220 resultados para CRYPTOSPORIDIUM-PARVUM


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

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

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

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A criptosporidiose, é a enfermidade de veiculação hídrica, possui como agravante a dificuldade de prevenção da contaminação ambiental e ausência de medidas terapêuticas eficazes. Com acentuada importância na bovinocultura, ocasiona inflamação e atrofia das vilosidades intestinais resultando em perda da superfície de absorção. Este estudo teve como objetivo realizar a caracterização molecular da infecção por Cryptosporidium spp. em bezerros do Município de Formiga, Minas Gerais. Um total de 300 amostras de fezes de bezerros holandeses, Nelore e sem raça definida saudáveis foram avaliadas pela técnica de coloração contraste negativo de verde malaquita e por meio da reação de Nested-PCR para amplificação de fragmentos de DNA da subunidade 18S do gene do RNA ribossômico. Ocorrência de 5,33% (16/300) pelo verde malaquita e 4,66% (14/300), pela PCR foi observada, sendo que nenhuma correlação foi verificada entre a positividade e as variáveis estudadas. Por meio da caracterização molecular foram identificadas as espécies Cryptosporidium andersoni e Cryptosporidium ryanae. Como conclusão, observou-se baixa ocorrência da infecção e eliminação de oocistos por Cryptosporidium spp, ausência de sinais clínicos nos animais, houve forte concordância entre os resultados obtidos por meio das duas técnicas utilizadas e pela caracterização molecular (Nested-PCR) foram diagnosticadas as espécies C. andersoni e C. ryanae, presentes em faixas etárias não relatadas na literatura. Estas duas espécies de Cryptosporidium supracitadas são descritas pela primeira vez, parasitando bovinos no estado de Minas Gerais.

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Pós-graduação em Medicina Veterinária - FCAV

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Parasitos do gênero Cryptosporidium pertencem ao filo Apicomplexa, com localização intracelular e extracitoplasmática obrigatória e se desenvolvem principalmente na superfície das células epiteliais de hospedeiros vertebrados. O cão, possível fonte de infecção humana, elimina oocistos fecais deste protozoário com grande potencial zoonótico no ambiente. O presente estudo teve como objetivo caracterizar molecularmente Cryptosporidium spp. obtidos de amostras fecais de filhotes caninos (naturalmente infectados). Um total de 200 cães foram examinados, sendo 100 machos e 100 fêmeas, 111 de padrão racial determinado e 89 sem raça definida (SRD). Destes, 81 animais, 43, 48 e 28 tinham até dois, de dois a três; de três a seis e de seis a doze meses, respectivamente. Conforme sua origem, os animais eram provenientes dos Municípios de Araçatuba e Votuporanga, SP, sendo que 126 eram de domicílios; 11 mantidos em Centros de Zoonoses; 50 de Pet Shops; 12 de um criatório e uma (0,5%) era errante e havia sido adotada. A ocorrência de Cryptosporidium spp. foi de 1% (2/200). Ambas eram fêmeas, SRD, com idade entre 60 e 90 dias. A de origem residencial apresentava fezes pastosas com coloração castanho claro e a outra, resgatada do CCZ, material fecal escurecido de consistência liquefeita. O sequenciamento dos fragmentos amplificados confirmou a presença de Cryptosporidium canis. A partir dos resultados obtidos neste trabalho, é possível concluir que 2% dos caninos analisados eram hospedeiros de C. canis

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

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Treatment of mice with the immunomodulating agent, Corynebacterium parvum (C. parvum), was shown to result in a severe and long-lasting depression of splenic natural killer (NK) cell-mediated cytotoxicity 5-21 days post-inoculation. Because NK cells have been implicated in immunosurveillance against malignancy (due to their spontaneous occurrence and rapid reactivity to a variety of histological types of tumors), as well as in resistance to established tumors, this decreased activity was of particular concern, since this effect is contrary to that which would be considered therapeutically desirable in cancer treatment (i.e. a potentiation of antitumor effector functions, including NK cell activity, would be expected to lead to a more effective destruction of malignant cells). Therefore, an analysis of the mechanism of this decline of splenic NK cell activity in C.parvum treated mice was undertaken.^ From in vitro co-culturing experiments, it was found that low NK-responsive C. parvum splenocytes were capable of reducing the normally high-reactivity of cells from untreated syngeneic mice to YAC-1 lymphoma, suggesting the presence of NK-directed suppressor cells in C. parvum treated animals. This was further supported by the demonstration of normal levels of cytotoxicity in C. parvum splenocyte preparations following Ficoll-Hypaque separation, which coincided with removal of the NK-suppressive capabilities of these cells. The T cell nature of these regulatory cells was indicated by (1) the failure of C. parvum to cause a reduction of NK cell activity, or the generation of NK-directed suppressor cells in T cell-deficient athymic mice, (2) the removal of C. parvum-induced suppression by T cell-depleting fractionation procedures or treatments, and (3) demonstration of suppression of NK cell activity by T cell-enriched C. parvum splenocytes. These studies suggest, therefore, that the eventual reduction of suppression by T cell elimination and/or inhibition, may result in a promotion of the antitumor effectiveness of C. parvum due to the contribution of "freed" NK effector cell activity.^ However, the temporary suppression of NK cell activity induced by C. parvum (reactivity of treated mice returns to normal levels within 28 days after C. parvum injection), may in fact be favorable in some situations, e.g. in bone marrow transplantation cases, since NK cells have been suggested to play a role also in the process of bone marrow graft rejection.^ Therefore, the discriminate use of agents such as C. parvum may allow for the controlled regulation of NK cell activity suggested to be necessary for the optimalization of therapeutic regimens. ^

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SCOPUS: le.j

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Diarrheal illness is responsible for over a quarter of all deaths in children under 5 years of age in sub-Saharan Africa and South Asia. Recent findings have identified the parasite Cryptosporidium as a contributor to enteric disease. We examined 9,348 cases and 13,128 controls from the Global Enteric Multicenter Study to assess whether Cryptosporidium interacted with co-occurring pathogens based on adjusted odds of moderate-to-severe diarrhea (MSD). Cryptosporidium was found to interact negatively with Shigella spp., with multiplicative interaction score of 0.16 (95% CI: 0.07 to 0.37, p-value=0.000), and an additive interaction score of -9.81 (95% CI: -13.61 to -6.01, p-value=0.000). Cryptosporidium also interacted negatively with Aeromonas spp., Adenovirus, Norovirus, and Astrovirus with marginal significance. Odds of MSD for Cryptosporidium co-infection with Shigella spp., Aeromonas spp., Adenovirus, Norovirus, or Astrovirus are lower than odds of MSD with either organism alone. This may reduce the efficacy of intervention strategies targeted at Cryptosporidium.

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High quality, pure DNA is required for ensuring reliable and reproducible results in molecular diagnosis applications. A number of in-house and commercial methods are available for the extraction and purification of genomic DNA from faecal material, each one offering a specific combination of performance, cost-effectiveness, and easiness of use that should be conveniently evaluated in function of the pathogen of interest. In this comparative study the marketed kits QIAamp DNA stool mini (Qiagen), SpeedTools DNA extraction (Biotools), DNAExtract-VK (Vacunek), PowerFecal DNA isolation (MoBio), and Wizard magnetic DNA purification system (Promega Corporation) were assessed for their efficacy in obtaining DNA of the most relevant enteric protozoan parasites associated to gastrointestinal disease globally. A panel of 113 stool specimens of clinically confirmed patients with cryptosporidiosis (n = 29), giardiasis (n = 47) and amoebiasis by Entamoeba histolytica (n = 3) or E. dispar (n = 10) and apparently healthy subjects (n = 24) were used for this purpose. Stool samples were aliquoted in five sub-samples and individually processed by each extraction method evaluated. Purified DNA samples were subsequently tested in PCR-based assays routinely used in our laboratory. The five compared methods yielded amplifiable amounts of DNA of the pathogens tested, although performance differences were observed among them depending on the parasite and the infection burden. Methods combining chemical, enzymatic and/or mechanical lysis procedures at temperatures of at least 56 °C were proven more efficient for the release of DNA from Cryptosporidium oocysts.