Made available in DSpace on 2014-07-29T15:29:13Z (GMT). No. of bitstreams: 1 Dissertacao Solimar A de Oliveira.pdf: 1870870 bytes, checksum: 5c7849377c8610cb5cbc8b8d4905a909 (MD5) Previous issue date: 2012-02-10 / Introduction: The chronic renal insufficiency is in between the transition epidemiologist illness, being able to be affected by the enteric opportunistic parasites for representing a population of immunosuppressed. Catalogued as emergent agents of opportunistic character, protozoan disease is responsible for important morbi-mortality rates, but little recognized on the part of the professionals of health and for the shortage of specialized laboratories in its diagnostics. They are caused mainly by protozoan, as the Blastocystis hominis, Cryptosporidium sp, Isospora belli, Cyclospora cayetanensis, amongst others. Objectives: Mapping world-wide studies through a systematic revision of literature concerned to the detection of these protozoan in hemodialysis patients. And, besides, to identify enteric opportunistic agents in immunosuppressed children with chronic nephropathies who were submitted to hemodialysis and also children patients who don t have chronic nephropathies, in the Clinical Hospital /UFG. Methods: The theoretical part, represented by the systematic revision of literature, was elaborated from standardized forms on the selection of scientific articles available in the Virtual Library in Health. This work, concerning the experimental part, was built in the period of October of 2009 to May of 2011 with analysis of the epidemiologic profiles of the patients and laboratorial detection of opportunistic enteroparasitosis in 229 fecal samples of 26 hemodialysis children (test-group) and of 59 children who haven t chronic nephropathies (control group), from the Hospital of the Clinics of the Federal University of Goiás. For further detection of the oocysts of coccids (Cryptosporidium sp, Cyclospora cayetanensis and Isospora belli), microscopic examination was used direct (fresh).It was also used the methods of Hoffman, Pons and Janer, Ridley or of concentration in formalin 10% - Acetate of Ethyl, Coloration of Kinyoun (hot), and, Ziehl-Neelsen (modified). With regard to the diagnosis of Blastocystis hominis, it was used the microscopic examination direct (fresh) and the technique of Coloration of Nair - Blue of Methylene. Results: In the systematic revision of literature, nine articles had been selected, and from the interpretation on these studies, the presence of enteric opportunistic protozoan in fecal samples of hemodialysis patients was evidenced. In the experimental part, the detection of protozoan for patients, in the test group and in the control group was, respectively, of: Blastocystis hominis in 9 (34,6%) and 13 (22%); Giardia lamblia in 3 (11,5%) and 2 (3,4%); Endolimax nana in 9 (34,6%) and 9 (15,3%); Entamoeba coli in 3 (11,5%) and 2 (3,4%). And still it had been detected only in the test group: Cryptosporidium sp in 1 (3,8%) patient and Entamoeba histolytica/dispar in 3 (11,5%). Regarding the quantitative analysis of fecal samples, it was collected 115 samples of the group of hemodialysis children and 114 samples of the group of children who don t have chronic nephropathies. The results gotten in this comparison had designated the presence of oocysts of intestinal protozoan in the test group and in the control group. Respectively, we have: Blastocystis hominis in 24 (20,87%) and 16 (14,04%) samples; Giardia lamblia in 3 (2,61%) and 2 (1,75%) samples; Endolimax nana 15 (13,4%) and 9 (7,89%) samples. Besides, it had been detected only in the test group: Cryptosporidium sp in 1 (0,87%) sample and Entamoeba histolytica/dispar in 3 (2,61%). With regard to the diarrheal feces analysis, it was detected in test group 1 sample (0,87%), and in the control group, 8 (7.02%). Conclusion: These findings demonstrate that such agents are present in our environment, reinforcing isolated infections or associates, between them or ahead of other opportunistic enteric parasites, providing a risk for the population of hemodialysis patient. They still disclose the urgency of an implantation of specialized laboratories with specific detection techniques of these infectum-parasitic agents. / Introdução: A insuficiência renal crônica está entre as doenças de transição epidemiológica, podendo ser afetada pelas parasitoses entérico oportunistas por representar uma população de imunossuprimidos. Catalogadas como agentes emergentes de caráter oportunista, as protozooses são responsáveis por importantes índices de morbi-mortalidade, mas pouco reconhecidas por parte dos profissionais de saúde e pela escassez de laboratórios especializados em seus diagnósticos. São causadas, principalmente, por protozoários como o Blastocystis hominis, Cryptosporidium sp, Isospora belli, Cyclospora cayetanensis, dentre outros. Objetivos: Mapear estudos mundiais mediante revisão sistemática da literatura quanto à detecção destes protozoários em pacientes hemodialisados. Identificar agentes entéricos oportunistas em crianças imunossuprimidas com nefropatias crônicas e submetidas à hemodiálise e em crianças não portadoras de nefropatias crônicas, no Hospital das Clínicas/UFG. Métodos: A parte teórica, representada pela revisão sistemática da literatura, foi elaborada a partir de formulários padronizados para a seleção de artigos científicos existentes na Biblioteca Virtual em Saúde. Este trabalho, no que tange à parte experimental, foi realizado no período de outubro de 2009 a maio de 2011, com análise do perfil epidemiológico dos pacientes e detecção laboratorial de enteroparasitoses oportunistas em 229 amostras fecais de 26 crianças hemodialisadas (grupo teste) e de 59 crianças não portadoras de nefropatias crônicas (grupo controle), procedentes do Hospital das Clínicas da Universidade Federal de Goiás. Para a detecção dos oocistos de coccídeos (Cryptosporidium sp, Cyclospora cayetanensis e Isospora belli), foram utilizados exames microscópicos diretos a fresco, bem como os métodos de Hoffman, Pons e Janer, Ridley ou de concentração em formalina a 10% Acetato de Etila, Coloração de Kinyoun a quente e ainda, Ziehl-Neelsen modificado. Já para o diagnóstico de Blastocystis hominis, foram utilizados exames microscópicos diretos a fresco e a técnica de Coloração de Nair Azul de Metileno. Resultados: Na revisão sistemática da literatura foram selecionados nove artigos e a partir da interpretação desses estudos foi constatada a presença de protozoários entéricos oportunistas em amostras fecais de pacientes hemodialisados. Na parte experimental, a detecção de protozoários por pacientes, no grupo teste e no grupo controle, foi de: Blastocystis hominis em 9 (34,6%) e 13 (22%); Giardia lamblia em 3 (11,5%) e 2 (3,4%); Endolimax nana em 9 (34,6%) e 9 (15,3%); Entamoeba coli em 3 (11,5%) e 2 (3,4%), respectivamente. Ainda, foram detectados no grupo teste: Cryptosporidium sp em 1 (3,8%) paciente e Entamoeba histolytica/dispar em 3 (11,5%). Quanto à análise quantitativa de amostras fecais, foram coletadas 115 amostras fecais do grupo de crianças hemodialisadas e 114 amostras fecais do grupo de crianças não portadoras de nefropatias crônicas. Os resultados obtidos nessa comparação assinalaram a presença de cistos e oocistos de protozoários intestinais no grupo teste e no grupo controle. Nos referidos grupos teste e controle foram encontrados cistos de Blastocystis hominis em 24 (20,87%) e 16 (14,04%) amostras; Giardia lamblia em 3 (2,61%) e 2 (1,75%) amostras; Endolimax nana 15 (13,4%) e 9 (7,89%) amostras, respectivamente. Além disso, foram detectados no grupo-teste: Cryptosporidium sp em 1 (0,87%) amostra e Entamoeba histolytica/dispar em 3 (2,61%). Em relação à consistência das fezes, foi detectada fezes diarréicas em 1 (0,87%) amostra do grupo-teste e 8 (7,02%) do grupo controle. Conclusão: Estes achados demonstram que tais agentes estão presentes em nosso meio ambiente, potencializando infecções isoladas ou associadas, entre eles ou diante de outros parasitos entéricos oportunistas, proporcionando um risco para a população de hemodialisados. Revelam ainda, a premência de implantação de laboratórios especializados com técnicas específicas de detecção destes agentes infecto-parasitários.

Foodborne illnesses are a significant public health challenge in the United States, with an estimated 9.4 million illnesses annually attributed to the consumption of contaminated food, of which 59% are estimated to be caused by viruses, 39% by bacteria and 2% by parasites. Timely detection and identification of the pathogens causing foodborne outbreaks is vital for the implementation of outbreak control strategies, allowing public health officials to prevent additional illnesses and maintain confidence in the food supply. Public health laboratories employ a variety of traditional and molecular testing techniques to identify foodborne outbreak etiologic agents. One technology is the Luminex XMap® microsphere system, which is also marketed as the Bio-Plex™ 200. This platform has a multiplexing capability with the potential to simultaneously detect up to 100 targets in one reaction. The studies described here show that the combination of two Bio-Plex assays with real-time virus assays and one extraction method provides a flexible foodborne outbreak screening algorithm that potentially identifies an outbreak-associated pathogen on the first day of specimen submission and aids in focusing confirmatory laboratory testing. In these studies, two microsphere-based assays were designed for use on the Bio-Plex 200 system as screening assays for the detection of four enteric protozoa (Giardia intestinalis, Cyclospora cayetanensis, Cryptosporidium parvum, Entamoeba histolytica) and six virulence determinants of shiga toxin-producing Escherichia coli (STEC), enterotoxigenic Escherichia coli (ETEC), enteroinvasive Escherichia coli (EIEC) and Shigella spp. Precision and limits of detections were established for both assays. The sensitivity and specificity of the protozoan assay as compared to reference methods ranged from 81.25% to 100% for most targets, while sensitivity for the E. histolytica target was 42.86%. Sensitivity and specificity for the bacterial assay was 100% as compared to reference methods. However, cross-reactivity of the protozoan assay E. histolytica target with E. dispar and of the bacterial assay uidA target with enteropathogenic E. coli strains was noted. Additionally, real-time detection of norovirus and rotavirus nucleic acids extracted with the QIAamp DNA Stool Mini Kit was statistically comparable to detection when extracted with the Ambion® MagMAX™-96 Viral RNA Isolation Kit combined with the KingFisher® Magnetic Particle Processor.