Avaliação da reação em cadeia pela polimerase para diagnóstico da leishmaniose visceral em sangue de cães

Canine and human visceral leishmaniasis is endemic in several States of Brazil, and it is associated with infected dogs and the presence of the vector. Aiming at using polymerase chain reaction as a diagnostic tool in dogs, we amplified a 120bp fragment from kDNA of Leishmania spp. by PCR in blood samples. The lower detection limit observed was 0.1 parasites per 500 mu L of blood, which is a highly satisfactory result. on the other hand, PCR evaluation in 166 blood samples of dogs from Poxoreo, MS, Brazil, resulted in 55% sensitivity and 66.3% specificity, considering indirect imunnofluorescent test as gold standard.

Phlebotomine (Diptera: Psychodidae) survey and some considerations about epidemiological aspects of canine leishmaniasis in Rio Claro city, São Paulo, Brazil.

Between 2000 and 2004, 170 sand flies were captured in urban or peri-urban areas of the Rio Claro city, SP, Brazil. Nyssomyia neivai, the main LTA vector species in São Paulo territory, corresponded to 124 specimens (72,94%) of the total number of captured insects. Incidence of American Cutaneous (LTA) and Visceral leishmaniasis (LVA) is increasing in human and dogs in São Paulo State. Rio Claro city is located in the center-east region of the State. During 2001 and 2003, two cases of alochtonus canine LVA and an autochthonous case of canine LTA were locally diagnosed. Results indicate the risk of LVA establishment in this region. Possible role of alternative vectors in leishmaniasis transmission is discussed.

Renal involvement in canine leishmaniasis: a morphological and immunohistochemical study

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Aplicação da técnica de imunofluorescência direta para o diagnóstico da leishmaniose visceral canina em aspirado de linfonodo

Recentemente, foco de leishmaniose visceral canina (CVL) foi descrito na região noroeste do Estado de São Paulo - Brasil. O Hospital Veterinário - UNESP - Araçatuba, no ano de 2.000, desenvolveu 60 testes citopatológicos de casos suspeitos de leishmaniose usando aspirado por agulha fina (FNA). Os esfregaços de linfonodo foram corados pelo método de Romanowsky (Diff-Quik®) e observados em microscopia de luz. Os casos positivos mostraram formas amastigotas típicas de Leishmania livres ou em vacúolos de macrófagos. Sinais citopatológicos de reatividade do sistema linfo-histiocitário com ausência de parasitos foram também observados. Com o objetivo de implementar o diagnóstico da CVL, detectando parasitos e material antigênico nos esfregaços, aplicou-se a reação de imunofluorescência direta (IFD) usando anticorpo policlonal anti-Leishmania produzido em camundongo. Comparamos o método de IFD com a pesquisa direta do parasito em esfregaços corados pelo método de Romanowsky. Dos 60 cães com sinais clínicos da doença, o exame direto foi positivo em 50% (n=30), duvidoso em 36,7% (n=22) e negativo com reatividade do linfonodo em 13,3% (n=8). Quando os linfonodos foram submetidos a reação de IFD observamos reação positiva em 93,3% (n=56) e reação negativa em 6,7% (n=4). Nossos resultados mostraram que a reação de IFD apresentou alta sensibilidade quando comparada a pesquisa direta do parasito pela coloração de Romanowsky. A reação de IFD pode ser um método útil para confirmar os casos duvidosos da doença, onde as formas amastigotas não são identificadas com facilidade.

Reposição de cães em área endêmica para leishmaniose visceral

Esse trabalho objetivou estimar a reposição de cães em área endêmica para leishmaniose visceral, onde a eutanásia de animais soropositivos é indicada como medida de controle, e avaliar os motivos que levaram a aquisição ou não de novos animais. Houve a reposição em 44,5% dos casos, principalmente devido à necessidade de companhia ou guarda. O principal motivo para a não-reposição foi o temor da leishmaniose visceral.

Canine cerebral leishmaniasis: Potential role of matrix metalloproteinase-2 in the development of neurological disease

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ocorrência de leishmaniose em gatos de área endêmica para leishmaniose visceral

Despite the description of several cases of feline leishmaniasis around the world, little information is available about the importance of the cat as a reservoir of the disease. The aim of the present study was to determine the occurrence of leishmaniasis in cats from an endemic area for visceral leishmaniasis in Brazil. Two hundred cats were included in this study. Infection was evaluated through the presence of amastigotes in stained smears from fine-needle aspirates of lymph nodes, bone marrow, spleen and liver, and by antibody reactivity against Leishmania chagasi using indirect ELISA. Our results showed a prevalence of infection in 14.5% (31/200) of the feline population studied, with 4% (8/200) of positivity by parasitological diagnosis and 11.5% (23/200) by serology.

Estudo da imunopatogenia das lesões no sistema nervoso central de cães naturalmente acometidos por leishmaniose visceral

Visceral leishmaniasis in dogs is described as a chronic disease whose main symptoms are progressive weigth loss, cachexy and dermatologic lesions. Recently, the disease has been associated to neurologic disorders. A total of 40 dogs with visceral leishmaniasis were divided into two groups. The first composed of dogs without neurological signs (n=30) and the second by dogs with neurological disorders (n=10). Brain samples were collected, stored in 10% buffered formalin and subjected to immunohistochemical examination for amastigotes forms of Leishmania (Leishmania) infantum chagasi, CD3+, CD4+ and CD8+ T lymphocytes and macrophages. Imunnohistochemistry evaluation revealed no amastigote forms of the parasite. CD3+ T lymphocytes were present in 24/30 (80%) dogs without neurological signs and in all dogs from the second group (p=0.0011). CD4+ and CD8+ were rarely observed, with CD4+ immunostaining in 10/40 (25%) dogs, from which half of them had neurological disease (p=0.0090). The presence of CD8+ was detected only in 4/10 (40%) dogs from neurological group (p=0.0021). Macrophages were detected in 38/40 (95%) dogs, without significant differences between groups (p=0.7664).

Neutrophil dysfunction varies with the stage of canine visceral leishmaniosis

Canine visceral leishmaniosis (CVL) causes a dependent-stage alteration in neutrophil oxidative metabolism. When production of reactive oxygen species (ROS) exceeds the antioxidant capacity of neutrophils, apoptosis is triggered, impairing the viability and function of these cells, which can predispose dogs to infection. However, the uremic condition observed in late-stage CVL can also alter the viability and function of human neutrophils. To more clearly understand this relationship, the apoptosis rate and oxidative metabolism of neutrophils from control dogs (n= 20) were compared to dogs in moderate (n= 15) and very severe (n= 15) stage CVL, classified according to LeishVet Consensus. To assess neutrophil oxidative metabolism, superoxide production was measured using the nitroblue tetrazolium reduction test (NBT) in isolated neutrophils. The apoptosis rate of neutrophils was estimated using the morphological method. Moderate-stage dogs presented increased superoxide production, while dogs with very severe stage CVL presented decreased superoxide production and an increase neutrophil apoptosis rate. Leishmaniosis causes differential neutrophil dysfunction according to disease stage. In moderate stage CVL, increased superoxide production is observed with no change in neutrophil viability. However, in very severe stage CVL, decreased superoxide production and increased apoptosis occur associated with uremia. © 2013 Elsevier B.V.

Morcegos como hospedeiros de Leishmania spp. em área endêmica para leishmaniose visceral

Pós-graduação em Ciência Animal - FMVA

Células CD4+FOXP3+ produzem lL-10 no baço de cães com leishmaniose visceral

Pós-graduação em Ciência Animal - FMVA

Avaliação histopatológica e imuno-histoquímica do miocárdio de cães com leishmaniose visceral: Fernando Azadinho Rosa. -

Pós-graduação em Ciência Animal - FMVA

Hipertensão arterial sistêmica e sua correlação com as lesões renais de cães naturalmente acometidos por leishmaniose visceral

Pós-graduação em Ciência Animal - FMVA

Correlação entre a presença de antígenos de Leishmania sp, linfócitos T CD3+ e B CD79+ e alterações histopatológicas no tecido renal de cães com leishmaniose visceral

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Expressão da glicoproteína-p e da mrp1 em tecidos de cães portadores de leishmaniose visceral

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)