CBC, such as Red Blood Cell Distribution Width measured by Variation Coefficients (RDW-CV) and Red Blood Cell Distribution Width measured by Standard Deviation (RDW-SD), which has led to new of September 2009 to March 2011. Mean and standard deviation were calculated and variance analysis was performed. Anemic dogs (125/454) presented higher values for RDW indices. In non-anemic animals, these indices were in accordance with the normal values previously established. Both indices were higher in dogs with microscopic observation of anisocytosis. There was a significant difference among índices of the animals which showed normal and increased Mean Corpuscular Volume (MCV). We concluded that RDW-CV and RDW-SD indices were sensitive to small variations on erythrocyte size and heterogeneity, being more accurate than microscopic observations to detect anisocytosis and more sensitive than MCV to erythrocyte variation.

Brazilian Spotted Fever with an Approach in Veterinary Medicine and One Health Perspective.

There is increasing interaction between man and pathogens transmitted by arthropods, especially by ticks. It is on this background that a holistic approach stands out, for the sake of Public Health. Brazilian Spotted Fever is an endemic disease at the country's southeast, with Amblyomma sculptum as its major contributor, followed by A. aureolatum and potentially Rhipicephalus sanguineus. Dogs have been considered sentinels, and in some areas the disease in dogs can precede human disease. Considering the importance of this disease for human health, the serological evidence in dogs, and the transmission of ticks between dogs and their owners, this review aimed to elucidate the importance of the epidemiological investigation, the diagnosis in dogs, and the role of veterinarians in Public Health to control vector-borne zoonotic diseases. We encourage veterinarians to include this rickettsial infection in the diagnosis of febrile diseases of common occurrence in dogs.