Dissertations / Theses on the topic 'Intestinal and parasitic'

Orientador: Regina Maura Bueno Franco<br>Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia<br>Made available in DSpace on 2018-08-20T10:49:21Z (GMT). No. of bitstreams: 1 Ribeiro_DaniloCarlos_M.pdf: 1808064 bytes, checksum: 36f0510d37feb2ae734deffdba6cf10f (MD5) Previous issue date: 2011<br>Resumo: As parasitoses intestinais representam um grave problema de Saúde Pública em todo o mundo, com prevalências variáveis em cada região, sendo que fatores socioeconômicos e ambientais corroboram para manutenção ou aumento do número de casos. Os trabalhadores rurais canavieiros, devido as condições as quais são submetidos durante o trabalho sofrem exposições diárias aos agentes parasitários. Eles formam um grupo populacional cuja saúde é negligenciada devido a escassez de informações na literatura. Os objetivos deste trabalho foram: avaliar a prevalência das infecções parasitárias intestinais entre trabalhadores rurais de duas principais regiões canavieiras do Estado de São Paulo (Ribeirão Preto e Campinas) destino de milhares de migrantes oriundos de diferentes regiões do Brasil no período de safra da cana de açúcar; avaliar comparativamente os métodos de coloração de Ziehl Neelsen modificada e Auramina O, para detecção dos oocistos de coccidioses intestinais e, as técnicas de Hoffman, Pons e Janer (H.P.J.) e Coprokit ® para detecção de ovos de helmintos. Foram analisadas 3 amostras fecais de 617 indivíduos, as quais foram processadas pelos métodos de sedimentação espontânea, Faust et al., Rugai et al., Willis et al., Coprokit ®, e pelos métodos de coloração Auramina- O e Ziehl Neelsen modificada, mediante emprego de técnica de centrifugo - concentração a 500X g e, aplicação de um questionário semi- estruturado para avaliar condições sócioeconômicas e epidemiológicas. A prevalência geral de parasitos intestinais e comensais foi de 30,47 %, sendo que o protozoário mais encontrado foi Endolimax nana (12,16 %) e entre os helmintos detectou-se com maior frequência os ancilostomídeos (2,43 %). Dentre os exames positivos, foi verificada maior ocorrência de monoparasitismo (23,82 %) e de protozoários (26,58 %). Conclui-se que a baixa prevalência encontrada para parasitoses intestinais neste estudo se deve as melhorias no perfil socioeconômico e epidemiológico dos trabalhadores canavieiros e entre os exames positivos não houve diferença significativa na detecção de oocistos de Cryptosporidium spp. pelas técnicas de Ziehl Neelsen modificada e Auramina O e, a técnica de sedimentação espontânea se mostrou mais eficiente para detecção de ovos de helmintos em comparação ao método de Coprokit ®<br>Abstract: Intestinal parasitic diseases represent a serious public health problem worldwide, with prevalence rates varying in each region, and socioeconomic and environmental factors to corroborate maintaining or increasing the number of cases. Rural workers in sugar cane plantations, due to weather conditions and which are submitted during the rural work, and suffer daily exposures to parasitic agents. They form a population group whose health is neglected due to lack of information in the literature. Our objectives were: assess the prevalence of intestinal parasitic infections among rural workers in two major sugar cane growing regions of São Paulo (Ribeirão Preto and Campinas) fate of thousands of migrants from different regions of Brazil from sugar cane crop; benchmarking the methods of Ziehl Neelsen modified and Auramina O for detection of oocysts of intestinal coccidia and the techniques of Hoffman, Pons and Janer (H.P.J.) and Coprokit ® for detection of helminth eggs. Three fecal samples were analyzed from 617 individuals, which were processed by the methods of spontaneous sedimentation, Faust et al., Rugai et al., Willis et al. Coprokit ®, and the staining methods Auramina O and Ziehl Neelsen modified, by use of centrifugal technique - the 500Xg concentration and, application of a semi-structured survey to assess the socioeconomic and epidemiological conditions. The overall prevalence of parasitism was 30.47%, and the most frequent protozoan was Endolimax nana (12,16%) and among the helminthes were detected more frequently the hookworm (2,43%). Among the positive tests, there was verified a higher incidence of monoparasitism (23,82%) and protozoa (26,58%). It is concluded that the low prevalence for intestinal parasites in this study is due to improvements in socio-economic and epidemiological profile of the sugarcane workers and among the positive tests there was no significant difference in detection of oocysts Cryptosporidium spp. by modified Ziehl Neelsen technique and Auramina O, and the spontaneous sedimentation technique was more efficient for detection of helminth eggs in comparison to the method of Coprokit ®<br>Mestrado<br>Parasitologia<br>Mestre em Parasitologia

The parasitic nematodes Ancylostoma caninum and Oesophagostomum radiatum were collected and analyzed for intestinal inorganic granules. Three means of identification were utilized to determine the composition granules, including birefringence, x-ray diffraction and energy dispersive spectrometric (EDS) analysis. Initial x-ray diffraction results of the two worms showed a calcium sulfide presence within the worms. Closer examination of the granules within Ancylostoma caninum however, utilizing EDS analysis revealed their composition to be zinc sulfide. These results concur with those of Rogers (1940) and Clark (1956) who found zinc sulfide granules in several species of Strongylus. The ZnS granules seem to be a result of a detoxification function that binds excess zinc and sulfhydryl groups present from the ingestion and breakdown of dietary blood meals.

Made available in DSpace on 2014-10-09T12:49:59Z (GMT). No. of bitstreams: 0<br>Made available in DSpace on 2014-10-09T14:02:43Z (GMT). No. of bitstreams: 0<br>Dissertacao (Mestrado)<br>IPEN/D<br>Instituto de Pesquisas Energeticas e Nucleares - IPEN/CNEN-SP

Intestinal parasitic infections (IPI's) are considered to be a public health problem of global importance by the World Health Organization. The present epidemiologic survey study investigated the prevalence, risk factors, and consequences of pathogenic IPI's on the growth, nutrition and psychomotor development of 244 Ecuadorian children aged 0.2-14 years. The study was conducted in five rural hamlets located in a tropical rainforest area in northwest Ecuador. The study data were obtained by means of a structured questionnaire, a developmental screening examination, anthropometry, and lab analysis of blood and fecal samples. Data analysis was conducted using appropriate bivariate and multivariate statistical techniques. The study results revealed that 90% of the child subjects were infected with at least one pathogenic IPI species. Fifty-one percent were identified with helminthic infections, 37.6% with protozoal infections, and 21.4% were infected with both. The most common intestinal parasites detected were Ascaris lumbricoides (39.7%), Giardia intestinalis (25.2%), Trichuris trichiura (19.7%), Entamoeba histolytica/dispar (18.5%), Blastocystis hominis (13.3%), and Ancylostoma duodenale (1.7%). The prevalence of growth stunting (40%) and iron-deficiency anemia (26%) also was high. Children infected with Giardia exhibited a risk for stunted growth that was twice that of their non-infected counterparts (51.7% vs. 33.1%; OR=2.16, 95% C.I.= 1.13-4.15; p= 0.01). They also had significantly reduced mean blood hemoglobin levels compared to non-infected children 11.8 + 1.5.g/dL vs. 12.2 + 1.4g/dL; p= 0.023) but the proportion with iron-deficiency anemia was slightly but not significantly increased (29.4% vs. 24.3%). The characteristic most consistently associated with risk for pathogenic protozoal IPI's was a high density of domestic animals living in and around the home. Children who lived in such households had a risk for infection that was 2-5 times greater than others. This suggests that domestic animals were important reservoirs for IPI infection in the child group studied. Contrary to the a priori hypothesis, no gender, ethnic, nor age differences in infection risk were identified except for Trichuris infection, which was reduced in younger children contrary to expectations. Mass or targeted chemotherapy combined with health education and promotion are needed to reduce the cycle of infection and re-infection and the negative impact of these on child growth and iron status. Health education and promotion messages can be incorporated into other types of programs already in place in local schools and by the Ecuadorian Ministries of Public Health, Education, and Social Welfare and other agencies.<br>Master of Science

Dissertação de Mestrado Integrado em Medicina Veterinária<br>A presente dissertação é o resultado do estágio realizado na Faculdade de Ciências Agrárias e Veterinárias da Universidade Estadual Paulista “Júlio de Mesquita Filho”, Campus de Jaboticabal, São Paulo, Brasil. É composta por uma descrição resumida das actividades desenvolvidas durante o estágio, exposição breve da casuística acompanhada, seguida de uma revisão bibliográfica do tema proposto. Esta revisão incide sobre as funções do intestino na nutrição do animal e destaca o papel essencial da dieta na nutrição do intestino. Estuda-se a importância do intestino na imunidade, relacionando os mecanismos de resistência a parasitas intestinais (endo e extracelulares) em cães. No âmbito do tema escolhido, são referidos os efeitos específicos das deficiências de nutrientes a nível molecular ou de produção de citoquinas específicas. Há muitas pesquisas que demonstram que a má nutrição e a infecção ocorrem em conjunto. Não podem ser feitas generalizações sobre os efeitos de diversos nutrientes sobre os vários componentes da resposta imune, e a falta de compreensão da base de imunidade funcional contra nemátodes, torna difícil identificar as deficiências nutricionais que deveriam ser de maior preocupação. Neste estudo, o foco é centrado no intestino, que é o local da digestão e absorção de nutrientes e de permanência da maioria dos parasitas. Como complemento do tema, procede-se ao estudo dos aspectos nutricionais de sete casos clínicos acompanhados pelo autor com a respectiva discussão. Por fim, salientam-se as conclusões obtidas. Em Portugal, o autor realizou um inquérito a Médicos Veterinários sobre Nutrição Clínica, demonstrando-se que é uma área subvalorizada no nosso país. É abordada a importância de profissionais nesta área e de cursos de Nutrição Clínica para os veterinários. O tecido linfóide associado ao intestino é o maior componente do sistema imunitário do organismo. Há uma relação dinâmica entre nutrição, imunidade e doença e esta área interdisciplinar de investigação necessita de uma maior cooperação entre veterinários, parasitologistas, nutricionistas, imunologistas, biólogos moleculares e profissionais de saúde pública.<br>ABSTRACT - GUT NUTRITION, INTESTINAL IMMUNITY AND RESISTANCE TO INTESTINAL PARASITES OF DOGS - This thesis is the result of the training held at the Faculty of Agriculture and Veterinary Sciences, Universidade Estadual Paulista “Júlio de Mesquita Filho”, Jaboticabal Campus, São Paulo, Brazil. The description of the activities undertaken during the training, brief overview of the casuistic, followed by a literature review of the proposed theme are presented. This review focuses on the functions of the gut in animal nutrition and highlights the essential role of diet in the nutrition of the intestine. The importance of gut immunity and the relationship between mechanisms of resistance to intestinal parasites (endo and extracellular) in dogs are mentioned. As a complement of the subject, a study of nutritional aspects of seven clinical cases are referred and followed by discussion and conclusions. Considering the aim of the present work the specific effects of nutrient deficiencies at the molecular level or production of specific cytokines are highlighted. There are many studies showing that malnutrition and infection occur together. No generalizations can be made on the effects of various nutrients on the various components of the immune response. The knowledge of functional immunity basis against nematode is needed to clear identify nutritional deficiencies. In this study, the focus is centred in the intestine, an organ were absorption and digestion of nutrients as well as localization of a large number of parasites do occur. In Portugal, the author conducted a questionnaire to Veterinarians about Clinic Nutrition. The results allowed to conclude that this area is undervalued in our country and it must be taken into account the need of experts and training courses in clinical nutrition for veterinarians. The lymphoid tissue associated with the intestine is the major component of the body's immune system. There is a dynamic relationship between nutrition, immunity and disease, and this interdisciplinary research requires greater cooperation between veterinarians, parasitologists, nutritionists, immunologists, molecular biologists and public health professionals.

Orientadores: Manzélio Cavazzana Júnior, Regina Maura Bueno Franco<br>Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia<br>Made available in DSpace on 2018-08-19T19:07:23Z (GMT). No. of bitstreams: 1 Malta_RobertoCarlosGrassi_D.pdf: 3672169 bytes, checksum: 25ac1445db6b62e9892dee0a7eea948b (MD5) Previous issue date: 2011<br>Resumo: As infecções parasitárias são um dos principais problemas de saúde pública, apresentando-se de forma endêmica em diversas áreas do Brasil. Podem apresentar estreita relação com fatores sócio-demográficos e ambientais, tais como: precárias condições socioeconômicas, consumo de água contaminada, deficiente estado nutricional dos indivíduos e outros, sendo frequentemente a população infantil a mais atingida. Com o objetivo de investigar a prevalência de parasitas intestinais em populações indígenas e populações carentes e os fatores-chave envolvidos na epidemiologia de enteroparasitoses, foi realizado levantamento enteroparasitológico em moradores de 02 reservas indígenas Reserva Bororó/MS e Reserva Xingu/MT (tribos Kayabí e Juruna) -, e também em moradores de 02 cidades - Pontes e Lacerda/MT e Ibateguara/AL. A coleta de dados foi realizada de 2002 a 2009. Foram analisadas 2754 amostras de fezes pelos métodos de Faust, Hoffman, Kato-Katz, Rugai, Direto e Ziehl-Neelsen modificado. Foram obtidos dados pessoais e parâmetros socioeconômicos. Observou-se a presença de 73% de enteroparasitas na reserva indígena de Dourados, 62,77% na reserva indígena do Xingu, 52,61% no município de Pontes e Lacerda/MS e 67,42% no município de Ibateguara. As espécies de maior prevalência no sexo masculino foram Entamoeba coli (22,5%), Giardia duodenalis (11,6%), Entamoeba histolytica (13,9%) e Ascaris lumbricoides (13,6%). No sexo feminino foram Entamoeba coli (24,1%), Giardia duodenalis (8,8%), Entamoeba histolytica/díspar (13,8%) e Ascaris lumbricoides (13,3%). A prevalência de protozoários (42,6%) foi maior que de helmintos (31,1%). Para a maioria dos grupos analisados não houve diferença entre o quadro clínico de diarreia e o tipo e número de enteroparasita. O poliparasitismo foi detectado em 12,8% das amostras e o monoparasitismo em 46,5%. Os grupos etários de menor idade apresentaram predomínio de infecções por protozoários<br>Abstract: The parasitic infections are the major public health problems, presenting an endemic form in several areas of Brazil. They may present narrow relationship with social-demographical and environmental factors, such as: social-economical precarious conditions, consumption contaminated water, deficient nutritional condition of individuals and others, being frequently the infant population the most affected. In order to investigate the prevalence of intestinal parasites in indigenous and deprived populations and the key factors involved in the epidemiology of intestinal parasites, it was realized intestinal parasitological survey in residents of two Indian reservations: Bororó Reservation/MS and Xingu Reservation/MT (Kayabí and Juruna tribes); and also in residents of two cities: Pontes e Lacerda/MT and Ibateguara/Al. The data collection was conducted from 2002 to 2009. 2,754 faeces samples were analyzed by the methods of Faust, Hoffman, Kato-Katz, Rugai, Direct and modified Ziehl-Neelsen. The study obtained personal data and social-economical parameters. It was observed the presence of 73% of intestinal parasites in the Bororó Reservation, 62.77% in the Xingu Reservation, 52.61% in the cities of Pontes e Lacerda/MS and 67.42% in the Ibateguara city. The species of most prevalence in male individuals were Entamoeba coli (22.5%), Giardia duodenalis (11.6%), Entamoeba histolytica (13.9%) and Ascaris lumbricoides (13.6%). In female individuals were Entamoeba coli (24.1%), Giardia duodenalis (8.8%), Entamoeba histolytica/ E. díspar (13.8%) and Ascaris lumbricoides (13.3%). The prevalence of protozoan (42.6%) was higher than helminths (31.1%). For most analyzed groups there was no difference between the diarrhea clinical situation and the intestinal parasite type and number. The multiple intestinal parasite was detected in 12.8% of samples and monoparasitism in 46.5%. The minor age individuals presented the preponderance of protozoan infections<br>Doutorado<br>Parasitologia<br>Doutor em Parasitologia

Made available in DSpace on 2014-06-11T19:24:15Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-02-24Bitstream added on 2014-06-13T19:30:55Z : No. of bitstreams: 1 david_eb_me_botfm.pdf: 680578 bytes, checksum: 9ccd08f14d2896ddd592fa21e3bcaaba (MD5)<br>Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)<br>Universidade Estadual Paulista (UNESP)<br>O presente estudo foi realizado para investigar a prevalência de enteroparasitas em crianças e funcionários da creche do município de Pratânia, Estado de São Paulo, e caracterizar geneticamente os isolados de Giardia duodenalis obtidos dos indivíduos desse grupo empregando gdh e tpi como genes-alvo. Para isso, amostras de fezes de 141 crianças com idade de zero a seis anos e de 18 colaboradores foram colhidas e processadas pelo método de centrífugo-flutuação e pelo kit TF-test®. Nas 141 crianças, parasitas e/ou comensais intestinais foram diagnosticados em 64 (45,4%), das amostras, sendo Giardia (22%) e Cryptosporidium (22%) os parasitas mais freqüentes. Nos funcionários, Giardia foi diagnosticada em apenas uma amostra (5,5%) e o protozoário comensal Entamoeba coli (22,2%) foi o mais prevalente. Empregando técnicas baseadas em PCR, o DNA extraído de 86 (31 amostras positivas e 55 negativas para Giardia no exame microscópico) foi amplificado e os produtos obtidos foram seqüenciados. A análise de 35 seqüências revelou a ocorrência de infecção pelos genótipos A (28,6%) e B (71,4%), subtipos AI, AII, BIII/BSI e BIV, no entanto, associações entre os genótipos e fatores de risco para a infecção não foram observadas. O alinhamento das seqüências de gdh e tpi, revelou a presença de um a 12 SNPs (Single Nucleotide Polymorphisms) em isolados dos genótipos A e B, sendo que em alguns deles, as mutações foram responsáveis por alterações nas seqüências de aminoácidos. A infecção de seis crianças de um mesmo grupo e de uma funcionária por isolados classificados como genótipo B e apresentando seqüências nucleotídicas 100% idênticas entre si sugere a ocorrência de transmissão antroponótica na população estudada. Além disso, a maior prevalência do genótipo B, grupo predominante nas infecções humanas, reforça o fato de que a transmissão...<br>The present study was conducted to investigate the prevalence of intestinal parasites in children and employees of a daycare center of Pratania, São Paulo State, and to evaluate the genotypes Giardia duodenalis isolates obtained from individuals in this group by using gdh and tpi genes as targets. Fecal samples of 141 children aged zero to six years and 18 employees were processed by a flotation method and the TF-Test® kit. Of the 141 children samples, parasites and/or commensal organisms were diagnosed in 64 (45.4%) samples, and Giardia (22%) and Cryptosporidium (22%) were the most frequent parasites. In employees, Giardia was diagnosed in only one sample (5.5%) and the commensal protozoan Entamoeba coli (22.2%) was the most prevalent. DNA extracted from 86 samples (31 samples positive and 55 negative for Giardia by microscopic examination) was amplified by PCR and the products were sequenced. The analysis of 35 sequences revealed the occurrence of infection with genotypes A (28.6%) and B (71.4%) subtypes AI, AII, BIII / BIV and BSI, however, associations between genotypes and risk factors for infection were not observed. The alignment of the sequences revealed SNPs (Single Nucleotide Polymorphisms) in both genotypes A and B, altering the amino acid sequence of some isolates. Among isolates identified as genotype B, six obtained from children of the same group and one from the only one employee infected with Giardia showed gdh and tpi sequences different from reference ones, but 100% identical to each other. This data is suggestive of anthroponotic transmission in the studied population. In addition this finding, the higher prevalence of genotype B, the main group associated to human infections, reinforces the fact that anthroponotic transmission has importance in collective environments that meet children at an age that they still have not practice the basic precepts of hygiene.

One of the major constraints limiting the efficiency of sheep production is the control of gastrointestinal nematode parasites. Parasite control by strategic use of anthelmintics is threatened by the emergence of nematode populations that are resistant to the drugs available. It is therefore increasingly apparent that steps toward maintaining sustainable productivity in the growing face of anthelmintic resistance (AR) is required by farmers. To facilitate the uptake of sustainable approaches to parasite management, a comprehensive understanding of the various factors that may influence farmers’ decision-making processes is required. In order to establish which factors are influential, and determine their impact on farmers’ roundworm control behaviours, both qualitative and quantitative research methods were used including focus groups as well as attitudinal and behavioural questionnaires. A retrospective analysis was initially conducted of historical surveys designed to identify farm specific characteristics and parasite management practices including anthelmintic usage. The objectives of the analysis were to identify factors associated with uptake of best practice advice including farm characteristics and information sourcing. In addition, the implementation of sustainable roundworm control practices was investigated using two surveys conducted in 2000 and 2010. Logistic regression models were applied for univariable and multivariable analysis of dependent and independent variables. The next step was to conduct a series of focus groups in different geographic regions of Scotland. The main purposes were to explore sheep farmers’ attitudes towards different aspects of roundworm management, as well as to identify potential motivators and barriers to uptake of sustainable parasite control practices. The findings aided in the development of an attitudinal questionnaire used to canvass opinions representative on a national level. The concluding study involved a telephone survey of 400 Scottish sheep farmers, designed to elicit attitudes regarding roundworm control, AR and sustainable roundworm control practices. A quantitative statistical analysis technique (Structural Equation Modelling) was then used to test the relationships between socio-psychological factors and the uptake of sustainable roundworm control practices. The analysis of historical questionnaire data demonstrated evidence of a shift towards the use of practices to reduce the rate of AR development, most notably a decline in the practice of ‘dose and move’ as well as an overall reduction in treatment frequency. Statistical analysis identified significant associations between farm characteristics and specific treatment strategies. For example, larger farms were more likely to adopt a set treatment regimen (P=0.036), compared with smaller farms, which were more likely to treat based on clinical signs of infection (P=0.021). Sourcing of roundworm control information primarily from veterinarians was most associated with treatment timings with no parallels between time points. From the qualitative focus group studies conducted we identified four overarching themes impacting on sheep producers’ attitudes to roundworm control and best practice advice. These themes comprised: a lack of perceived need to change, the complexity of advice, the ease of implementation of recommended practices and the effectiveness of extension approaches. Additionally, the most important and implementable guidelines identified by sheep farmers were: ‘working out a roundworm control strategy with an advisor’ and ‘administering anthelmintics effectively’ with regard to correct drenching practice i.e. appropriate dosing, administration and drug storage procedures. These findings exhibited similarities with veterinarians’ rankings with also, ‘testing for AR’, ‘preserving susceptible parasites’ and ‘reducing dependence on anthelmintics’ receiving the lowest rankings for importance and implementability. The quantitative analysis from the attitudinal/behavioural questionnaire identified eleven factors with significant influences on the adoption of sustainable roundworm control practices. The key influences on overall adoption were farmers’ baseline understanding about roundworm control and self-reported confirmation of anthelmintic resistance in their flock. Additional positive influences included, positive attitudes to veterinary services, enterprise type and perceived risk of AR. Factors that were shown to have the greatest relative effects on individual parasite control practices included; the perceived resource requirements for implementing a quarantine strategy, farmers’ AR suspicions for instigating AR testing and the confirmation of AR for adopting faecal egg count monitoring. The findings have highlighted several factors which can influence sheep farmers’ decisions to reject or adopt recommended roundworm control practices. It is evident that the perceived complexity, lack of need and practicality of the current recommendations necessitates changes to how extension is designed and disseminated to farmers. The findings also suggest that improving farmers’ acceptance and uptake of diagnostic testing and improving underlying knowledge and awareness about nematode control is a significant target to influencing adoption of best practice behaviours. The importance of veterinarians as highly-trusted information resources validates the need for further engagement with veterinarians concerning sustainable parasite control approaches, to facilitate collaboration with farmers. The need for interaction between farmers and their advisors is key to resolving the issues raised to enable the necessary explanation, justification and execution of recommended practices to suit farmers’ needs and farming conditions.

Made available in DSpace on 2014-06-11T19:27:55Z (GMT). No. of bitstreams: 0 Previous issue date: 2005-06-24Bitstream added on 2014-06-13T18:56:48Z : No. of bitstreams: 1 mine_jc_me_arafcf.pdf: 726384 bytes, checksum: 6f4cf343dd34895158b4fd037741c3af (MD5)<br>Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)<br>Universidade Estadual Paulista (UNESP)<br>Blastocystis hominis é protozoário causador da infecção intestinal denominada blastocistose humana, cujo diagnóstico é realizado pelo exame coproparasitológico e por meio de técnicas de colorações permanentes que foram utilizadas neste estudo para avaliar a prevalência de Blastocystis hominis nos espécimes fecais de habitantes na região de Araraquara-SP. Foram estudadas 503 amostras de fezes submetidas às técnicas de exame direto a fresco, de Faust e cols., de Lutz e de Rugai, Mattos e Brisola, além das colorações pela hematoxilina férrica, tricrômio e de Kinyoun modificada. Do total das amostras analisadas 174 (34,6%) apresentaram-se positivas para a presença de parasitas intestinais. O protozoário e helminto mais freqüentes foram respectivamente: Entamoeba coli (14,6%) e Strongyloides stercoralis (6,7%). Blastocystis hominis foi observado em 23 (4,6%) amostras fecais com consistência predominantemente pastosa, não caracterizando quadro diarréico. Apesar da baixa prevalência de Blastocystis hominis encontrada na região de Araraquara, comparativamente a outras regiões brasileiras, é importante a realização do diagnóstico laboratorial desse protozoário. O encontro de Blastocystis hominis em material fecal é indicativo de contaminação de alimentos e água de consumo, desde que se admita a rota de transmissão oral-fecal deste parasita, o que implica na orientação da população sobre as medidas de saneamento básico e higiene como meio para se controlar problemas de saúde ocasionados pelos enteroparasitas.<br>Blastocystis hominis is a protozoan which causes an intestinal infection called human blasticistosis. Its diganosis is perfomed by stool examination and permanent staining techniques. Such methodologies were carried out on the present study in order to evaluate the prevalence of Blastocystis hominis in faecal specimens from the Araraquara region inhabitants. A total of 503 faecal samples were evaluated by the following techniques: examination fo fresh specimens, Lutz, Faust et al. and Rugai et al. besides the iron hemotoxylin, trichrome and modified Kinyon staining. Out of 503 stool samples examined 174 (34,6) were found to be positive for intestinal parasites. The most prevalent protozoan and helminth parasites were Entamoeba coli (14,6%) and Strongyloides stercoralis (6,7%) respectively. Balstocystis hominis was present in 23 (4,6%) stool samples, most of all of soft consistence and without diarrheic reports. Blastocystis hominis laboratorial diagnosis is important althought its prevalence has been low in Araraquara region. Blastocystis hominis findings is faecal specimens indicates the food and water contamination and since the transmission of this parasite is iral-faecal it implies that the population needs orientation about hygiene and basic sanitation conditions in order to control health problems caused by enteroparasites.

Resumo: O presente estudo foi realizado para investigar a prevalência de enteroparasitas em crianças e funcionários da creche do município de Pratânia, Estado de São Paulo, e caracterizar geneticamente os isolados de Giardia duodenalis obtidos dos indivíduos desse grupo empregando gdh e tpi como genes-alvo. Para isso, amostras de fezes de 141 crianças com idade de zero a seis anos e de 18 colaboradores foram colhidas e processadas pelo método de centrífugo-flutuação e pelo kit TF-test®. Nas 141 crianças, parasitas e/ou comensais intestinais foram diagnosticados em 64 (45,4%), das amostras, sendo Giardia (22%) e Cryptosporidium (22%) os parasitas mais freqüentes. Nos funcionários, Giardia foi diagnosticada em apenas uma amostra (5,5%) e o protozoário comensal Entamoeba coli (22,2%) foi o mais prevalente. Empregando técnicas baseadas em PCR, o DNA extraído de 86 (31 amostras positivas e 55 negativas para Giardia no exame microscópico) foi amplificado e os produtos obtidos foram seqüenciados. A análise de 35 seqüências revelou a ocorrência de infecção pelos genótipos A (28,6%) e B (71,4%), subtipos AI, AII, BIII/BSI e BIV, no entanto, associações entre os genótipos e fatores de risco para a infecção não foram observadas. O alinhamento das seqüências de gdh e tpi, revelou a presença de um a 12 SNPs (Single Nucleotide Polymorphisms) em isolados dos genótipos A e B, sendo que em alguns deles, as mutações foram responsáveis por alterações nas seqüências de aminoácidos. A infecção de seis crianças de um mesmo grupo e de uma funcionária por isolados classificados como genótipo B e apresentando seqüências nucleotídicas 100% idênticas entre si sugere a ocorrência de transmissão antroponótica na população estudada. Além disso, a maior prevalência do genótipo B, grupo predominante nas infecções humanas, reforça o fato de que a transmissão... (Resumo completo, clicar acesso eletrônico abaixo)<br>Abstract: The present study was conducted to investigate the prevalence of intestinal parasites in children and employees of a daycare center of Pratania, São Paulo State, and to evaluate the genotypes Giardia duodenalis isolates obtained from individuals in this group by using gdh and tpi genes as targets. Fecal samples of 141 children aged zero to six years and 18 employees were processed by a flotation method and the TF-Test® kit. Of the 141 children samples, parasites and/or commensal organisms were diagnosed in 64 (45.4%) samples, and Giardia (22%) and Cryptosporidium (22%) were the most frequent parasites. In employees, Giardia was diagnosed in only one sample (5.5%) and the commensal protozoan Entamoeba coli (22.2%) was the most prevalent. DNA extracted from 86 samples (31 samples positive and 55 negative for Giardia by microscopic examination) was amplified by PCR and the products were sequenced. The analysis of 35 sequences revealed the occurrence of infection with genotypes A (28.6%) and B (71.4%) subtypes AI, AII, BIII / BIV and BSI, however, associations between genotypes and risk factors for infection were not observed. The alignment of the sequences revealed SNPs (Single Nucleotide Polymorphisms) in both genotypes A and B, altering the amino acid sequence of some isolates. Among isolates identified as genotype B, six obtained from children of the same group and one from the only one employee infected with Giardia showed gdh and tpi sequences different from reference ones, but 100% identical to each other. This data is suggestive of anthroponotic transmission in the studied population. In addition this finding, the higher prevalence of genotype B, the main group associated to human infections, reinforces the fact that anthroponotic transmission has importance in collective environments that meet children at an age that they still have not practice the basic precepts of hygiene.<br>Orientador: Semíramis Guimarães Ferraz Viana<br>Coorientador: Teresa Cristina G. de Oliveira-Sequeira<br>Banca: Paulo Eduardo Martins Ribolla<br>Banca: Marcelo Urbano Ferreira<br>Mestre

Orientador: Marlene Tiduko Ueta<br>Tese ( doutorado) - Universidade Estadual de Campinas, Instituto de Biologia<br>Made available in DSpace on 2018-08-17T12:24:47Z (GMT). No. of bitstreams: 1 Rondon_MichelleVivianeSadosSantos_D.pdf: 5998356 bytes, checksum: 92f7ee4c29f1458fa4d49e970a955c26 (MD5) Previous issue date: 2010<br>Resumo: Os parasitas ocorrem praticamente em todos os níveis tróficos e sua transmissão pode depender da presença de uma variedade de hospedeiros intermediários, paratênicos e definitivos dentro do ecossistema. Exercem importantes efeitos sobre as populações de seus hospedeiros, alterando o comportamento, sucesso reprodutivo e a mortalidade. Por esse motivo, alguns autores os consideram importantes indicadores ambientais. O presente trabalho teve como objetivo estudar a epidemiologia e a biodiversidade de parasitas intestinais em mamíferos silvestres do reservatório do Jaguari, situado na cidade de Vargem/SP e do Parque Ecológico Prof. Hermógenes de Freitas Leitão Filho em Campinas/SP, e relacionar com os hábitos dos hospedeiros. Os espécimes de mamíferos foram capturados por armadilhas, posteriormente foram medidos, pesados, marcados, e após a coleta de fezes, os animais foram soltos. Para a pesquisa dos parasitas intestinais utilizaram-se os métodos de sedimentação, flutuação, e necrópsias foram realizadas em alguns casos. No reservatório de Jaguari foram realizadas 23 coletas entre os meses de agosto de 2005 a agosto de 2007. Os pequenos mamíferos capturados (N= 235) foram: Akodon montensis (71,5%), Calomys sp. (6,8%), Oligoryzomys nigripes (14%), Didelphis aurita (3,4%), Gracilinanus sp. (0,4%), Lutreolina crassicaudata (0,4%), Monodelphis sp. (2,1%) e Sylvilagus brasiliensis (1,3%). Também fora recolhidas amostras fecais (N=44), encontradas nas margens do reservatório de: Hydrochoerus hydrochaeris (56,8%), Lontra longicaudis (38,6%) e Puma concolor (4,5%). Das 303 amostras, 205 apresentaram-se positivas para parasitas, representando 67,6%. Foram encontrados: adultos de Cruzia tentaculata (0,5%), larvas de Nematoda (18,5%), ovos de Ascarididae (2,9%), Cruzia tentaculata (1,5%), Oxyuridae (0,9%), semelhantes à Dioctophyma sp. (0,5%), Syphacia sp. (0,5%), Toxocaridae sp. (0,5%), Trichostrongylidae (79%), Trichuridae (17%), Hymenolepis diminuta (0,9%), Hymenolepis nana (5,4%), Pseudophyllidea (0,9%), Taeniidae (0,5%), Trematoda (6,3%) e Acanthocephala (3,4%). Cistos de Amoebidae (1,9%), Giardia sp. (0,9%), semelhantes à Balantidium sp. (0,5%), oocistos de Coccidiida (10,2%), Eimeria sp. (0,9%) e trofozoitos de Amoebidae (2,4%), também foram encontrados. No Parque Ecológico, foram realizadas 16 coletas entre os meses de novembro de 2006 a fevereiro de 2008. Os pequenos mamíferos capturados (N= 103) correspondem aos roedores Myocastor coypus (6,8%), Nectomys squamipes (1,0%), Rattus rattus (20,4%), e o marsupial Didelphis albiventris (71,8%). O total de amostras fecais coletadas foi de 279, e 207 apresentaram-se positivas para parasitas, representando 74,2%. Foram encontrados adultos de Cruzia tentaculata (1,4%), larvas de Nematoda (24,1%), ovos de Ascarididae (3,4%), Capillaridae (2,9%), Cruzia tentaculata (67,6%), Oxyuridae (3,4%), semelhantes à Dioctophyma sp. (1,0%); semelhante à Syngamus sp. (6,3%), Spiruroidea (1,0%), Trichostrongylidae (21,2%), Trichuridae (19,8%), Hymenolepis diminuta (0,5%), Trematoda (8,2%), e Acanthocephala (1,9%). Oocistos de Coccidiida (39,6%), Eimeria sp. (4,3%) e Isospora sp. (1,0%) também estiveram presentes. Akodon montensis foi o animal mais frequente e com maior número de morfotipos de parasitas do reservatório de Jaguari, o mesmo ocorreu com o marsupial Didelphis albiventris no Parque Ecológico. Os parasitas mais frequentes foram os de ciclo monoxênico, que estão intimamente ligados aos hábitos dos animais estudados.<br>Abstract: The parasites occur practically in all trophic levels and their transmission can depend by the presence of a variety of intermediate, parathenic and definitive hosts within the ecosystem. They have important effects over their host populations as, behaviour changing, reproductive success and mortality. By these reasons, some authors consider then important environmental indicators. The objective of this studying was the epidemiology and the intestinals parasites biodiversity in wild mammals from the reservoir of Jaguari, located at the City of Vargem, São Paulo state, and relates with the hosts habits. The specimens were captured by traps, than measured, checked the weight, marked, the faeces were collected and the animals were released. For the intestinals parasites research, the sedimentation and fluctuation methods were used, and autopsies were performed in some cases. In the Jaguari reservoir 23 collects were performed between august 2005 to august 2007. The smalls captured mammals (N= 235) were: Akodon montensis (71.5%), Calomys sp. (6.8%), Oligoryzomys nigripes (14%), Didelphis aurita (3.4%), Gracilinanus sp. (0.4%), Lutreolina crassicaudata (0.4%), Monodelphis sp. (2.1%) and Sylvilagus brasiliensis (1.3%). Also faeces samples were collected (N=44) from the margins of the reservoir as: Hydrochoerus hydrochaeris (56.8%), Lontra longicaudis (38.6%) and Puma concolor (4.5%). From the 303 samples, 205 showed positive for parasites, representing 67.6%. Were found: adults of Cruzia tentaculata (0.5%), larvae of Nematoda (18.5%), Ascarididae eggs (2.9%), Cruzia tentaculata (1.5%), Oxyuridae (0.9%), similars to Dioctophyma sp. (0.5%), Syphacia sp. (0.5%), Toxocaridae sp. (0.5%), Trichostrongylidae (79%), Trichuridae (17%), Hymenolepis diminuta (0.9%), Hymenolepis nana (5.4%), Pseudophyllidea (0.9%), Taeniidae (0.5%), Trematoda (6.3%) and Acanthocephala (3.4%). Cysts of Amoebidae (1.9%), Giardia sp. (0.9%), similars to Balantidium sp. (0.5%), oocysts of Coccidiida (10.2%), Eimeria sp. (0.9%) and trophozoites of Amoebidae (2.4%), also were found. In the Ecological Park, were done 16 collects between November 2006 to February 2008. The small mammals captured (N= 103) were the rodents Myocastor coypus (6.8%), Nectomys squamipes (1.0%), Rattus rattus (20.4%), and the marsupial Didelphis albiventris (71.8%). The total faeces samples collected was 279, and 207 showed positive for parasites, representing 74.2%. Were found adults of Cruzia tentaculata (1.4%), Nematoda larvae (24.1%), Ascarididae eggs (3.4%), Capillaridae (2.9%), Cruzia tentaculata (67.6%), Oxyuridae (3.4%), similars to Dioctophyma sp. (1.0%); similar to Syngamus sp. (6.3%), Spiruroidea (1.0%), Trichostrongylidae (21.2%), Trichuridae (19.8%), Hymenolepis diminuta (0.5%), Trematoda (8.2%), and Acanthocephala (1.9%). Oocysts of Coccidiida (39.6%), Eimeria sp. (4.3%) and Isospora sp. (1.0%) were present as well. Akodon montensis was the most frequent animal and with the highest number of parasites morphotypes from the Jaguari reservoir, the same happened with the marsupial Didelphis albiventris in the Ecological Park. The most frequent parasites were those which have monoxenic cycle, which are intimately connected to the feed habits from the studied animals.<br>Doutorado<br>Parasitologia<br>Doutor em Parasitologia

Includes copies of other papers co-authored by the author at end of thesis. Includes bibliographical references (leaves 241-265) A rapid sampling technique has been adapted and used to measure nucleoside transport in a human-derived isolate of the intestinal protozoan parasite Giardia intestinalis (syn. G. lamblia)

Abstract Parasites are an inseparable part of the life of wild birds. They may cause morbidity, mortality or reduction in fecundity. Parasite distribution in hosts is typically not uniform and many host factors (e.g. age) may affect the pattern of distribution. Under certain conditions, parasites even have the potential to regulate the host population. The grouse species of Finnish forests — the capercaillie Tetrao urogallus , the black grouse Lyrurus tetrix and the hazel grouse Tetrastes bonasia — harbour several species of intestinal helminth parasites. The populations have fluctuated in cyclic manner but the mechanisms behind the cycles are largely unknown. I studied the interactions of forest grouse and their intestinal helminth parasites by using intestinal samples collected by hunters in five game management districts during eight years (1995–2002). The most common parasite species in the samples was the nematode Ascaridia compar. Also, three species of cestodes (Skrjabinia cesticillus, Paroniella urogalli and Hymenolepis sp.) were found. Large size, male gender and age over 1 year were connected with an increased probability and intensity of A. compar infection. Juvenile grouse were commonly infected with cestodes while in adults infections were quite rare. The influence of inbreeding on the susceptibility to parasite infections was studied in the capercaillie by analysing microsatellite heterozygosity. The less heterozygous birds were more likely to be infected with A. compar and were more intensely infected suggesting negative influence of inbreeding on parasite resistance. An indirect negative effect of parasites was found by comparing bags hunted with a trained dog or without a dog. Grouse infected by cestodes were significantly more common in the dog-assisted bag. Thus, cestode infection seemed to make grouse more vulnerable to canine predation. The interaction between grouse population dynamics and parasites was studied by analyzing the grouse densities obtained from annual wildlife counts and parasite indices. A. compar was most common and most abundant in the years of grouse population decline. The grouse population growth rate was negatively correlated with the annual mean abundance of A. compar. Relative survival but not breeding success decreased as the abundance of A. compar increased. The findings suggest that A. compar influences the dynamics of Finnish grouse even though regular cyclic dynamics are no longer evident<br>Tiivistelmä Loiset kuuluvat erottamattomana osana luonnonvaraisten lintujen elämään. Ne voivat aiheuttaa sairautta, kuolleisuutta tai hedelmällisyyden alentumista. Tyypillisesti loiset ovat levinneet isäntäpopulaatioon epätasaisesti ja monet isännän ominaisuudet (esim. ikä) vaikuttavat levinneisyyteen. Tietyissä oloissa loiset voivat jopa säädellä isäntäpopulaatiotaan. Suomalaiset metsäkanalinnut — metso Tetrao urogallus, teeri Lyrurus tetrix ja pyy Tetrastes bonasia — ovat useiden suolistoloismatolajien isäntiä. Metsäkanapopulaatiot ovat vaihdelleet syklisesti, mutta syklejä aiheuttavat mekanismit ovat yhä tuntemattomia. Tutkin metsäkanalintujen ja niiden suolistoloisten välisiä vuorovaikutuksia käyttäen metsästäjien vuosina 1995–2002 viidestä eri riistanhoitopiiristä keräämiä suolistonäytteitä. Yleisin loislaji näytteissä oli kanalintusuolinkainen, Ascaridia compar. Myös kolme heisimatolajia (Skrjabinia cesticillus, Paroniella urogalli ja Hymenolepis sp.) todettiin. Suuri koko, koirassukupuoli ja yli yhden vuoden ikä olivat yhteydessä suurempaan kanalintusuolinkaistartunnan todennäköisyyteen ja voimakkuuteen. Nuorilla (alle 1 v.) linnuilla heisimadot olivat yleisiä, kun taas aikuisilla tartunnat olivat varsin harvinaisia. Sisäsiittoisuuden vaikutusta loistartuntaherkkyyteen tutkittiin metsolla mikrosatelliittiheterotsygotian perusteella. Vähemmän heterotsygoottiset metsot olivat todennäköisemmin ja voimakkaammin suolinkaisten infektoimia, mikä viittaa sisäsiittoisuuden negatiiviseen vaikutukseen loisten vastustuskykyyn. Loisten epäsuora haitallinen vaikutus havaittiin, kun verrattiin koiran kanssa ja ilman koiraa metsästettyä lintusaalista. Heisimadot olivat selvästi yleisempiä linnuilla, jotka oli metsästetty koiran kanssa kuin ilman koiraa metsästetyillä. Heisimatotartunta näytti siis altistavan metsäkanoja koiraeläinten saalistukselle. Metsäkanalintu- ja loispopulaatioiden välistä vuorovaikutusta tutkittiin analysoimalla vuosittaisia metsäkanatiheyksiä ja loisten runsautta. Kanalintusuolinkainen oli yleisimmillään ja runsaimmillaan metsäkanatiheyden laskuvuosina. Metsäkanapopulaation vuosittainen kasvuvauhti korreloi negatiivisesti kanalintusuolinkaisen vuosittaisen runsauden kanssa. Suhteellinen elossasäilyvyys laski kanalintusuolinkaisen runsauden lisääntyessä, mutta lisääntymistuloksen suhteen ei ollut samaa ilmiötä. Löydökset viittaavat siihen, että kanalintusuolinkaisella on vaikutusta suomalaisten metsäkanalintukantojen vaihteluihin, vaikka syklisiä kannanvaihteluja ei enää havaitakaan

Foi estudado o perfil nosológico da população ribeirinha do Baixo Rio Machado em Rondônia, área de Amazônia Ocidental no Brasil. Neste estudo, escolhemos abordar o perfil das seguintes doenças: malária, hepatite viral B e C, parasitose intestinal e papilomavírus humano. Estas doenças foram priorizadas devido à importância clínica e epidemiológica e também escassez de estudos referentes nas populações ribeirinhas amazônicas. Para isto, foi realizado um estudo descritivo no Baixo Rio Machado que se localiza a 250 km de Porto Velho (capital do Estado de Rondônia) pela margem direita do Rio Madeira, que é um afluente volumoso do Rio Amazonas, no Brasil. Nesta área, vivem aproximadamente 806 pessoas distribuídas em 55 comunidades ribeirinhas isoladas. Em Rondônia, encontramos locais, como o Rio Machado, que abrigam a presença de portadores assintomáticos do Plasmodium, fato que pode contribuir para a persistência desta doença na região. Para caracterizar o perfil da malária ribeirinha, focalizando aspectos da infecção assintomática e sintomática, foram realizados hemoscopia e Reação em Cadeia da Polimerase (PCR) com amplificação do DNA ribossomal do parasita para Plasmodium vivax e Plasmodium falciparum para o diagnóstico da malária. Foi considerado como assintomático o paciente com hemoscopia positiva ou PCR positivo que permaneceu sem sintomas durante pelo menos 60 dias. Foi realizado acompanhamento longitudinal de 70 dias após o primeiro corte transversal (nC1=585 pessoas) para observação dos pacientes assintomáticos diagnosticados por hemoscopia, onde 25 pacientes (4,25%) permaneceram assintomáticos durante todo esse seguimento. Após, foi realizado mais dois cortes transversais a cada 6 meses (nC2=583 pessoas, nC3=607 pessoas), com tratamento dos indivíduos assintomáticos diagnosticados por PCR no corte transversal anterior ou hemoscopia atual e também dos casos sintomáticos. Foi estudado, na população que permaneceu na área durante toda a pesquisa (n=379), o perfil e o impacto do tratamento da infecção assintomática. Houve diminuição da infecção por P. falciparum de seis vezes e aumento da prevalência de malária por P. vivax de aproximadamente três vezes. O diagnóstico por PCR foi de 2-5 vezes mais eficiente do que por hemoscopia e a prevalência de infecção assintomática foi de 16 a 22% nos cortes transversais. Assim, foi observada a importância do tratamento dos indivíduos assintomáticos para o controle da malária no local, sendo que o incremento de malária por Plasmodium vivax deve ter ocorrido devido a recaídas da doença. Com o estudo entomológico da área, foi verificado que o vetor Anopheles possui atividade hematofágica moderada e principalmente em região peridomiciliar, sendo o A. darlingi a principal espécie da região. Existem poucos dados sobre a prevalência de hepatites na população ribeirinha de Rondônia, o que dificulta a vigilância epidemiológica nestas regiões amazônicas. Foi realizado um estudo de prevalência de hepatite B e C na área, com 123 pacientes pertencentes a 5 comunidades ribeirinhas, incluindo todas as pessoas que estavam presentes na área no momento do corte transversal, de todas as faixas etárias. Foi realizada sorologia para os marcadores de hepatite B: HbsAg, Anti-Hbc (total) e Anti-Hbs e sorologia para hepatite C com o marcador anti-HCV. Foram encontrados 12 (14,7%) pacientes com hepatite B aguda, 29 (38,7%) pacientes apresentaram imunidade vacinal para hepatite B e 7 (9,3%) pacientes apresentaram hepatite C, sendo que um (1,3%) paciente tinha co-infecção para hepatite B e C. Notou-se que esta população ribeirinha está exposta às hepatites virais, sendo necessário intensificar a vigilância epidemiológica na área, assim como a cobertura vacinal e também fornecer cuidados preventivos, curativos e paliativos em relação a estas doenças. A infecção pelo papilomavírus humano (HPV) é altamente prevalente, sendo detectada em aproximadamente 10% a 20% da população sexualmente ativa entre 15 e 49 anos de idade. A introdução de testes mais acurados para a detecção do DNA do HPV em investigações epidemiológicas confirmou a importância do HPV, principalmente dos tipos de alto risco, como o principal fator de risco para o desenvolvimento de neoplasia intra-epitelial cervical e câncer do colo uterino. Não foram encontrados estudos sobre a existência do HPV em populações ribeirinhas amazônicas na literatura médica. Foi realizado, então, um estudo transversal para levantar a prevalência de HPV nesta população, com 84 participantes em idade sexualmente ativa presentes na área. Após coleta de consentimento informado e questionário clínico-epidemiológico, foi realizado exame para isolar o HPV. A tipagem do HPV foi realizada com amplificação dos DNAs por PCR empregando iniciadores genéricos seguida de hibridização em pontos, capaz de identificar mais de 40 tipos diferentes de HPVs. Foram encontradas 18 pacientes contaminadas pelo HPV, perfazendo 21,4% da amostra. Os tipos de HPV encontrados foram: 53, 58, 31, 56, 16, 83, 55, 66, 45, 51, 40, 42, 6, 68. Os tipos de HPV mais freqüentes foram 51 (23%), 58 (19%), 53 (7,7%), 83 (7,7%), sendo HPV 16 encontrado em 3,8% das pacientes HPV positivas e a prevalência do HPV de alto risco oncogênco foi de 13,1%. Desta forma, foi encontrado alta prevalência de HPV na população ribeirinha amazônica estudada, evidenciando a necessidade de vigilância epidemiológica para câncer de colo uterino na região. Por fim, foi estabelecido a prevalência de parasitoses intestinais na população, abordando a correlação das enteroparasitoses com malária assintomática e anemia, realizando um inquérito coproparasitológico na área incluindo pacientes de todas as faixas etárias, que estivessem dispostos a participar do estudo. A análise das amostras foi realizada pelo método da Sedimentação Espontânea (método de Lutz ou Hoffmann, Pons & Janer). Entre os 268 exames de fezes realizados, a prevalência de parasitose intestinal encontrada na região em estudo foi de 86,6%. Entre os helmintos, Ascaris lumbricoides (47%), Ancilostomideos (37,3%), Trichuris trichiura (3,4%), Capillaria hepatica (2,3%), foram os parasitas mais encontrados. Entre os protozoários foram encontrados: Entamoeba coli (21,4%), Entamoeba histolytica (12%), Giardia intestinalis (15,4%), Endolimax nana (10,1%) e Iodamoeba butschlii (7,5%). A alta prevalência de parasitose intestinal encontrada na pesquisa foi concordante com outros dados desenvolvidos na Região Amazônica. Com este estudo, a região da Amazônia Ocidental insere-se no cenário geográfico mundial de distribuição da Capillaria hepatica, particularmente, a região do Baixo Rio Machado.<br>Was studied the nosological profile of the riverside population of the Baixo Rio Machado in Rondônia, Western Amazon area of Brazil. In this study, was chosed the profile of following diseases: malaria, hepatitis B and C, intestinal parasite and human papillomavirus. These diseases have been prioritized due to clinical and epidemiological importance and scarcity of studies in the riverside Amazon population. For this, was conducted a descriptive study in the Baixo Rio Machado which is located 250 km from Porto Velho (capital of Rondônia State) in the right bank of the Rio Madeira which is a large tributary of the Amazon River in Brazil. In this area, live about 806 people distributed in 55 riversides communities isolated. In Rondônia, we find places such as the Rio Machado, which shelter to the presence of asymptomatic carriers of Plasmodium, which may contribute to the persistence of this disease in the region. To characterize the profile of malaria riverside, focusing on aspects of symptomatic and asymptomatic infections, were performed haemoscopia and Polymerase Chain Reaction (PCR) with amplification of ribosomal DNA for the parasite Plasmodium vivax and Plasmodium falciparum to the diagnosis of malaria. Was considered as asymptomatic patient with positive haemoscopia or positive or PCR that remained without symptoms for at least 60 days. Longitudinal follow-up was conducted for 70 days after the first cross section (nC1 = 585 people) for observation of asymptomatic patients diagnosed by haemoscopia where 25 patients (4.25%) remained asymptomatic throughout the follow-up. After was performed two cross cuts every 6 months (nC2 = 583 people, nC3 = 607 people), with treatment of asymptomatic individuals diagnosed by PCR in previous transverse cross-sections or haemoscopia a current and also symptomatic cases. We studied in the population that remained in the area throughout the study (no= 379), the profile and impact of treatment of asymptomatic infection. There was a decrease in infection by P. falciparum of six times and increased prevalence of malaria by P. vivax approximately three times. The diagnosis by PCR was 2-5 times more efficient than haemoscopia and the prevalence of asymptomatic infection was 16 to 22% in transverse cross-sections. Thus, was saw the importance of treatment of asymptomatic individuals for the control of malaria in the place, and the increase in malaria by Plasmodium vivax should have occurred due to relapse of the disease. With entomological study of the area, was saw that the vector Anopheles has moderate blood activity and especially in peridomiciliary region and the A. darlingi the main species in the region. There are few data on the prevalence of hepatitis in riverside population of Rondônia, which makes surveillance in Amazonian regions. Was conducted a study of prevalence of hepatitis B and C in the area, with 123 patients belonging to 5 communities, including all the people who were present in the area at the cross section of all ages. Serology was performed for markers of hepatitis B: HBsAg, anti-HBc (total) and anti-HBs and serology for hepatitis C with anti-HCV marker. Was found 12 (14.7%) patients with acute hepatitis B, 29(38.7%) patients demonstrated immunity to hepatitis B vaccine and 7(9.3%) patients had hepatitis C, with one (1.3%) patient had co-infection for hepatitis B and C. Realize that this riverside population is exposed to viral hepatitis, being necessary to intensify epidemiological surveillance in the area, as well as immunization coverage and provide preventive care, curative and palliative for these diseases. The infection by human papillomavirus (HPV) is highly prevalent, being detected in approximately 10% to 20% of the sexually active population between 15 and 49 years of age. The introduction of more accurate tests for the detection of HPV DNA in epidemiological investigations confirmed the importance of HPV, particularly types of high risk, as the main risk factor for the development of cervical intraepithelial neoplasia and cervical cancer. Was found no studies on the existence of HPV in riverside Amazonian populations in the medical literature. The transversal cross-section study was take up to raise the prevalence of HPV in this population, with 84 participants in sexually active age in the area. After collection of informed consent and clinical-epidemiological questionnaire, was conducted to isolate the HPV test. The typing of HPV was taking with the DNAs for PCR using generic primers followed by hybridization in points, able to identify over 40 different types of HPV. Was found 18 patients infected by HPV, comprising 21.4% of the sample. The HPV types found were: 53, 58, 31, 56, 16, 83, 55, 66, 45, 51, 40, 42, 6, 68. The most common HPV types were 51 (23%), 58 (19%), 53 (7.7%), 83 (7.7%) with HPV 16 found in 3.8% of HPV positive patients and prevalence the high risk HPV oncogenic was 13.1%. Thus, was find high prevalence of HPV in riverside Amazonian population studied, highlighting the need for surveillance for cancer of the cervix in the region. Finally, was established the prevalence of intestinal parasites in the population, deal the correlation of intestinal with asymptomatic malaria and anemia, taking the fecal examination conducting in the area including patients of all ages, who were willing to participate in the study. The analysis of samples was performed by the spontaneous sedimentation method (method of Lutz or Hoffmann, Pons & Janer). Among the 268 fecal examinations, the prevalence of intestinal parasite found in the region under study was 86.6%. Among helminths, Ascaris lumbricoides (47%), Ancilostomideos (37.3%), Trichuris trichiura (3.4%), Capillaria hepatica (2.3%) were found more parasites. Among the protozoa were: Entamoeba coli (21.4%), Entamoeba histolytica (12%), Giardia intestinalis (15.4%), Endolimax nana (10.1%) and Iodamoeba butschlii (7.5%).The high prevalence of intestinal parasite found in the survey was consistent with other data developed in the Amazon region. In this study, the region of the Western Amazon is part of the scenario of global geographic distribution of Capillaria hepatica, particularly the region of the Baixo Rio Machado.

[ES] La producción en acuicultura se ha visto menguada por aparición de enfermedades en los sistemas de cría de peces. En concreto, en la dorada (Sparus aurata), hay dos parásitos destacados: Enteromyxum leei (Myxozoa) y Enterospora nucleophila (Microsporidia). Hasta la fecha, para ninguno de los dos se ha establecido un cultivo in vitro, y solo para E. leei se ha conseguido establecer un modelo de mantenimiento de la infección in vivo. La presente tesis pretende incrementar el conocimiento sobre estos parásitos y sus relaciones con el hospedador, sentando las bases para generar soluciones que puedan ser aplicadas en la acuicultura. El objetivo con E. leei fue estudiar la inmunidad adquirida inducida en la dorada y la posibilidad de generar herramientas de diagnóstico y vacunas frente a esta enfermedad. Para ello, primero se demostró la resistencia del pez al parásito tras una segunda exposición, la cual duró hasta 16 meses. Además, la resistencia parece estar correlacionada con altos niveles de inmunoglobulina (Ig) M específica en sangre, y una alta expresión de Igs, incluso antes de la re-exposición al parásito. El siguiente paso fue afinar el protocolo de infección con E. leei. Los resultados mostraron que una semana es suficiente para transmitir la infección de E. leei por efluente, independientemente de la temperatura. Tras la demostración de la respuesta adaptativa eficaz frente a E. leei, y al disponer de un modelo de infección refinado, se realizó un ensayo de inmunización pasiva. Aquí, los resultados mostraron que los anticuerpos especi'ficos efectivamente consigue ralentizar la invasión del intestino por el parásito y disminuir los síntomas de la enfermedad. Paralelamente, el resultado del análisis del repertorio de las regiones variables de la IgM e IgT del intestino peces resistentes mostró la inducción de una respuesta policlonal en las ce'lulas B. En base a estos resultados, se realizó una búsqueda de antígenos de E. leei que pudieran ser utilizados como candidatos para la producción de vacunas (análisis proteómico) o herramientas de diagnóstico (análisis in silico). Para ello, se ensambló un transcriptoma de novo utilizando una muestra mixta de intestino de dorada y parásito. Los resultados dieron lugar a 7 y 12 candidatos en la búsqueda in silico y proteómica, respectivamente. En los estudios de E. nucleophila, debido a que fue descrita muy recientemente, el punto de partida fue más básico. Las muestras de este parásito solo se pueden obtener de brotes naturales en piscifactorias. Por ello, primero se realizó un estudio de caracterización de la patología de la infección a partir de peces infectados naturalmente. En etapas tempranas de la infección, el parásito se localiza principalmente en el intestino, pero meses después, la prevalencia en intestino baja e incrementa en los órganos hematopoyéticos y el esto'mago. Los signos clínicos de la infección consistieron en una reducción significativa del crecimiento, emaciación, y palidez de las paredes intestinales. A nivel celular, en los casos ma's graves se observó hipercelularidad en el epitelio intestinal y proliferación de ce'lulas rodlet, un elevado número de linfocitos en la base del epitelio e infiltración de granulocitos acidófilos en el epitelio intestinal. Finalmente se probaron varias formas de transmisión horizontal de E. nucleophila (cohabitación, efluente, intubación oral y anal) con para desarrollar un modelo de mantenimiento in vivo. Se consiguió la transmisión el parásito por todas las vías, pero con una disminución de prevalencia a lo largo del tiempo. Variables como la temperatura, la dosis, y el estado de los peces donantes parecen ser más determinantes que la ruta seleccionada para la transmisión. Entre las rutas probadas, la intubación anal parece ser la más prometedora, pero ninguna de ellas fue capaz de reproducir los signos clínicos observados en las infecciones naturales.<br>[CA] La producció en aqüicultura s'ha vist minvada per aparició de malalties en els sistemes de cria de peixos. En concret, en l'orada (Sparus aurata), hi ha dos paràsits destacats: Enteromyxum leei (Myxozoa) i Enterospora nucleophila (Microsporidia). Fins avui, per a cap dels dos s'ha establert un cultiu in vitro, i només per a E. leei s'ha aconseguit establir un model de manteniment de la infecció in vivo. La present tesi pretén incrementar el coneixement sobre aquests paràsits i les seves relacions amb l'hoste, establint les bases per a generar solucions que puguin ser aplicades en l'aqüicultura. L'objectiu amb E. leei va ser estudiar la immunitat adquirida induïda en l'orada i la possibilitat de generar eines de diagnòstic i vacunes enfront d'aquesta malaltia. Per a això, primer es va demostrar la resistència del peix al paràsit després d'una segona exposició, la qual va durar fins a 16 mesos. A més, la resistència sembla estar correlacionada amb alts nivells d'immunoglobulina (Ig) M específica en sang, i una alta expressió de Igs, fins i tot abans de la re-exposició al paràsit. El següent pas va ser afinar el protocol d'infecció amb E. leei. Els resultats van mostrar que una setmana és suficient per a transmetre la infecció de E. leei per efluent, independentment de la temperatura. Després de la demostració de la resposta adaptativa eficaç enfront de E. leei, i en disposar d'un model d'infecció refinat, es va realitzar un assaig d'immunització passiva. Aquí, els resultats van mostrar que els anticossos específics efectivament aconsegueix alentir la invasió de l'intestí pel paràsit i disminuir els símptomes de la malaltia. Paral·lelament, el resultat de l'anàlisi del repertori de les regions variables de la IgM i IgT de l'intestí peixos resistents va mostrar la inducció d'una resposta policlonal en les cèl·lules B. Sobre la base d'aquests resultats, es va realitzar una cerca d'antígens de E. leei que poguessin ser utilitzats com a candidats per a la producció de vacunes (anàlisis proteómico) o eines de diagnòstic (anàlisi in silico). Per a això, es va assemblar un transcriptoma de novo utilitzant una mostra mixta d'intestí d'orada i paràsit. Els resultats van donar lloc a 7 i 12 candidats en la cerca in silico i proteòmica, respectivament. En els estudis de E. nucleophila, pel fet que va ser descrita molt recentment, el punt de partida va ser més bàsic. Les mostres d'aquest paràsit només es poden obtenir de brots naturals en piscifactorias. Per això, primer es va realitzar un estudi de caracterització de la patologia de la infecció a partir de peixos infectats naturalment. En etapes primerenques de la infecció, el paràsit es localitza principalment en l'intestí, però mesos després, la prevalença en intestí baixa i incrementa en els òrgans hematopoètics i l'estómac. Els signes clínics de la infecció van consistir en una reducció significativa del creixement, emaciació, i pal·lidesa de les parets intestinals. A nivell cel·lular, en els casos més greus es va observar hipercelularidad en l'epiteli intestinal i proliferació de cèl·lules rodlet, un elevat nombre de limfòcits en la base de l'epiteli i infiltració de granulòcits acidòfils en l'epiteli intestinal. Finalment es van provar diverses formes de transmissió horitzontal de E. nucleophila (cohabitació, efluent, intubació oral i anal) amb per a desenvolupar un model de manteniment in vivo. Es va aconseguir la transmissió el paràsit per totes les vies, però amb una disminució de prevalença al llarg del temps. Variables com la temperatura, la dosi, i l'estat dels peixos donants semblen ser més determinants que la ruta seleccionada per a la transmissió. Entre les rutes provades, la intubació anal sembla ser la més prometedora, però cap d'elles va ser capaç de reproduir els signes clínics observats en les infeccions naturals.<br>[EN] Aquaculture production is hampered by the emergence of parasite diseases in fish farming systems. Among them, in Sparus aurata, there are two important enteric parasites described: Enteromyxum leei (Myxozoa) Enterospora nucleophila (Microsporidia). To date, no in vitro culture has been established for either parasite, and only for E. leei was it possible to establish a model for maintaining the infection in vivo. The aim of this thesis is to gain new knowledge about these parasites and their relationship with the host, also the basic foundations for generating solutions that can be applied in aquaculture. The general objective for E. leei was to study the acquired immunity induced in gilthead bream and the possibility of generating diagnostic tools and vaccines against this disease. To this end, resistance against the parasite was assessed with a second exposure against the parasite, which showed a resistance for at least 16 months. Besides resistance seemed to be correlated with high levels of specific immunoglobulin (Ig) M in blood, and a high expression of Igs, in particular, the soluble forms, even before re-exposure to the parasite. The next step was refining the protocol for effluent infection with E. leei by studying infection at different exposure time points, temperatures and population densities. The results showed that one week of exposure is sufficient to spread E. leei infection by effluent, regardless of temperature. After demonstrating the resistance against E. leei, and with a refined infection model, a passive immunization assay was performed. The results showed that the serum with specific antibodies effectively slows down the invasion of the gut by the parasite and reduces the symptoms of the disease. At the same time, the analysis of the repertoire of the variable regions of intestinal IgM and IgT showed an induction of a polyclonal response in B cells. On the basis of these results, a research was carried out for E. leei antigens that could have use as candidates for the production of vaccines (proteomic study) or diagnostic tools (in silico study) using the parasite transcriptomic data. To do this, a de novo transcriptome was assembled using a mixed sample of gilthead sea bream and parasite, with a posterior filtrate of the sequences. The In silico and proteomic analysis search resulted in 7 and 12 transcripts, respectively, which are being used for diagnostic and vaccine production. The starting point was more basic in E. nucleophila studies, since this is a recently described disease. The samples of this parasite can only be obtained from natural outbreaks in fish farms. Therefore, first study was carried out to characterize the pathology of the infection of naturally infected fish. In the early stages of the infection, the parasite is mainly located in the intestine, but months later, the prevalence is lower in the intestine and increases in the hematopoietic organs and the stomach. Clinical signs of infection were significant reduction in growth, wasting, and intestinal walls paleness. At the cellular level, in the most severe cases hypercellularity in the intestinal epithelium, proliferation of rodlet cells, high number of lymphocytes at the base of the epithelium and infiltration of acidophilic granulocytes in the intestinal epithelium were observed. Finally, horizontal transmission of E. nucleophila was tried using different transmission methods: cohabitation, effluent, and oral and anal intubation. Transmission of the parasite was achieved with all routes, but there was a decrease in prevalence over time in all cases except for the anal route. Variables such as temperature, dose, and the status of the donor fish appear to be more important than the selected route. Among the routes tested, anal intubation seemed to be the most promising, as it was sustained over a longer period of time, but none of them was able to reproduce the same clinical signs of infection observed in natural infections.<br>The authors kindly acknowledge the collaboration of anonymous fish farming companies allowing access to the animals during the disease outbreaks. We thank J. Monfort and L. Rodríguez (IATS-CSIC) for the technical assistance on histological processing.This work has been carried out with financial support from the European Union and the Spanish Ministry of Economy and Competitiveness (MINECO) under grant projects ParaFishControl (H2020-634429) and AGL2013-R-48560-C2-2-R, respectively. APS was contracted under ParaFishControl project. Primer sequences and access to the gilthead sea bream transcriptomic database were kindly provided by Prof. J. Pérez-Sánchez of the IATS- Nutrigenomics group. The authors thank I. Vicente for fish maintenance and technical assistance during samplings. The authors thank P. Boudinot (INRAE) for his help in designing and interpreting the immunoglobulin repertoire study and results, J. Pérez-Sánchez (IATS-CSIC) for providing access to the gilthead sea bream genome sequences to perform the repertoire analysis.This work was funded by the European Research Council (ERC Consolidator Grant 2016 725061 TEMUBLYM).<br>Picard Sánchez, MA. (2021). Control of enteric parasitic diseases of farmed gilthead sea bream: New insights into Enteromyxum leei (Myxozoa) and Enterospora nucleophila (Microsporidia) infections [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/167035<br>TESIS

Orientadores: Jancarlo Ferreira Gomes, Alexandre Xavier Falcão<br>Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia<br>Made available in DSpace on 2018-08-23T15:19:46Z (GMT). No. of bitstreams: 1 Carvalho_JulianaBarboza_M.pdf: 1818938 bytes, checksum: d44ab358d0482575e52d1f77a3f4c07b (MD5) Previous issue date: 2013<br>Resumo: As parasitoses intestinais são altamente prevalentes no mundo, estando entre as maiores causadoras de doenças e óbitos em seres humanos. Atualmente, o diagnóstico laboratorial destas parasitoses é realizado por meio de procedimentos técnicos manuais, desenvolvidos na sua grande maioria há décadas, o que justifica a aplicabilidade de técnicas mais sensíveis e práticas para esta finalidade, visando obter resultados eficientes, especialmente em programas governamentais direcionados à Saúde Pública. Sendo assim, o objetivo do projeto foi de avaliar e validar uma nova técnica parasitológica, denominada TF-Test Modified, em comparação com três técnicas parasitológicas convencionais consagradas pela literatura: TF-Test Conventional; Rugai, Mattos e Brisola; e Kato-Katz/Helm-Test. As etapas do trabalho consistiram em realizar coleta de material fecal de 457 indivíduos localizados em regiões endêmicas para parasitoses no município de Campinas, SP; no processamento laboratorial de 1.828 exames; no diagnóstico de 14 espécies parasitárias; e na análise estatística qualitativa de resultados de maneira abrangente. Dentre as espécies parasitárias encontradas, helmintos e protozoários intestinais foram detectados em 42,23% de indivíduos pela técnica de TF-Test Modified, ante 36,76% por TF-Test Conventional, 5,03% por Kato-Katz/Helm-Test, e 4,16% por Rugai, Mattos e Brisola. Destes casos, 54,40% de infecção simples dos indivíduos demonstrou serem portadores de monoparasitismo. A nova técnica parasitológica de TF-Test Modified, quando comparada com as demais técnicas, apresentou alto valor de infecção, como exemplo para dupla, tripla e múltipla, de maneira a perfazer um total de 98,37% de infecções. Ademais, a nova técnica apresentou índice Kappa com grau de concordância Quase Perfeito em todos os parâmetros avaliados com estimativa de 95% (P<0,05), permitiu encontrar com alta eficiência diagnóstica todas as espécies parasitárias estudadas, mostrou um notável diagnóstico verdadeiro, especialmente quando analisada comparativamente com as outras três técnicas convencionais. O atual estudo permitiu concluir que a técnica de TF-Test Modified pode ser utilizada de forma abrangente no diagnóstico qualitativo de protozoários e helmintos intestinais de humanos. O ganho de sensibilidade diagnóstica proporcionada por esta nova técnica deverá ser de estimável contribuição para o diagnóstico individual laboratorial, inquéritos populacionais e controle das parasitoses intestinais, de modo a repercutir em contribuição social<br>Abstract: Intestinal parasites are highly prevalent worldwide and is among the largest cause of illness and death in humans. Currently, the laboratory diagnosis of these parasites is accomplished through technical procedures manuals, developed mostly for decades, justifying the applicability of more sensitive techniques and practices for this purpose, to obtain effective results, especially in government programs aimed at Public Health. Thus, the objective of the project was to evaluate and validate a new technique parasite, called TF-Test Modified, compared with three conventional parasitological techniques enshrined in literature: TF-Test Conventional; Rugai, Mattos and Brisola, and Kato-Katz / Helm-Test. The steps of the work consisted of conducting a collection of fecal samples from 457 individuals located in regions endemic for parasitic infections in Campinas, SP, in laboratory processing of 1,828 examinations, the diagnosis of 14 parasitic species, and the qualitative statistical analysis of results so comprehensive. Among the species found parasitic, helminths and intestinal protozoa were detected in 42,23% of subjects using the technique of TF-Test Modified, against 36,76% by TF-Test Conventional, 5,03% by Kato-Katz/Helm-Test, and 4,16% Rugai, Mattos and Brisola. Of these cases, 54,40% of single infections of individuals were shown to be carriers of monoparasitism. The new technique parasitological TF-Test Modified compared to other techniques of infection showed a high value, for example double, triple and multiple so as to make a total of 98,37% infections . Moreover, the new technique presented Kappa index level of agreement with Almost Perfect in all parameters with estimated 95% (P <0.05), allowed to meet with high diagnostic efficiency all parasitic species studied showed remarkable true diagnosis, especially when viewed in comparison with other three conventional techniques. The current study showed that the technique TF-Test Modified can be used comprehensively in qualitative diagnosis of intestinal protozoa and helminths of humans. The gain in diagnostic sensitivity afforded by this new technique should be estimable contribution to the individual diagnostic laboratory, population surveys and control of intestinal parasites, in order to reflect on social contribution<br>Mestrado<br>Parasitologia<br>Mestra em Parasitologia

Submitted by Erika Demachki (erikademachki@gmail.com) on 2015-05-21T19:50:01Z No. of bitstreams: 2 Dissertação - Hanstter Hallison Alves Rezende - 2015.pdf: 1875029 bytes, checksum: a2cbf1f53df5830e4fdd38a866fc6ff4 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5)<br>Approved for entry into archive by Erika Demachki (erikademachki@gmail.com) on 2015-05-21T19:53:32Z (GMT) No. of bitstreams: 2 Dissertação - Hanstter Hallison Alves Rezende - 2015.pdf: 1875029 bytes, checksum: a2cbf1f53df5830e4fdd38a866fc6ff4 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5)<br>Made available in DSpace on 2015-05-21T19:53:32Z (GMT). No. of bitstreams: 2 Dissertação - Hanstter Hallison Alves Rezende - 2015.pdf: 1875029 bytes, checksum: a2cbf1f53df5830e4fdd38a866fc6ff4 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2015-02-20<br>Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG<br>The domestic cat is an important zoonotic agent for humans, especially as a transmissor of Toxoplasma gondii. The cat is capable of the elimination of millions of oocysts in the environment leading to a massive contamination of these places and leading to a public health issue. Toxoplamosis is a disease with high prevalence throughout the world and the cat is responsible for the parasite’s perpetuation. The evaluation of the accuracy of parasitological diagnostic methods is of vital importance to improve the veterinarian laboratory diagnosis through the use of high sensitivity, specificity and reproducible tests. The aim of this study was to verify the prevalence of intestinal parasites from stray cats in Goiania-GO, to verify the seroprevalence of anti-T. gondii antibodies through the modified aglutination test (MAT) and to perform the evaluation of the accuracy of the parasitological tests applied. Therefore 155 samples of feces and 50 blood samples from stray cats from the Zoonosis Center of Goiania were collected during the year of 2012. The feces samples were processed by the Willis (gold standard), Sheather, Faust and Hoffman-Janer-Pons-Lutz (HJPL) techniques. The MAT was performed in the blood samples. The accuracy was performed through the determination of sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and kappa index (k). The intestinal parasites prevalence in the feces from stray cats was 71.68% (115/154). The most prevalent helminth was Ancylostomatids (83.48%, 96/115) and the most prevalent protozoan in monoparasitism was Cystoisospora sp (7.83%, 9/115). T. gondii presented high prevalence (18.27%, 21/115) in fecal samples, 64% in blood samples (32/50) while 36% seronegative (18/50). The accuracy showed that the HJPL and Sheather techniques presented association to the golden standard for the intestinal parasites diagnosis. For the specific diagnosis of T. gondii oocysts the Faust techniques was the one with high accuracy in comparasion to the golden standard, with we emphasize the importance of using two parasitological techniques for the diagnosis of intestinal parasites<br>O gato doméstico é um importante transmissor de agentes zoonóticos para o homem, em especial Toxoplasma gondii. O gato infectado é capaz de eliminar milhões de oocistos no meio ambiente, provocando uma contaminação maciça nestes locais, sendo considerado um problema de saúde pública. A toxoplasmose é uma doença com alta prevalência em todo o mundo, sendo o gato responsável pela perpetuação do parasito. Avaliar a acurácia de métodos parasitológicos é de extrema importância para melhorar o diagnóstico laboratorial veterinário, utilizando testes de alta sensibilidade, especificidade e reprodutibilidade. O objetivo deste trabalho foi verificar a prevalência de parasitos intestinais em gatos errantes em GoiâniaGO, verificar a soroprevalência de anticorpos anti-T. gondii pelo teste de aglutinação modificado (MAT) e realizar a avaliação da acurácia de técnicas parasitológicas aplicadas no diagnóstico de parasitos intestinais. Foram coletadas 155 amostras de fezes no ano de 2012 no Centro de Controle de Zoonoses de Goiânia-GO e 50 amostras de sangue de gatos no mesmo período. As amostras de fezes foram processadas pelos métodos de Willis (padrão-ouro), Sheather, Faust e Hoffman-Janer-Pons-Lutz (HJPL). Nas amostras de sangue foi realizado o MAT. A análise de acurácia foi realizada determinando a sensibilidade, especificidade, valor preditivo positivo (VPP), valor preditivo negativo (VPN) e índice kappa (k). A prevalência de parasitos intestinais no ano de 2012 foi de 71,68% (115/154), sendo os Ancilostomídeos o helminto mais prevalente 83,48% (96/115), o protozoário mais prevalente foi Cystoisospora sp. em monoparasitismo 7,83% (9/115). T. gondii apresentou alta prevalência: 18,27% (21/115). Ao analisar a soroprevalência de um grupo de 50 animais, 64% (32/50) eram soropositivos e 36% (18/50) soronegativos. A análise de acurácia demonstrou que as técnicas de HJPL e Sheather apresentaram associação ao padrão-ouro para o diagnóstico de parasitos intestinais. No diagnóstico específico de oocistos de T. gondii a técnica de Faust apresentou melhor acurácia comparada ao padrão-ouro, com isso ressaltamos a importância do uso de duas técnicas parasitológicas para o diagnóstico de parasitos intestinais.

Made available in DSpace on 2014-12-17T14:16:27Z (GMT). No. of bitstreams: 1 MilenaTFA_DISSERT.pdf: 539795 bytes, checksum: f3249686901e2101c3e1aa253a03c1da (MD5) Previous issue date: 2009-07-31<br>Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior<br>The exclusive breastfeeding in the first six months of life practice is essential for the health and development of children. Studies show there is in human milk protective antibodies against intestinal parasites and a relationship between the absence of breastfeeding or weaning and parasitic infections. This work was a prospective cohort study involving 34 pregnant adolescents attended school in Maternity Januario Cicco / RN and their children, to assess the influence of breastfeeding on intestinal parasites in them. Thus, the parasitic infection was investigated by examination of feces parasitological and environmental factors by use of questionnaires. The average age of the mothers studied was 16.2 years. Of these 76.4% were infected and the most prevalent species of parasites were Entamoeba histolytica/dispar (76.9%), Giardia lamblia (19.2%) and Ascaris lumbricoides (11.5%). The infected children had an average age of 5.1 months and the frequency of parasites was 61.7%. The infection was earlier detecctada with 45 days of life. The most common parasites were Giardia lamblia (40%), Entamoeba histolytica/dispar (35%), Ascaris lumbricoides (5%) and Ancylostoma (5%). The average length of exclusive breastfeeding was 2.2 months. There was an association between increased duration of exclusive breastfeeding and increased time to detection of parasites in the feces of children. There was no statistically significant correlation between the socio-economic conditions and cultural and breastfeeding. The data reinforce the importance of breastfeeding to protect the children in its first year of life<br>A amamenta??o exclusiva nos primeiros seis meses de vida constitui pr?tica indispens?vel para a sa?de e o desenvolvimento da crian?a. Estudos mostram existir no leite humano anticorpos protetores contra enteroparasitos e uma rela??o entre a aus?ncia do aleitamento ou desmame e infec??es parasit?rias. Neste trabalho realizou-se um estudo de coorte prospectivo, envolvendo 34 gestantes adolescentes atendidas na Maternidade Escola Janu?rio Cicco/RN e seus filhos, para avaliar a influ?ncia do aleitamento materno nas parasitoses intestinais dos mesmos. Para tanto, a infec??o parasit?ria foi investigada por exame parasitol?gico de fezes e os fatores ambientais por aplica??o de question?rios. A idade m?dia das m?es estudadas variou de 14 a 19 anos, com m?dia de 16,2 anos. Destas, 76,4% estavam infectadas e as esp?cies de parasitos mais prevalentes foram Entamoeba histolytica/dispar (76,9%), Giardia lamblia (19,2%) e Ascaris lumbricoides (11,5%). Os filhos infectados apresentaram m?dia de idade de 5,1 meses e a frequ?ncia de parasitoses foi de 61,7%. A infec??o mais precoce foi detecctada com 45 dias de vida. Os parasitas mais freq?entes foram: Giardia lamblia (40%), Entamoeba histolytica/dispar (35%), Ascaris lumbricoides (5%) e Ancylostomo (5%). O tempo m?dio de amamenta??o exclusiva foi de 2,2 meses. Observou-se associa??o entre um maior tempo de amamenta??o exclusiva e um maior tempo para a detec??o de parasitos nas fezes das crian?as. N?o foi encontrada correla??o estatisticamente significativa entre as condi??es socioecon?micas e culturais e a amamenta??o. Os dados obtidos refor?am a grande import?ncia do aleitamento materno em proteger a crian?a no seu primeiro ano de vida

Orientador: Urara Kawazoe<br>Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia<br>Made available in DSpace on 2018-08-23T00:21:20Z (GMT). No. of bitstreams: 1 MonteiroFilho_LuizPauloCobra_M.pdf: 2118812 bytes, checksum: a948577e483cc6e6c98d3bb599dd8489 (MD5) Previous issue date: 2011<br>Resumo: Em jardins zoológicos as aves cativas silvestres e as exóticas podem estar expostas a algum tipo de estresse, o que causa imunossupressão em alguns indivíduos e, conseqüentemente, facilita o parasitismo até por parasitos pouco patogênicos. Tornam-se importantes, análises periódicas da relação parasito-hospedeiro em aves cativas para a percepção e entendimento da situação no controle de doenças, principalmente considerando a possibilidade da identificação de novas espécies de parasitos intestinais em novos hospedeiros, em especial os protozoários. Com esse objetivo, foram realizados levantamentos coproparasitológicos em três jardins zoológicos (Americana-SP, Guarulhos-SP e Alfenas-MG) e em um criadouro de aves silvestres e exóticas (Sítio da Ferradura em Poços de Caldas). Amostras frescas de fezes foram coletadas em solução de dicromato de potássio a 2,0 % e processadas usando o método de Hoffman, Pons & Janer modificado, seguido do Método de McMaster modificado (flutuação em solução saturada de NaCl). Os parasitos identificados com maior freqüência foram os coccídios Eimeria spp. e o nematoda Capillaria spp. Nos Zoológicos de Americana e Guarulhos, situados no Estado de São Paulo, a prevalência média de Capillaria spp. foi de 57,1 % e 66,7 % e de Eimeria spp. foi de 42,9 % e 33,3 %, respectivamente. No Jardim Zoológico de Alfenas (MG) observou-se apenas a presença de coccídios (100,0 %), sendo que se trata de um plantel com apenas 20 aves, do setor extra e dos recintos de exposição. No Criadouro de Poços de Caldas, identificou-se 36,0 % tanto para a Capillaria spp. como para a Eimeria spp. Considerando o total de amostras examinadas das quatro instituições, o gênero de ave que apresentou maior parasitismo por coccídio foi o Ramphastos spp., apesar da maioria das aves examinadas pertencerem a Ordem Psittaciformes. Pela análise das medidas de comprimento e largura dos oocistos encontrados em Rhamphastos toco (24,54 ?m ±1,17 x 23,83 ?m ±1,54) e em Coragyps atratus (20,99 ?m ±1,24 x 17,59 ±1,78 ?m e 17,92 ?m ±0,74 x 15,59 ?m ±0,96) com medidas, morfologia e hospedeiros distintos das espécies descritas na literatura, concluiu-se a identificação de três novas espécies de Eimeria no presente estudo. Além dessas espécies, três outras espécies de Eimeria (Eimeria forresteri, E. amazonae e E. ochrocephalae) descritas na literatura também foram identificadas. A partir da avaliação dos resultados obtidos no levantamento parasitológico, algumas medidas e procedimentos de manejo referentes à prevenção auxiliaram no controle da coccidiose e capilariose em ambientes com aves cativas<br>Abstract: Wild and exotic captive birds living in the Zoo are under stress conditions which could affect their immune system and acquire parasite infections even those with low pathogenicity. Periodical analysis of intestinal parasites, especially protozoans are important for their control and eventually identifications of new species in new hosts. The objective of this study was to identify intestinal parasite species in captive birds of Zoo Garden of Americana, São Paulo; Zoo Garden of Guarulhos, SP and Zoo Garden of Alfenas, Minas Gerais and the Wild and Exotic Birds Breeding Place located in the ?Sítio da Ferradura?, Poços de Caldas, MG. Fresh fecal samples were collected and placed in 2,0 % potassium dichromate solution and examined after using modified concentrated method of sedimentation (Hofman, Pons & Janer) and McMaster modified floatation method. The most frequent parasites identified were Eimeria spp. and Capillaria spp. In the Zoo Garden of Guarulhos and Americana, the prevalence of Capillaria spp. was 57,1 % and 66,7 % and of Eimeria spp. was 42,9 % and 33,3 % respectively. In the Zoo Garden of Alfenas, with a small number of 20 birds, only Eimeria spp. was identified (100 %) while in the Bird Breeding Place in the Poços de Caldas 36,0 % of Capillaria spp. and Eimeria spp. were identified. Considering all the birds evaluated for intestinal parasites in four zoos, Ramphastos spp. was the most prevalent bird for coccidium parasites although the number of birds of the Order Psittaciformes was much greater them the previous one. The length and width of oocysts from fecal samples of Ramphastos toco were measured (24,54 ?m ±1,17 x 23,83 ?m ±1,54) as well as the length and the width of two oocyst types found in Coragyps atratus (20,99 ?m ±1,24 x 17,59 ±1,78 ?m and 17,92 ?m ±0,74 x 15,59 ?m ±0,96). The statistical analysis of these measures showed significant difference between them. These measures and oocyst shapes compared with other measures and morphological oocyst structures described in the literature showed difference between them. Based on this information it was possible to conclude that they were considered new species of Eimeria not described previously. Three other species of Eimeria were also identified in the present study: E forresteri, E. Amazonae and E. Ochrocephalae. Some management proceedings in captive bird's environment for the prevention of coccidiosis and capilariosis were performed successfully after overall parasite identifications in the present study<br>Mestrado<br>Parasitologia<br>Mestre em Parasitologia

Made available in DSpace on 2015-04-22T22:06:28Z (GMT). No. of bitstreams: 1 sonia oliveira.pdf: 2531249 bytes, checksum: 7e7cb37ad572069870aa92e90ad2b74c (MD5) Previous issue date: 2013-08-30<br>FAPEAM - Fundação de Amparo à Pesquisa do Estado do Amazonas<br>Intestinal parasites are a serious public health problem in many regions of the world, especially in developing countries where rapid urban growth not keeping pace with demand for sanitation. The diagnostic tests used for detection of intestinal parasites have accuracy less than 100%, leading to diagnostic errors with false-negative results. Rarely discusses the optimal amount of readings for each stool sample to be examined by the method of sedimentation. Thus, this study aims to estimate the prevalence of intestinal parasites evaluating the strategy of increasing the sampling to detect parasites and establish possible correlations between parasites and environmental conditions. A group of 101 school students from the urban area of Manaus and 100 students of rural Rio Pardo, municipality of Presidente Figueiredo - Amazonas - Brazil were selected after sample calculation. Stool samples were collected and analyzed by the method of sedimentation using three stool samples and a total of nine readings blades. The prevalence of intestinal parasites was higher in urban areas with 90.10 %. The multiparasitism was 62.68 %. The common parasite that has been identified from both populations was Entamoeba coli rate of 34.49 % followed by 18.36 % Giardia intestinalis with statistical difference between two intestinal parasites proportion of urban and rural areas were : Giardia intestinalis (p - value = 0.0030 , OR 3.10 , CI 1.70 to 5.67) , Endolimax nana (p - value = 0.0000 , OR 37.44 , CI 13.92 to 100.71) , Ancylostoma spp. ( p - value = 0.0065 , OR 3.93 , CI 1.39 to 11.12) , Iodamoeba butschlii (p - value = 0.0007 , OR 7.01 , CI 2.00 to 24.64) , Hymenolepis nana (p - value = 0.0181 , OR 5.99 CI 1.29 to 27.76). There was no sex predominance in both populations. Only the rural statistical significance for variables 1-2 bedrooms (p - value = 0.0243), treatment of drinking water (p = 0.0227) and sewage disposal in the stream (p = 0.0447) . With the increased number of stool samples and reads blades there was also a considerable increase in positivity of most analyzes parasitological stool tests. The Wilcoxon test to determine the proportion of positive aspects of the readings of the blades had statistical significance in almost all intestinal parasites, except Endolimax nana (p - value = 0.059) and Hymenolepis nana (p - value = 0.317). From the fourth blade, the positive samples had an increased cumulative frequency ranging from 21% to five times coming. Procedures sedimentation increasing the number of samples and readings blades showed high ability to detect intestinal parasites.<br>As parasitoses intestinais constituem um grave problema de saúde pública em diversas regiões do mundo, principalmente nos países em desenvolvimento onde o rápido crescimento urbano não acompanha a demanda de saneamento básico. Os testes diagnósticos empregados na detecção de enteroparasitas possuem acurácia menor que 100%, levando a erros diagnósticos com resultados falso-negativos. Raramente se discute sobre a quantidade ideal de leituras para cada amostra de fezes a serem examinadas pelo método de sedimentação espontânea. Sendo assim, este estudo tem como objetivo estimar a prevalência de parasitoses intestinais avaliando a estratégia de aumentar a amostragem para detecção de parasitos e estabelecer possíveis correlações entre parasitoses e as condições socioambientais. Um grupo de 101 alunos da escola da zona urbana do município de Manaus e 100 alunos da zona rural de Rio Pardo, município de Presidente Figueiredo - Amazonas - Brasil foram selecionados após cálculo amostral. Foram coletadas amostras de fezes e analisadas pelo método de sedimentação espontânea utilizando três amostras de fezes e com um total de nove leituras de lâminas. A prevalência das parasitoses intestinais foi maior na área urbana com 90,10%. O multiparasitismo foi de 62,68%. O parasita comum que foi identificado a partir de ambas as populações foi a Entamoeba coli com taxa de 34,49% seguido da Giardia intestinalis com 18,36% Diferença estatística de entre duas proporção parasitos intestinais da área urbana e rural foram: Giardia intestinalis (p-valor = 0,0030; OR 3,10; IC 1,70-5,67), Endolimax nana (p-valor = 0,0000; OR 37,44; IC 13,92-100,71), Ancylostoma spp. (p-valor = 0,0065; OR 3,93; IC 1,39-11,12), Iodamoeba butschlii (p-valor = 0,0007; OR 7,01; IC 2,00-24,64), Hymenolepis nana (p-valor = 0,0181; OR 5,99 IC 1,29-27,76). Não foi observada predominância de sexo nas duas populações. Somente a área rural significância estatística para as variáveis de 1-2 quartos (p-valor = 0,0243), tratamento de água para consumo (p = 0,0227) e destino do esgoto no igarapé (p = 0,0447). Com o aumento do número de amostras de fezes e de leituras de lâminas houve também um aumento considerável na positividade da maioria das análises dos testes parasitológicos das fezes. O teste de Wilcoxon para verificar a proporção das positividades das leituras das lâminas teve significância estatística em quase todos parasitos intestinais, com exceção da Endolimax nana (p-valor = 0.059) e Hymenolepis nana (p-valor = 0,317). A partir da quarta lâmina, a positividade das amostras teve um aumento da frequência acumulativa que variou de 21% chegando a cinco vezes mais. Os procedimentos de sedimentação espontânea aumentando o número de amostras e de leituras de lâminas demonstraram alta capacidade para detecção de parasitas intestinais.

Orientador: Marlene Tiduko Ueta<br>Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia<br>Made available in DSpace on 2018-08-08T00:02:43Z (GMT). No. of bitstreams: 1 Alarcon_DiegoFernandes_M.pdf: 8141836 bytes, checksum: 830e38e5f19d73153a16ed4df89c73bc (MD5) Previous issue date: 2006<br>Resumo: Os helmintos intestinais de Lontra longicaudis (CARNIVORA, MUSTELIDAE) foram estudados, com os materiais fecais recolhidos de um pequeno riacho (Riacho Ribeirão Cachoeira) localizado em um fragmento florestal de Mata Atlântica, e de uma lagoa localizada no condomínio Notredame, ambos situados no município de Campinas, mais precisamente no distrito de Sousas. O objetivo foi relacionar, pelo levantamento de parasitas intestinais, aspectos tróficos e comportamentais de L. longicaudis à aquisição de parasitas. Em laboratório, o material fecal de cada local foi processado e analisado individualmente, utilizando-se os métodos qualitativos rotineiros de exame de fezes: sedimentação espontânea (HOFFMAN et al,. 1934) e Willis (WILLIS, 1921). Das 39 amostras coletadas em Ribeirão Cachoeira, 22 apresentaram resultados positivos para ovos e/ou larvas de helmintos (56,4%). Na lagoa do Notredame foram coletadas 29 amostras fecais, das quais 13 apresentaram resultados positivos para ovos ou larvas de helmintos (44,8%). Foram observados sete tipos morfológicos de ovos de helmintos e dois tipos de larvas de nematódeos. Dentre os ovos, os mais prevalentes, tanto no riacho (54,5% trematódeo; 22,7% Strongyloides) quanto na lagoa (46,2% trematódeo; 46,2% Strongyloides), foram o de um trematódeo e ovo de Strongyloides sp. Um dos tipos de larva encontrada foi o de Strongyloides sp, que foi a mais prevalente (54,5% riacho e 15,4% lagoa). Trata-se do primeiro registro de Strongyloides sp em lontras neotropicais<br>Abstract: The intestinal helminthes of the otter Lontra longicaudis (CARNIVORA, MUSTELIDAE) were studied. The fecal samples were collected in a creek (Ribeirão Cachoeira) situated in a forest fragment of Atlantic forest, and in a pond located in the Notredame house complex, both in the municipality of Campinas, Sousas district, São Paulo state, Brazil. The aim of the present study was to associate, through the intestinal parasites survey, trophic and behavioral aspects of L. longicaudis with the acquisition of parasites. The Faecal samples from each place was processed and analyzed individually, using the qualitative routine faecal methods: spontaneous sedimentation (HOFFMAN et al,. 1934) and Willis (WILLIS, 1921). From total 39 samples collected in Ribeirão Cachoeira creek, 22 were positive for eggs and /or larvae of helminthes (56,4%). In the Notredame house complex pond, samples from 29 total faecal samples collected, 13 showed positive results for eggs and /or larvae of helminthes (44,8%). Seven morphological types of helminthes eggs and two types of Nematode larvae were observed. Among the eggs, the most prevalent in the creek (54,5% trematode; 22,7% Strongyloides) and in the pond (46,2% trematode; 46,2% Strongyloides) were trematode egg and Strongyloides sp. egg. One of larvae found was of Strongyloides sp, type which was the most prevalent one (54,5% creek and 15,4% pond). This is the first occurrence of Strongyloides sp in neotropical otters<br>Mestrado<br>Mestre em Parasitologia

Les parasites du genre Cryptosporidium comprennent des espèces infectant le tractus gastro-intestinal ou respiratoire d’un grand nombre de vertébrés y compris l'homme. Ces protistes intracellulaires sont les agents d’une zoonose cosmopolite à transmission oro-fécale, la cryptosporidiose. Au vu des travaux de notre laboratoire, nous savons à présent que Cryptosporidium parvum est également capable d’induire des néoplasies digestives chez un modèle murin SCID (Severe Combined Immunodeficiency mice), traitées ou pas par la dexaméthasone. Alors que C. muris, une autre espèce de Cryptosporidium, induit une infection chronique non associée à des transformations néoplasiques.Pour toutes ces raisons, il nous est apparu intéressant d’effectuer un travail de thèse articulé autour de trois axes principaux, l’épidémiologie, la transmission et la pathogénicité du parasite Cryptosporidium. Nous nous sommes intéressés dans un premier temps à l’épidémiologie moléculaire et la biodiversité génétique de Cryptosporidium dans des populations humaines de la région du Nord-Liban. Ceci nous a permis de mettre en évidence une prévalence de 5% de Cryptosporidium chez la population générale avec une prédominance de C. hominis. Ce qui constituait les premières données épidémiologiques de la cryptosporidiose au Liban. Ensuite d’autres études nous ont permis de montrer que cette prévalence pouvait atteindre même 10% chez les patients symptomatiques et les enfants.Dans un second temps, nous avons voulu étudier le mode de transmission du parasite et les facteurs de risque pouvant y être associés. Pour ce faire, une recherche du parasite a été réalisée aussi bien au Liban qu’en France chez des animaux d’élevage, sauvages, de compagnie et en captivité. Une première étude a été réalisée chez des patients et des bovins du Nord-Liban. L’ensemble des données rapportées nous permettent de suggérer un mode de transmission de la cryptosporidiose majoritairement anthroponotique au Liban, mais les résultats du génotypage ne permettent pas d’exclure la présence d’une transmission zoonotique. D’autres études réalisées en France, notamment sur des échantillons de selles collectées auprès des zoos de la Palmyre (à Royan) et de Lille ont montré un taux de prévalence de Cryptosporidium spp inférieur à 1%. Ces animaux ne semblent donc pas être un réservoir potentiel de cette infection. Alors que chez les poissons sauvages, nous avons pu identifier la présence entre autre de l’espèce zoonotique C. parvum dans l’estomac et l’intestin des poissons. Ceci nous permet de considérer les poissons comme étant une source de contamination potentiel pour l’homme, l’animal mais également pour l’environnement.Enfin le troisième axe avait pour but d’étudier la pathogénicité de ce parasite. Pour commencer nous avons voulu explorer les mécanismes de la cancérogénèse induite par la souche IOWA de C. parvum au niveau de la région iléocæcale des souris SCID traitées par la dexaméthasone (SCID-D). Pour ce faire nous nous sommes intéressés à quatre marqueurs de voies de signalisation cellulaires impliquées dans la survenue de cancers colorectaux (APC, Bêta-caténine, P53 et K&#8208;ras). Nous avons ainsi pu montrer que la voie Wnt était impliquée dans ce processus. Ensuite nous avons voulu étudier l’association entre la pathologie cancéreuse et le parasitisme par Cryptosporidium chez l’homme. Une recherche du parasite a donc été réalisée dans des biopsies d’origines coliques et gastriques inclues en paraffine appartenant à des patients atteints ou non de cancers digestifs. Une différence significative a été rapportée entre la prévalence de la cryptosporidiose retrouvée chez la population de patients présentant des lésions cancéreuses (17%) et celle du groupe control constitué de patients non cancéreux mais présentant des symptômes (7%) p-value = 0.03. L’ensemble de ces données obtenues chez l’animal et chez l’homme montre que ce parasite a un impact important en santé humaine et animale<br>Parasites of the genus Cryptosporidium comprise species infecting the gastrointestinal or respiratory tract of a wide variety of vertebrates including humans. These intracellular protists are the agents of a cosmopolitan zoonosis, with féco-oral transmission, cryptosporidiosis. Recent work from our laboratory, showed that the zoonotic species Cryptosporidium parvum is capable to induce digestive neoplasia in a SCID Severe Combined Immunodeficiency Mice (SCID) model, treated or not with dexamethasone. However C. muris, another species of Cryptosporidium, induces chronic infection in this rodent model but is not associated with neoplastic transformation.For all these reasons, it seemed interesting to carry out a thesis project articulated around three different axes: epidemiology, transmission and pathogenesis of the Cryptosporidium infection. We focused initially on the molecular epidemiology and genetic biodiversity of this parasite among human populations in North Lebanon. We found a Cryptosporidium prevalence of 5% among the general population, being C. hominis the predominant species. This prevalence could reach until 10% in symptomatic patients and children. This is the first epidemiological data about cryptosporidiosis in this country.Secondly, we studied the transmission routes and the main risk factors associated with the transmission of this parasite. To do this, a first study was conducted in parallel among animal populations in North Lebanon and France. The reported data suggest a predominance of an anthroponotic route of transmission for cryptosporidiosis in Lebanon, but the results of genotyping does not exclude the presence of zoonotic transmission. Other studies conducted in France, especially based on collection of stool samples in the zoos of Palmyre (Royan) and Lille showed that Cryptosporidium spp were present in less than 1% of captivity animals. The low prevalence strongly demonstrates that these animals play a negligible role as potential reservoirs of infection. While in wild fish, we could identify the presence of C. parvum, a zoonotic species, in the stomach and the gut of fish. These data suggest that the fish could be a natural host of C. parvum and a potential source of contamination for humans, animals but also for the environment.Finally, the third topic aimed to study the pathogenicity of this parasite. Firstly, we explored metabolic pathways potentially involved in the development of C. parvum-induced ileo-caecal oncogenesis in the SCID model treated with dexamethasone (SCID-D). We searched for alterations in genes or proteins commonly involved in cell cycle, differentiation or cell migration, such as &#946;-catenin, Apc, E-cadherin, Kras and p53. We were able to show that the Wnt pathway was involved in this process. Finally, we wanted to study the association between digestive cancer and parasitism with Cryptosporidium in humans. Cryptosporidium molecular detection was therefore carried out in colonic and gastric biopsies belonging to patients with and without digestive cancers of recent diagnosis collected in North Lebanon. A statistically significant difference was observed between the prevalence of cryptosporidiosis found among the population of patients with digestive cancer (17%) and that of the control group consisting of non-cancer patients but with digestive symptoms (7%) (p-value = 0.03). All these data obtained in animals and humans strengthens the importance of this parasite in public health

Made available in DSpace on 2014-11-10T11:09:55Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-02-21Bitstream added on 2014-11-10T11:57:43Z : No. of bitstreams: 1 000785387.pdf: 1004100 bytes, checksum: 60651dd50c503ca75c5c206fb2da89b0 (MD5)<br>A resistência anti-helmíntica em bovinos se tornou um problema global, e no Brasil tem sido relatada em alguns estados. Devido à necessidade de métodos mais precisos de diagnóstico da resistência anti-helmíntico, o objetivo deste trabalho foi avaliar duas técnicas (McMaster e Flotac) e duas fórmulas matemáticas (FECRT1 - de acordo com a média de OPG e FECRT2 - de acordo com o número total de ovos contados) para calcular o teste de redução de ovos nas fezes (FECRT). O anti-helmíntico testados foram ivermectina (0,2 mg/kg), moxidectina (0,2 mg/kg) , sulfóxido de albendazol (2,5 mg/kg) e fosfato de levamisol (4,7 mg/kg) , administrada na dose recomendada pelo fabricante. Dez propriedades foram avaliadas, quatro visitas em cada, pré-tratamento (D - 2), processamento (D 0), 10 dias pós-tratamento (D10) e 28 dias póstratamento (D28). Em todas as propriedades foi observada resistência a um ou mais agentes anti-helmínticos testados, independentemente da técnica ou da fórmula matemática avaliada. Os gêneros/espécies encontradas com resistência a um ou mais anti-helmínticos foram Haemonchus placei, Cooperia spp., Trichostrongylus spp. e Oesophagostomum radiatum. Em conclusão, a resistência anti-helmíntica está presente em todos os rebanhos avaliados. A técnica Flotac e a fórmula matemática FECR2 foram mais precisas do que a técnica de McMaster e fórmula FECR1. Há necessidade de maior conscientização dos produtores sobre o uso de anti-helmínticos. Portanto, em estudos futuros de resistência anti-helmíntica em bovinos, recomenda-se a utilização do método de diagnóstico com uma maior precisão, como é o método Flotac. Na interpretação dos resultados, nosso estudo indica que o cálculo pela fórmula FECR2, o que elimina o uso de um grupo de controle, deve ser recomendada<br>The anthelmintic resistance in cattle has become a global problem, and in Brazil it has been reported in some states. Due to the need for more accurate methods of diagnosis of anthelmintic resistance the purpose this work was to evaluate two techniques (McMaster and Flotac) and two mathematical formulas (FECRT1 - according to the average EPG and FECRT2 - according to the total number of eggs counted) to calculate the faecal egg count reduction test (FECRT) in the diagnosis of anthelmintic resistance in cattle. The anthelmintic tested were ivermectin (0.2 mg/kg), moxidectin (0.2 mg/kg), albendazole sulfoxide (2.5 mg/kg) and levamisole phosphate (4.7 mg/kg), administered at the dose recommended by the manufacturer. Ten farms were assessed, four visits in each pre-treatment (D -2), processing (D 0), 10 days post-treatment (D 10) and 28 days post-treatment (D 28). In all of the properties was observed resistance to one or more anthelmintics tested, regardless of the technique (McMaster or Flotac) or FECRT the mathematical formula (1 and 2). The genera/species found with resistance to one or more anthelmintics were Haemonchus placei, Cooperia spp., Trichostrongylus spp. and Oesophagostomum radiatum. In conclusion, the anthelmintic resistance is present in all herds evaluated. The Flotac technical and mathematical formula FECR2 were more accurate than the McMaster technique and formula FECR1. Need is greater awareness of the producers on the use of anthelmintics. Therefore, in future studies on anthelmintic resistance in cattle, use of the method of diagnosis with higher accuracy, it is recommended, as is the case FLOTAC method. In interpreting the results, our study indicates that the calculation by formula FECR2 %, which eliminates the use of a control group, should be recommended

Orientador: Alessandro Francisco Talamini do Amarante<br>Banca: Cyntia Ludovico Martins<br>Banca: Helder Louvandini<br>Resumo: A resistência anti-helmíntica em bovinos se tornou um problema global, e no Brasil tem sido relatada em alguns estados. Devido à necessidade de métodos mais precisos de diagnóstico da resistência anti-helmíntico, o objetivo deste trabalho foi avaliar duas técnicas (McMaster e Flotac) e duas fórmulas matemáticas (FECRT1 - de acordo com a média de OPG e FECRT2 - de acordo com o número total de ovos contados) para calcular o teste de redução de ovos nas fezes (FECRT). O anti-helmíntico testados foram ivermectina (0,2 mg/kg), moxidectina (0,2 mg/kg) , sulfóxido de albendazol (2,5 mg/kg) e fosfato de levamisol (4,7 mg/kg) , administrada na dose recomendada pelo fabricante. Dez propriedades foram avaliadas, quatro visitas em cada, pré-tratamento (D - 2), processamento (D 0), 10 dias pós-tratamento (D10) e 28 dias póstratamento (D28). Em todas as propriedades foi observada resistência a um ou mais agentes anti-helmínticos testados, independentemente da técnica ou da fórmula matemática avaliada. Os gêneros/espécies encontradas com resistência a um ou mais anti-helmínticos foram Haemonchus placei, Cooperia spp., Trichostrongylus spp. e Oesophagostomum radiatum. Em conclusão, a resistência anti-helmíntica está presente em todos os rebanhos avaliados. A técnica Flotac e a fórmula matemática FECR2 foram mais precisas do que a técnica de McMaster e fórmula FECR1. Há necessidade de maior conscientização dos produtores sobre o uso de anti-helmínticos. Portanto, em estudos futuros de resistência anti-helmíntica em bovinos, recomenda-se a utilização do método de diagnóstico com uma maior precisão, como é o método Flotac. Na interpretação dos resultados, nosso estudo indica que o cálculo pela fórmula FECR2, o que elimina o uso de um grupo de controle, deve ser recomendada<br>Abstract: The anthelmintic resistance in cattle has become a global problem, and in Brazil it has been reported in some states. Due to the need for more accurate methods of diagnosis of anthelmintic resistance the purpose this work was to evaluate two techniques (McMaster and Flotac) and two mathematical formulas (FECRT1 - according to the average EPG and FECRT2 - according to the total number of eggs counted) to calculate the faecal egg count reduction test (FECRT) in the diagnosis of anthelmintic resistance in cattle. The anthelmintic tested were ivermectin (0.2 mg/kg), moxidectin (0.2 mg/kg), albendazole sulfoxide (2.5 mg/kg) and levamisole phosphate (4.7 mg/kg), administered at the dose recommended by the manufacturer. Ten farms were assessed, four visits in each pre-treatment (D -2), processing (D 0), 10 days post-treatment (D 10) and 28 days post-treatment (D 28). In all of the properties was observed resistance to one or more anthelmintics tested, regardless of the technique (McMaster or Flotac) or FECRT the mathematical formula (1 and 2). The genera/species found with resistance to one or more anthelmintics were Haemonchus placei, Cooperia spp., Trichostrongylus spp. and Oesophagostomum radiatum. In conclusion, the anthelmintic resistance is present in all herds evaluated. The Flotac technical and mathematical formula FECR2 were more accurate than the McMaster technique and formula FECR1. Need is greater awareness of the producers on the use of anthelmintics. Therefore, in future studies on anthelmintic resistance in cattle, use of the method of diagnosis with higher accuracy, it is recommended, as is the case FLOTAC method. In interpreting the results, our study indicates that the calculation by formula FECR2 %, which eliminates the use of a control group, should be recommended<br>Mestre

Made available in DSpace on 2015-03-26T13:46:53Z (GMT). No. of bitstreams: 1 texto completo.pdf: 126733 bytes, checksum: af1fa3f739aff7ae6ddf9eed2f8af7f2 (MD5) Previous issue date: 2004-12-15<br>The objective of this work was to evaluate the individual efficacy of the fenbendazole and the pyrantel pamoate against intestinal nematodes of dogs. Thirty six nestlings of dogs were used, being eighteen males and eighteen females up to six months of age, coming of the kennel of the Department of Veterinary Medicine of the Federal University of Viçosa, selected through fecal exams of Willis, simple sedimentation and count of eggs per gram of feces (EPG). being used the animals that presented mixed infection for Ancylostoma sp. and Toxocara canis. The animais were distributed in three groups of twelve animals, being a control group, a group treated with fenbendazole, 100 mg for Kg of body weight, orally, in an unique dose, and the other with pyrantel pamoate, 5 mg for Kg of body weight, orally, in an unique dose. During the whole experimental period, the dogs received commercial ration and water ad libitum . Fecal exams of Willis, simple sedimentation and EPG realized in the day of the treatment (0), 24 hours after the treatments (1) and in the 3, 5 and 7 after the treatments. In the seventh day, the animals were killed and necropsied for collection of the adult worms and the mucosa of the gastrointestinal tract was submitted to digestion process, in hydrochloric acid to 3%, for research of immature stages. The results showed an efficacy of 99,89% of the pyrantel pamoate against the A. caninum with reduction of 100% of EPG in the seventh day after the treatment, and against the T. canis the effectiveness was of 71,65%, with reduction of 80,73% of OPG. The fenbendazole showed an efficacy of 93,19% against the A. caninum, with reduction of 96,22% in the EPG and an efficacy of 82,1% against the T. canis, reducing the EPG in 95,71%. The results demonstrate the efficacy for the control of the A. caninum with the two drugs, with higher effective of the pyrantel pamoate. To the T. canis control the drugs shower lower efficacy when administered separately in the doses recommended commercially.<br>Este trabalho teve como objetivo avaliar a eficácia individual do fembendazol e do pamoato de pirantel contra nematóides intestinais de cães. Para tal, foram utilizados 36 filhotes de cães, sendo 18 machos e 18 fêmeas com menos de seis meses de idade , provenientes do canil do Departamento de Veterinária da Universidade Federal de Viçosa, selecionados por meio de exames coprológicos de Willis, sedimentação simples e contagem de ovos por grama de fezes (OPG). Foram utilizados os animais que apresentaram infecção mista por Ancylostoma sp. e Toxocara canis. Estes foram distribuídos em três grupos de doze animais, sendo um grupo controle, um tratado com fembendazol, 100 mg por Kg de peso vivo, por via oral, em dose única, e o outro com pamoato de pirantel, 15mg por Kg de peso vivo, por via oral, em dose única. Durante todo o período experimental, os cães receberam ração comercial e água à vontade. Foram realizados exames coprológicos de Willis, sedimentação simples e contagem de ovos por grama de fezes (OPG), no dia do tratamento (dia 0), 24 horas após os tratamentos (dia 1) e nos dias 3, 5 e 7 pós-tratamento. No sétimo dia, os animais foram sacrificados e necropsiados para coleta dos vermes adultos e as mucosas do trato gastrintestinal passaram por um processo de digestão, em acido clorídrico a 3%, para pesquisa de estádios imaturos. Os resultados obtidos mostraram que houve uma eficácia de 99,89% do pamoato de pirantel contra o A. caninum e uma redução de 100% do OPG no sétimo dia após o tratamento, já contra o T. canis a eficácia foi de 71,63%, com redução de 80,73% do OPG. O fembendazol apresentou uma eficácia de 93,19% contra o A. caninum, com redução de 96,22% do OPG e uma eficácia de 82,1% contra o T. canis, reduzindo o OPG em 95,71%. Para o controle do A. caninum, as duas drogas se mostraram eficazes, com maior eficácia do pamoato de pirantel, mas, para o T. canis as drogas foram pouco eficazes quando administrados isoladamente nas doses comercialmente recomendadas.

Orientador: Semíramis Guimarães Ferraz Viana<br>Resumo: Giardia duodenalis é um dos principais agentes etiológicos de diarreia infecciosa, sobretudo em crianças em idade pré-escolar que vivem em comunidades de baixa renda. Estudos da diversidade genética de G. duodenalis ampliaram o conhecimento da epidemiologia nas infecções humanas, entretanto um dos temas mais interessantes e menos conhecidos é a possível interação de Giardia com o microbioma do hospedeiro e com patógenos concomitantes. No presente estudo, avaliou-se a composição e a diversidade da comunidade bacteriana de crianças saudáveis e crianças com diarreia, parasitadas por Giardia e outros protozoários intestinais. Os isolados de Giardia obtidos nessa população foram caracterizados geneticamente. Amostras de fezes foram obtidas de 181 crianças de 0 a 6 anos de idade, das quais 156 crianças hígidas atendidas em centros de educação infantil e 25 crianças com diarreia atendidas no PS Infantil Municipal. Cada amostra de fezes foi processada para o exame microscópico e submetida à extração de DNA a ser empregado em duas etapas distintas: (1) amplificação e sequenciamento Sanger para a caracterização genética de Giardia e o diagnóstico de Blastocystis sp, Dientamoeba fragilis, Enterocytozoon bieneusi e Cryptosporidium spp. e (2) amplificação do gene 16s RNA ribossomal e sequenciamento de nova geração (plataforma Illumina MiSeq) para a caracterização da microbiota intestinal. Giardia (36,5%) e Blastocystis (41,7%) foram os parasitas mais prevalentes. A caracterização genética... (Resumo completo, clicar acesso eletrônico abaixo)<br>Abstract: Giardia duodenalis is one of the major etiological agents of infectious diarrhea, especially in preschool children living in low-income settings. Studies focused on genetic diversity of G. duodenalis have provided insights for a better understanding of epidemiology in human infections. However, one of the most interesting and least known issues is the possible interplay between Giardia and the host microbiome and concomitant pathogens. In this work, we evaluated the diversity and composition of bacterial community of healthy children and children presenting with diarrhea infected by Giardia and/or other intestinal protozoa. In addition, Giardia isolates infecting this population were genotyping. A total of 181 stool samples from children aged 0 to 6 years old (156 from daycare children and 25 from diarrheic children attending in an emergence pediatric center) were tested by microscopic examination and submitted to DNA extraction for the following steps: (1) conventional PCR/sequencing for Giardia genotyping and the diagnosis of Blastocystis sp, Dientamoeba fragilis, Enterocytozoon bieneusi and Cryptosporidium spp and (2) next-generation sequencing (Illumina MiSeq) based analysis of intestinal microbiota. Giardia (36.5%) and Blastocystis (41.7%) were the most prevalent parasites. Analysis of Giardia sequences retrieved from 61 isolates revealed infections by assemblages A (31%), B (69%) and mixed infections A+B (3%). Metagenomic analyzes revealed similarity of bacterial microb... (Complete abstract click electronic access below)<br>Doutor

Submitted by Viviane Lima da Cunha (viviane@biblioteca.ufpb.br) on 2017-07-07T14:24:58Z No. of bitstreams: 1 arquivototal.pdf: 2296320 bytes, checksum: 5609e539752b8a82d04f4ef213001d17 (MD5)<br>Made available in DSpace on 2017-07-07T14:24:58Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 2296320 bytes, checksum: 5609e539752b8a82d04f4ef213001d17 (MD5) Previous issue date: 2015-12-17<br>Intestinal parasites diseases are responsible for serious problems in public health, intensely related to health conditions. They generate economic and social harm to the population, reducing their quality of life in developing countries and are also responsible for high mortality rates especially where health is unassisted. This study aided to analyze the association between these infections and clinical variables, haematological and epidemiological and introduce Decision Making considered by Logistic Regression model. This is an observational, analytical, cross-sectional study on the prevalence of intestinal parasites in patients treated at the Gastroenterology clinic of University Hospital Lauro Wanderley, João Pessoa, Paraíba, under the exclusive public service of Brazil’s Unified Public Health System (SUS). The data were collected by the analysis of the records from October 2014 to May 2015. Among the 370 records analyzed, through inclusion and exclusion criteria, totaled 34. The data were organized and tabulated in a spreadsheet and later submitted, through the statistical software R version 2.15.1, to a descriptive analysis and later to an associative analysis between variables as well as the Logistic Regression. The results showed that females were more affected by parasites and the average group-age of subjects was 56 years old and in general from João Pessoa. The helminth was most prevalent when compared to protozoa, being predominantly Schistosoma mansoni. The most widely used methods for fecal examinations were Hoffman and Kato-katz. Among the results of biochemical analysis, most individuals presented levels of leukocytes out of the reference values. Through the relative risk, protective factors were observed between gastrointestinal symptoms, diabetes, hypertension, and diuretic in relation to the presence of the parasite and risk factors between the independent variables and the outcome variable have been gastrointestinal comorbidities, antiparasitic medicine and gastroprotetor.<br>As parasitoses intestinais são doenças responsáveis por graves problemas na saúde pública, relacionadas às condições sanitárias. Geram danos econômicos e sociais para a população, diminuindo a qualidade de vida em países que estão em desenvolvimento, sendo também responsável por elevados índices de mortalidade, principalmente onde a saúde é desassistida. O objetivo deste estudo é analisar a associação entre estas infecções e variáveis clínicas, hematológicas. Trata-se de um estudo observacional, analítico, sobre a prevalência de enteroparasitoses em pacientes atendidos no ambulatório de Gastroenterologia do Hospital Universitário Lauro Wanderley, município de João Pessoa, Estado da Paraíba, com atendimento público exclusivo no âmbito do Sistema Único de Saúde (SUS). Os dados foram coletados através da análise de prontuários no período de outubro de 2014 a maio de 2015. Os dados foram organizados e tabulados numa planilha e posteriormente submetidos ao software estatístico R versão 2.15.1 a fim de se realizar uma análise descritiva e posteriormente a uma análise associativa entre as variáveis bem como a Regressão Logística. Os resultados mostraram que os indivíduos do sexo feminino foi mais afetado por parasitoses, a faixa etária média dos indivíduos foi de 56 anos e em geral provenientes de João Pessoa. Os helmintos foram mais prevalentes em relação aos protozoários, sendo predominantemente Schistosoma mansoni. Os métodos mais utilizados para exames coproparasitológicos foram o Hoffman e Kato-katz. Dentre os resultados das análises bioquímicas, grande parte dos indivíduos apresentou níveis de leucócitos fora dos valores de referência. Através do risco relativo foram observados fatores de proteção entre os sintomas gastrointestinais, diabetes, HAS, e diurético em relação à presença do parasita e fatores de risco entre as variáveis independentes; e a variável desfecho têm-se comorbidades gastrointestinais, medicamento antiparasitário e gastroprotetor.

Made available in DSpace on 2016-12-23T13:56:08Z (GMT). No. of bitstreams: 1 Dissertacao de Steveen Rios Ribeiro.pdf: 4201771 bytes, checksum: 56acf4462c81abd472caf4c84eb8dd80 (MD5) Previous issue date: 2011-03-18<br>Coordenação de Aperfeiçoamento de Pessoal de Nível Superior<br>Introduction. The diagnosis of intestinal parasites can be accomplished by different techniques of parasitological stool examinations (EPF), each with advantages and disadvantages according to the parasite to be identified. Aims. The aim of the present study was to compare traditional techniques of EPF (Spontaneous Sedimentation in Tube, Kato-Katz and Baermann-Moraes) with more sophisticated techniques (stool culture, detection of coproantigen and PCR) and two commercially available kits in Brazil (Paratest® and TF-Test ®). Material and Methods. Faecal samples from 160 individuals, a total of 356 samples, were analyzed by traditional techniques of EPF (Spontaneous Sedimentation in Tube, Kato-Katz and Baermann-Moraes), culture of fresh faeces and sediment of feces, detection of coproantigen, PCR for detection and differentiation of the complex Entamoeba histolytica/Entamoeba dispar and two commercially available kits in Brazil (Paratest® and TF-Test®). Results. The technique of Spontaneous Sedimentation in Tube had better sensitivity (80.43% of occurrences) for the diagnosis of eggs and larvae of helminths in feces. The Kato-Katz was better for identification of eggs of S. mansoni (100% of cases). The method of Baermann-Moraes found larvae of Strongyloides stercoralis detected similarly to spontaneous sedimentation. For the diagnosis of G. lamblia, C. parvum and E. histolytica, kits for detection of coproantigen showed better sensitivity. The stool culture showed excellent results for identification of Blastocystis hominis. The PCR was able to identify and differentiate the amoebas of the complex E. histolytica/ E. dispar. Both commercial kits showed lower sensitivity than the spontaneous sedimentation for the diagnosis of helminths, but Paratest® was the technique that has shown better sensitivity for protozoa (66.66% of cases). Conclusion. For the diagnosis of helminths, the Spontaneous Sedimentation Method in Tube showed better results, except for detection of eggs of S. mansoni, best detected by Kato-Katz. For the diagnosis of G. lamblia, E. histolytica and Cryptosporidium parvum, kits for detection of coproantigen were better. However, the kit for the diagnosis of E. histolytica does not detect the species E. dispar, best identified by the ABSTRACT PCR. The commercial kits (Paratest® and TF-Test ®), while facilitating the collection and preservation of the sample, have lower sensitivity than the Spontaneous Sedimentation method in Tube<br>O diagnóstico de parasitoses intestinais pode ser realizado por diferentes técnicas de Exame Parasitológico de Fezes (EPF), cada uma com vantagens e desvantagens de acordo com o parasito a ser identificado. Objetivos. Comparar técnicas tradicionais de EPF (Sedimentação Espontânea em Tubo, Kato-Katz e Baermann-Moraes) com técnicas mais sofisticadas (cultura de fezes, detecção de coproantígenos e PCR) e dois kits comerciais disponíveis no Brasil (Paratest® e TF-Test®). Material e Métodos. Amostras de fezes de 160 indivíduos, totalizando 356 amostras, foram analisadas por técnicas tradicionais de EPF (Sedimentação Espontânea em Tubo, Kato-Katz e Baermann-Moraes), cultura de fezes frescas e de sedimento de fezes, técnicas para detecção de coproantígenos nas fezes, PCR para detecção e diferenciação de Entamoeba do complexo histolytica/dispar e dois kits comerciais disponíveis no Brasil (Paratest® e TF-Test®). Resultados. A técnica de Sedimentação Espontânea em Tubo demonstrou melhor sensibilidade (80,43% das o corrências) para o diagnóstico de ovos e larvas de helmintos nas fezes. O Kato-Katz foi melhor para identificação de ovos do S. mansoni (100% das ocorrências). Já o método de Baermann-Moraes detectou larvas de Strongyloides stercoralis de modo semelhante à sedimentação espontânea. Para o diagnóstico de G. lamblia, C. parvum e E. histolytica, os kits de coproantígenos demonstraram melhor sensibilidade. A cultura de fezes mostrou excelentes resultados para identificação de Blastocystis hominis. O PCR conseguiu identificar e diferenciar as amebas do complexo E. histolytica/E. dispar. Os dois kits comerciais apresentaram sensibilidade mais baixa do que a Sedimentação Espontânea em Tubo para o diagnóstico de helmintos, mas o Paratest® foi a técnica que demonstrou melhor sensibilidade para protozoários (66,66% das ocorrências). Conclusão. Para o diagnóstico de helmintos o método de Sedimentação Espontânea em Tubo foi o que mostrou melhor resultado, exceto para detecção de ovos de S. mansoni, melhor detectados pelo Kato-Katz. Os kits de detecção de coproantígenos foram melhores para diagnóstico de G. lamblia, Cryptosporidium parvum e E. histolytica. Entretanto, o kit para diagnóstico de E. histolytica não detecta a espécie E. dispar, melhor identificada pelo PCR. Os kits comerciais (Paratest® RESUMO e TF-Test®), apesar de facilitarem a coleta e conservação da amostra, têm menor sensibilidade do que a Sedimentação Espontânea em Tubo

Escherichia coli enteroinvasora (EIEC) e Shigella sp causam disenteria bacilar que é caracterizada pela invasão e destruição da mucosa do cólon humano. Amostras de EIEC possuem características bioquímicas, genéticas patogênicas semelhantes às espécies de Shigella, porém a doença causada por EIEC se apresenta numa forma mais branda e autolimitante. As células do epitélio intestinal participam ativamente da imunidade da mucosa, expressando e secretando uma série de mediad ores inflamatórios como citocinas, quimiocinas, moléculas de adesão e óxido nítrico. Para melhor entendimento da patogênese de EIEC, estudamos a resposta inflamatória modulada por este microrganismo em células epiteliais intestinais da linhagem Caco-2, comparando-a com Shigella flexneri. Células Caco-2 foram infectadas com EIEC ou S. flexneri por diferentes intervalos de tempo, para posterior analise da capacidade de invasão e disseminação bacterianas (UFC, PLAQUE ASSA Y), indução de morte celular (FACS), analise relativa de genes envolvidos no reconhecimento bacteriano e na resposta inflamatória (RT-PCR, RPA), dosagem de citocinas e quimiocinas pró-inflamatórias (ELISA) e óxido nítrico (GRIESS). Neste trabalho foi possível observar que: (i) a capacidade de disseminação e (ii) a indução da morte celular em células Caco-2 foi significativamente maior na infecção por S. flexneri do que EIEC; (iii) há diferenças em relação à expressão relativa de genes das células Caco-2 envolvidos no reconhecimento das duas cepas bacterianas. Foi evidenciado o papel essencial dos receptores intracelulares no reconhecimento bacteriano das células Caco-2, sendo a expressão relativa do mRNA do receptor intracelular Nod1 foi maior para EIEC quando comparado com S. flexneri; (iv) há diferenças significativas na cinética de produção de NO pelas células Caco¬2 infectadas, em que EIEC induziu mais precocemente a produção de NO quando comparado com S. flexneri. Estes dados sinalizam que as células epiteliais intestinais reconhecem e respondem de forma diferente frente a essas duas espécies bacterianas, apresentando uma resposta inflamatória mais eficiente no controle da infecção induzida por EIEC.<br>Escherichia coli enteroinvasive (EIEC) and Shigella sp cause bacillary dysentery which is characterized by the invasion and destruction of the human colon mucosa. Samples of EIEC have characteristics biochemical, genetic and pathogenic similar to those of Shigella species, however the disease caused by EIEC is more lenient. The cells of the intestinal epithelium actively participate in the mucosal immunity by expression and production of several inflammatory mediators such as cytokines, chemokines, adhesion molecules and nitric oxide. For better understanding of the EIEC pathogenesis, we studied the inflammatory response modulated by this microorganism in intestinal epithelial cells Caco-2, comparing it with Shigella flexneri. Caco-2 cells were infected with EIEC or S. flexneri during different intervals of time and analyzed the invasiveness and spread bacteria capacity (CFU, PLAQUE ASSAY), induction of cell death (FACS), analysis of genes involved in the recognition of bacterial and inflammatory response (RT-PCR, RPA), production of pro-inflammatory cytokines and chemokines (ELISA) and nitric oxide (NO) (GRIESS). In this work was possible to observe that: (i) the ability to spread and (ii) the induction of cell death in Caco-2 cells was significantly higher in S. flexneri infection than EIEC, (iii) there are differences regarding the relative expression of genes of Caco-2 cells involved in the recognition of two bacterial strains. It was highlighted the essential role of intracellular receptors in recognition of bacterial by Caco-2 cells, and the expression of mRNA of the intracellular receptor Nod 1 was higher for EIEC when compared with S. flexneri, (iv) there are significant differences in the kinetics of NO production by Caco-2 infected cells, EIEC induced a early NO production when compared with S. flexneri. These data indicate that the intestinal epithelial cells recognize and respond in a different way to these bacterial species and induce an inflammatory response more efficient in control of the infection induced by EIEC.

Early innate response of intestinal epithelial cells is the first line defense against enteric pathogens. Toxoplasma gondii infections acquired naturally via the peroral route, encounter intestinal epithelial cells early post-infection. Although the population structure of T. gondii is known to be highly clonal, clinical strains of T. gondii have been classified into three genotypes based on their virulence. In this study we investigated whether human intestinal epithelial cell immune response to T. gondii is virulence dependent. We demonstrated distinct virulence of the three T. gondii genotype strains evaluated in human intestinal epithelial cells by their capacity to replicate and induce host cell cytotoxicity. The early host innate mechanisms such as activation of signaling pathways and induction of innate effectors were likewise differentially elicited by the three T. gondii strains. Low levels of TLR dependent NF-kB activation and a failure to rapidly up-regulate innate cytokine and chemokine genes was observed after virulent Type I strain infection. In contrast, early innate response to the less virulent Type II strain was rapid, efficient and led to high levels of IL-8 and IL-6 secretion, whereas response to Type III parasites was intermediate. Early expression of b-defensin 2 gene was suppressed specifically by virulent Type I strain and its activation prior to infection in intestinal epithelial cells led to decreased parasite viability. These findings provide evidence for T. gondii strain virulence dependent down-modulation of early human intestinal epithelial cell innate responses and highlight the importance of these cells in host defense against this infection.<br>Doctorat en Sciences biomédicales et pharmaceutiques<br>info:eu-repo/semantics/nonPublished

Les systèmes d'élevage des petits ruminants au pâturage sont les plus répandus en zone tropicale humide mais l'exposition aux strongles gastro-intestinaux entraîne des pertes importantes de production. L'objectif de ce travail est d'évaluer un système de gestion mixte du pâturage associant diverses espèces animales permettant à priori d'accroître les performances via la réduction du parasitisme et!ou une meilleure alimentation. L'étude de la littérature a conduit à une méta-analyse confirmant de meilleures performances individuelles ou à l'hectare pour les ovins en mixte. Préalablement un dispositif visant à vérifier la non-transmission des larves d'Haemonchus contortus des caprinsaux génisses Créoles a été validé. Un second dispositif avec des chevrettes conduites en mixte, infestées ou pas par Haemonchus comortus (Ml et MnI) ont été comparées à des chevrettes contrôles infestées ou non (CI et CnI). Ce dispositif a été conduit pendant 2 ans avec des mesures du couvert pâturé, des mesures individuelles des quantités ingérées, de la digestibilité, du parasitisme et de la croissance. De plus fortes croissances ont étéobservées en pâturage mixte que les chevrettes soient infestées ou pas (43.25 et 31.68 glj pour MDI et MI vs. 32.44 et 17.91 glj) avec une moindre biomasse. Les mesures d'ingestion et d'ingéré digéré ont été corrélées aux croissances des chevrettes contrairement aux variables parasitaires et met en évidence le rôle de l'alimentation et de la résilience dans le bénéfice du pâturage mixte. Une autre gestion intégrée du pâturage a été étudiée basée sur l'utilisation du vermicompost et s' ant sur les mêmes leviers d'action: l'alimentation et le itisme<br>Systems of small ruminant grazing are most prevalent in the humid tropics but this environment exposes animals to gastro-intestinal with production losses. The objective ofthis thesis is to evaluate a system ofmixed management ofthe pasture combining various animal species, which allows a priori to increase animal performance by reducing the parasitism and/or better nutrition. The study of literature bas led to a meta-analysis confirming the better individual performances or calculated per ha for sheep reared mixed. The first experiment validated the non-transmission of Iarvae ofHaemonchus contortus from goats to heifers. In a second experiment, goats mixed with heifers, infested or not with Haemonchus contortus (Ml vs MnI) were compared with controls goats, reared alone, infested or not (CI and CNI). This deviee with a continuous driving was studied for two years with measurements on the sward, individual measures of intake, digestibility, parasitism and growth ofthe goats. The highest growth rates were measured with the mixed pasture as goats are infested or not (43.25 and 31.68 g / d for MN! and MI vs. 32.44 and 17,91 g / d) with lower biomass. The herbage intake and the digested herbage intake were well correlated to the growth rate, unIike parasitic variables, and highlight the role of diet in the benefit of mixed driving for goats. Therefore, the infested goats with Haemonchus, expressed greater resilience with improved growth performance compared with controls. Parallel to the study ofmixed pasture, another integrated management of grazing was studied, based on the use of vermicompost and relying on the same levers. namelv throuzh feedinz and parasitism

Les microsporidies intestinales sont reconnues comme etant les principaux agents responsables de diarrhee chronique severe chez les sujets infectes par le virus de l'immunodeficience humaine (vih), leur taux de prevalence atteignant 39%. Les deux especes responsables sont enterocytozoon bieneusi et encephalitozoon intestinalis. La tres petite taille des spores de ces parasites explique les difficultes rencontrees lors de leur detection. La mise au point de techniques de diagnostic sensibles et specifiques de ces nouveaux agents pathogenes a suscite un effort particulier sur le plan international. Notre travail s'est inscrit dans cet objectif. Nous avons entrepris deux demarches : 1. La recherche d'anticorps specifiques des microsporidies intestinales dans le serum des patiens vih+ ayant une microsporidiose intestinale. Nous avons mis en evidence la presence d'anticorps diriges contre enc. Intestinalis chez des patients vih+ infectes par enc. Intestinalis pourtant severement immunodeprimes. Les antigenes du filament polaire, identifies par immunocytochimie, presentent un interet particulier car ils sont reconnus par le serum de ces patients comme par celui de donneurs de sang. Ces resultats suggerent l'existence d'encephalitozoonoses dans la population immunocompetente. 2. La mise au point d'une technique de detection des spores dans les selles de ces patients. Nous proposons deux nouvelles methodes de diagnostic qui, par leur specificite et leur sensibilite, sont complementaires des techniques deja existante. Il s'agit d'une part, d'une technique de double marquage, combinant une reaction d'immunofluorescence indirecte classique avec un serum polyclonal anti-enc. Intestinalis et le marquage a l'uvitex 2b, et d'autre part, d'une technique de pcr directement applicable dans les selles des patients. Cette deuxieme technique, aussi sensible que les techniques classiques, permet en plus la differenciation des deux especes de microsporidies.

Blastocystis est un protozoaire anaérobie trouvé dans le tube digestif de l’homme et de nombreux animaux. Il est à ce jour le parasite intestinal le plus fréquemment retrouvé dans les selles humaines. Dix-sept sous-types (ST1 à ST17) ont été décrits en se basant sur la comparaison des séquences du gène de l’ARNr 18S. L’infection à Blastocystis est associée à une variété de troubles gastro-intestinaux et plusieurs études suggèrent une corrélation entre la pathogénicité et le ST du parasite. Trois différents axes de recherche ont été développés. Le premier s’est focalisé sur la prévalence et la biodiversité génétique de ce parasite dans les populations humaines. Des études épidémiologiques ont été menées en France et au Liban mais aussi en Afrique en réalisant la première enquête au Sénégal. Le sous-typage des isolats a été réalisé par PCR en temps réel en ciblant un domaine du gène de l’ARNr 18S suivi d’un séquençage direct du produit de PCR. Au Liban, la prévalence de Blastocystis était de 20% dans la population globale avec une corrélation entre le ST1 et le développement de symptômes gastro-intestinaux. Dans le même pays, cette prévalence dépassait les 60% chez des patients symptomatiques et des écoliers. Au Sénégal, la prévalence observée est la plus importante jamais décrite pour ce parasite puisqu’elle atteignait 100% dans une population d’une centaine d’enfants vivant en milieu rural. Ces données soulignent l’impact socioéconomique de la blastocystose dans les pays en développement où les conditions sanitaires sont souvent précaires. En France, une prévalence importante de 18% a pourtant été observée dans une large étude épidémiologique englobant des patients présentant ou non des symptômes et suivis dans 11 hôpitaux répartis sur tout le territoire français. Le ST3 est prédominant suivi des STs 1, 2 et 4 comme dans une majorité de pays à travers le monde. Le deuxième axe s’est concentré sur l’identification des facteurs de risque de transmission de Blastocystis à l’homme. Le parasite a été recherché dans les selles de vaches et de patients ainsi que dans des échantillons d’eau consommée par l’homme et les animaux dans une région géographique limitée du Nord Liban. 30% des échantillons humains, 69% des échantillons d'eau et 80% des échantillons de bovins étaient positifs pour le parasite. Le ST3 était prédominant dans les échantillons humains et d’eau suivi des ST1, ST2 et ST4. Par contre, ST10 et ST14 étaient prédominants chez les bovins mais ces deux STs n’ont pas été retrouvés dans les autres types d’échantillons. Pour expliquer l'absence des ST10 et ST14 dans ces échantillons, une transmission de ces STs par contact direct entre les bovins et/ou l'absence de formes kystiques transmissibles pour ces STs ont été proposées. Ce parasite a aussi été recherché dans les selles de nombreux groupes d’animaux du zoo de La Palmyre en France. Nous avons montré que près de 40% des selles analysés étaient positives pour Blastocystis et identifié de nouveaux réservoirs d'infections pour l’homme chez les carnivores. La prévalence du parasite atteignait 60% chez les primates chez lesquels les ST1 à ST5 identifiés sont identiques à ceux observés chez l'homme confirmant la faible spécificité d’hôte de ces STs. Dans une autre étude, la prévalence de Blastocystis était de seulement 3,5% dans une population de chiens en France suggérant que cet animal n'est pas un hôte naturel de Blastocystis. Enfin, pour clarifier la pathogénicité de ce parasite, le troisième axe de mes travaux a souligné le caractère invasif de Blastocystis dans un cas de péritonite appendiculaire chez une fillette de 9 ans de retour du Maroc. Seul Blastocystis a été détecté dans les selles, l’appendice, le liquide péritonéal et le sac de Douglas de cette patiente. Une gastro-entérite s’est de plus déclarée simultanément chez 26 membres de la famille de l'enfant suggérant une épidémie qui pourrait trouver son origine dans la consommation commune d’une eau contaminée<br>Blastocystis sp. is an anaerobic parasitic protozoa found in the digestive tract of humans and numerous animals. To date, it is the most common intestinal parasite found in human feces with worldwide distribution. Seventeen subtypes (ST1-ST17) have been described based on the comparison of SSU rRNA gene sequences. Blastocystis infection is associated with various gastrointestinal disorders and many studies suggest a correlation between Blastocystis STs and pathogenicity. My work was developed on three different topics. The first concerned the prevalence and the genetic biodiversity of the parasite in human populations. Epidemiological studies were conducted in France and Lebanon but also in Africa by performing the first survey of this parasite in Senegal. Subtyping of the isolates was performed by real-time PCR targeting a domain of the SSU rRNA gene followed by direct sequencing of the PCR product. In Lebanon, the prevalence of Blastocystis reached 20% in the general population and we demonstrated a correlation between ST1 infection and the presence of symptoms. In the same country, this prevalence was 60% in schoolchildren and patients presenting gastrointestinal symptoms. Strikingly, the prevalence of Blastocystis in a population of one hundred children living in a rural area reached 100% in Senegal and more than half of the infected children by the parasite presented gastrointestinal disorders. These latter studies highlighted the socioeconomic impact of blastocystosis in developing countries with poor hygiene sanitation. In France, a large-scale molecular epidemiological study was performed including patients presenting or not gastrointestinal symptoms. Stool samples were collected during winter and summer in 11 hospitals spread all over the French territory. We observed a high prevalence of Blastocystis in the french population with an average of 18.2% and the predominance of ST3 followed by ST1, ST4 and ST2 as in numerous countries. We also identified seasonal variations since the average prevalence of the parasite is 13.6% in winter and 23.1% in summer. The second topic focused on the identification of the risk factors of Blastocystis transmission to humans. We searched this parasite in bovid and human stools as well as in drinking water samples consumed by bovids and breeders in a limited geographic area of North-Lebanon. 30% of human samples, 69% of water samples and 80% of bovid samples were positive for the parasite. Interestingly ST3 is predominant in human and water samples followed by ST1, ST2 and ST4. ST10 and ST14 were predominant in bovid but both STs are lacking in human and water samples. To explain the lack of ST10 and ST14 in human and water samples, we suggested a transmission of these STs occurring through direct contact between bovid and / or the absence of transmissible cystic forms of these STs. Furthermore, this parasite was searched in the stools of numerous animal groups in the zoo of La Palmyre in France. We showed that nearly 40% of the analyzed stools were positive for Blastocystis and identified new reservoirs of human infections in carnivores. The prevalence of the parasite reached 60% in primates in which the identified ST1 to ST5 are identical to those observed in humans confirming the limited host specificity of these STs. In another study, we showed that the prevalence of Blastocystis was of only 3.5% in a population of one hundred dogs in France suggesting that this pet is not a natural host of Blastocystis. Finally, to clarify the pathogenicity of this parasite, the third topic highlighted the invasive character of Blastocystis observed in a case of appendicular peritonitis in a 9-year old girl returning from Morocco. Only Blastocystis was detected in stools, appendix, peritoneal liquid and Douglas pouch of the patient. Interestingly, simultaneous gastroenteritis occurred in 26 members of the child’s family suggested an outbreak with contaminated water as probable origin

Dans le cadre des interactions heterospecifiques durables, une etude des relations hote-parasite est realisee a partir du modele metacestode e. Multilocularis/hote intermediaire experimental (la gerbille de chine meriones unguiculatus). Dans un premier temps, les parametres influencant la dissemination de l'echinocoque (en tant que population clonale) sont etudies au travers de differents protocoles d'infestations secondaires. Les differents types de formes larvaires a l'origine des migrations dans l'hote sont decrits ainsi que les voies de migration preferentiellement empruntees. Une attention particuliere est portee a l'atteinte du systeme lymphatique. Les caracteristiques des infestations secondaires sous-cutanees et intra-peritoneales, sont discutees et comparees a celles de l'echinococcose primaire. Dans un deuxieme temps, les modes de differenciation du metacestode en tant qu'individu sont analyses grace a des etudes morphologiques et biochimiques. Les phases de l'ontogenese du parasite sont etablies en fonction des transformations tegumentaires. Les marquages des glycoconjugues a l'aide de lectines confirment la mise en evidence d'une regionalisation tegumentaire et d'une evolution des relations hote-parasite favorisant le developpement du metacestode. Une etude preliminaire des glycoproteines parasitaires est realisee a l'aide des techniques de sds-page et western blotting. Ces resultats suggerent que l'ontogenese du parasite serait determinee par des facteurs intrinseques et extrinseques (milieux physico-chimiques et immunologiques) evoluant dans le temps et l'espace

L'objectif de notre étude a été d'apporter des connaissances nouvelles sur la dynamique de la relation hôte-parasite dans le cas particulier du chevreuil et des nématodes qu'il héberge. Elle s'est déroulée sur 5 années et a permis la collecte et l'analyse coprologique puis statistique de 1607 prélèvements provenant de 7 populations de chevreuils contrastées sur le plan du milieu et de la densité relative aux ressources. L'helminthofaune observée correspond aux espèces de nématodes digestifs et pulmonaires généralement retrouvées en France. Après avoir établi la validité de la méthode coprologique dans le cadre d'une comparaison inter-populationnelle, nous avons mis en évidence la relation négative unissant la masse corporelle des chevreuils à leurs charges parasitaires digestives et pulmonaires. Nous avons également pu observer que la distribution des oeufs et des larves dans les matières fécales était agrégative. L'agrégation parasitaire est plus forte ou tend à l'etre chez les adultes, en particulier chez les femelles et lorsque l'abondance de la population d'hôtes est faible. Elle est plus faible ou tend à l'être chez les faons et les individus âgés de plus de 8 ans et lorsque les populations de chevreuils sont très abondantes par rapport aux ressources disponibles. Les prévalences et les indices parasitaires sont également plus élevés chez les faons et les chevreuils de plus de 8 ans que chez les adultes de 2 à 7 ans, chez les mâles que chez les femelles, en automne qu'en hiver, Lorsque les populations sont abondantes par rapport aux ressources et lorsque les chevreuils sont porteurs de Dictyocaulus noerneri. L'ensemble de nos résultats permet de penser que les parasites peuvent avoir un impact sur la dynamique des populations de chevreuils et en particulier un rôle régulateur. La construction d'un indicateur de la qualité de la relation entre les populations de chevreuils et leur milieu est proposée dans le contexte de la gestion de ces populations.

L'espece schistosoma intercalatum, agent de la schistosomose rectale en afrique, offre un double interet: fondamental par les questions non resolues que posent sa biologie et son ecologie, applique lie a la morbidite encore mal appreciee qu'elle cause chez l'homme. Les recherches experimentales de ce travail portent sur l'etude des interactions hote-parasite a la fois au niveau du compartiment mollusque et du compartiment vertebre, et des mecanismes de competition interspecifique dans les infestations mixtes s. Intercalatum-s. Mansoni. Il a ete demontre l'existence d'une compatibilite presque parfaite entre s. Intercalatum (souche de basse guinee) et differentes populations de b. Forskalii, originaires ou non de pays d'endemie. L'etude de la permissivite des rongeurs sauvages a montre que arvicanthis niloticus est un hote particulierement permissif vis-a-vis de s. Intercalatum, confirmant ainsi les hypotheses anciennes sur la participation possible des rongeurs dans le cycle de s. Intercalatum soit en qualite d'hotes reservoirs, soit en qualite d'hotes definitifs. L'etude des interactions entre s. Intercalatum et s. Mansoni a demontre l'existence de plusieurs mecanismes d'isolement reproductif prezygotique entre les deux especes. Ces mecanismes sont d'ordre immunologique et d'ordre sexuel. Sur le plan immunologique, il a ete mis en evidence qu'une primo-infestation par s. Mansoni immunise tres significativement le vertebre contre s. Intercalatum, ce qui contribue a limiter la rencontre entre les deux especes. Sur le plan sexuel, les infestations simultanees s. Intercalatum-s. Mansoni ont revele, pour la premiere fois chez les schistosomes, l'existence de mecanismes de reconnaissance favorisant l'appariement entre partenaires de la meme espece. La preference homospecifique d'accouplement est si forte qu'elle peut impliquer un changement de partenaire. Il a enfin ete demontre que les couples heterospecifiques engendrent par parthenogenese meiotique heploide une descendance sterile. Ce travail s'ouvre vers des applications epidemiologiques dans la mesure ou certains resultats experimentaux sont de nature a expliquer les mecanismes s'opposant a l'extension geographique de s. Intercalatum.

The diplomonads are a diverse group of eukaryotic flagellates found in microaerophilic and anaerobic environments. The most studied diplomonad is the intestinal parasite Giardia intestinalis, which infects a variety of mammals and cause diarrheal disease. Less is known about Spironucleus salmonicida, a parasite of salmonid fish, known to cause systemic infections with high mortality. We created a transfection system for S. salmonicida to study cellular functions and virulence in detail (Paper I). The system was applied to explore the mitochondrion-related organelle (MRO) in S. salmonicida. We showed that S. salmonicida possesses a hydrogenosome (Paper II) with a higher metabolic capacity than the corresponding MRO of Giardia, the mitosome. Evolutionary analysis of key hydrogenosomal proteins showed ancient origin, indicating their presence in the ancestral diplomonad and subsequent loss in Giardia. Annexins are of evolutionary interest since these proteins are found across all kingdoms. Annexin-like proteins are intriguingly expanded into multigene families in Giardia and Spironucleus. The annexins of S. salmonicida were characterized (Paper III) with distinct localizations to various cellular structures, including a putative adhesion structure anterior in the cell. The disease-causing Giardia trophozoites differentiate into infectious cysts, a process essential for transmission and virulence of the parasite. Cysts are often spread via contaminated water and exposed to environmental stressors, such as UV irradiation. We studied the survival and transcriptional response to this stress factor (Paper IV) and results showed the importance of active DNA replication machinery for parasite survival after DNA damage. In addition, we studied transcriptional changes along the trajectory of encystation (Paper V), which revealed a coordinated cascade of gene regulation. This was observed for the entire transcriptome as well as putative regulators. Large transcriptional changes appeared late in the process with the majority of differentially regulated genes encoding hypothetical proteins. We studied the localizations of several of these to gain information of their possible function. To conclude, the diplomonads are complex eukaryotic microbes with cellular processes adjusted to match their life styles. The work in this thesis has provided insight of their adaptations, differences and similarities, but also new interesting leads for future studies of diplomonad biology and virulence.

The development of molecular tools for detection and typing of unicellular intestinal parasites has revealed genetic diversities in species that were previously considered as distinct entities. Of great importance is the genetic distinction found between the pathogenic Entamoeba histolytica and the non-pathogenic Entamoeba dispar, two morphologically indistinguishable species. Blastocystis sp. is a ubiquitous intestinal parasite with unsettled pathogenicity. Molecular studies of Blastocystis sp. have identified 17 genetic subtypes, named ST1-17. Genetically, these subtypes could be considered as different species, but it is largely unknown what phenotypic or pathogenic differences exist between them. This thesis explores molecular methods for detection and genetic subtyping of unicellular intestinal parasites, with special focus on Blastocystis. We found that PCR-based methods were highly sensitive for detection of unicellular intestinal parasites, but could be partially or completely inhibited by substances present in faeces. A sample transport medium containing guanidinium thiocyanate was shown to limit the occurrence of PCR inhibition. The prevalence of Blastocystis in Swedish university students was over 40%, which is markedly higher than what was previously estimated. Blastocystis ST3 and ST4 were the two most commonly found Blastocystis subtypes in Sweden, which is similar to results from other European countries. Blastocystis sp. and Giardia intestinalis were both commonly detected in Zanzibar, Tanzania, each with a prevalence exceeding 50%. Blastocystis ST1, ST2, and ST3 were common, but ST4 was absent. While G. intestinalis was most common in the ages 2-5 years, the prevalence of Blastocystis increased with increasing age, at least up to young adulthood. We found no statistical association between diarrhoea and Blastocystis sp., specific Blastocystis subtype or G. intestinalis. Metagenomic sequencing of faecal samples from Swedes revealed that Blastocystis was associated with high intestinal bacterial genus richness, possibly signifying gastrointestinal health. Blastocystis was also positively associated with the bacterial genera Sporolactobacillus and Candidatus Carsonella, and negatively associated with the genus Bacteroides. Blastocystis ST4 was shown to have limited intra-subtype genetic diversity and limited geographic spread. ST4 was also found to be the major driver behind the positive association between Blastocystis and bacterial genus richness and the negative association with Bacteroides.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2017-04-03T13:51:17Z No. of bitstreams: 1 elisabethcamposdeandrade.pdf: 1614173 bytes, checksum: 02bbfb073bbecc90f3e7f67d2c386c90 (MD5)<br>Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-04-03T19:01:36Z (GMT) No. of bitstreams: 1 elisabethcamposdeandrade.pdf: 1614173 bytes, checksum: 02bbfb073bbecc90f3e7f67d2c386c90 (MD5)<br>Made available in DSpace on 2017-04-03T19:01:36Z (GMT). No. of bitstreams: 1 elisabethcamposdeandrade.pdf: 1614173 bytes, checksum: 02bbfb073bbecc90f3e7f67d2c386c90 (MD5) Previous issue date: 2009-03-26<br>FAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas Gerais<br>As parasitoses intestinais são um importante problema de saúde pública principalmente nos países subdesenvolvidos ou em desenvolvimento. Embora o parasitismo intestinal seja amplamente reconhecido como relevante no contexto epidemiológico de diversas comunidades, os estudos sobre o assunto são ainda insuficientes, principalmente no Brasil. Em vista da dificuldade de diagnóstico específico das parasitoses, muitas vezes são realizados tratamentos empíricos com mais de uma droga. O objetivo principal do presente estudo foi avaliar a efetividade e segurança do uso de nitazoxanida no tratamento de poliparasitoses comparado à terapêutica tradicional ofertada pelo serviço público. Além deste objetivo, foi possível investigar a prevalência e os fatores associados às parasitoses intestinais na população de Colônia do Paiol, uma comunidade quilombola, na Zona da Mata, Minas Gerais. Na comunidade analisada, procedeuse um estudo transversal por censo, sendo que dos 425 moradores, 391 (92%) foram avaliados através de questionário estruturado e exame coproparasitológico. Os resultados mostraram uma alta positividade (63,8%), sendo as espécies patogênicas mais freqüentes Ascaris lumbricoides (22,4%) e Trichuris trichiura (17,9%). O poliparasitismo ocorreu em 36,5% dos investigados. O ensaio clínico, controlado, duplo cego, randomizado avaliou 65 indivíduos em dois grupos de tratamento. A taxa de cura foi de 32,4% e 38,7% com a nitazoxanida e com o tratamento convencional, respectivamente, mas esta diferença não foi estatisticamente significativa (p=0,599). A ocorrência de vômito (p= 0,031) esteve associada ao tratamento convencional e de urina esverdeada ao uso de nitazoxanida (p=0,002). Os outros efeitos adversos foram independentes da droga. Houve diferença estatisticamente significativa em relação à cor da pele e a taxa de cura para ambos os tratamentos. A menor eficácia efetividade foi apresentada pelos indivíduos de cor preta. São necessários outros estudos para esclarecer a baixa efetividade nos casos de poliparasitismo, assim como, reavaliar as práticas preventivas e terapêuticas, com o uso de novas drogas e de agentes de largo espectro, podendo a nitazoxanida ser uma droga alternativa neste contexto. Agrega-se às novas possibilidades terapêuticas, a necessidade de políticas públicas que garantam qualidade de vida, através de saneamento básico, educação para saúde e acesso ao sistema público de saúde, minimizando as iniqüidades na sociedade.<br>Intestinal parasitism is an important public health concern, chiefly in underdeveloped or developing countries. Although widely recognized as a relevant community epidemiological issue, intestinal parasitism has not been sufficiently studied in Brazil. Because specific diagnosis is difficult and generally cumbersome, empiric treatment, sometimes with more than one drug, is frequently employed. The main aim of this study was the assessment of the effectiveness and safety of nitazoxanide for the treatment of intestinal polyparasitism, as compared to traditional therapy provided by the public service. The study also investigated the prevalence of and factors associated with intestinal parasitism in the population of Colônia do Paiol, a quilombola community from the Zona da Mata, Minas Gerais, Brazil. A census-based cross-sectional study used a structured questionnaire and stool examination to assess 391 people (92%) of the 425 inhabitants of the community. The frequency of intestinal parasitism was as high as 63.8%, with predominance of Ascaris lumbricoides (22.4%) and Trichuris trichiura (17.9%). Polyparasitism occurred in 36.5% of those investigated. A double-blind randomized controlled trial assessed 65 individuals in two treatment groups. Cure rates were 32.4% and 38.7% with nitazoxanide and conventional treatment, respectively, but the difference was not statistically significant (p = 0.599). Vomiting (p = 0.031) was associated with conventional treatment and greenish urine with nitazoxanide use (p = 0.002). Other untoward effects were independent of which drug was used. There was a statistically significant difference concerning skin color and cure rates for both treatments. Dark-skinned subjects had lower cure rates. Further studies are necessary to clarify the reasons for the low effectiveness found in these cases of polyparasitism, and to reevaluate preventive and therapeutic approaches with new and broad-spectrum drugs, nitazoxanide being an option in this context. In addition to new therapeutic approaches, there is a clear need to develop public policies that, through the provision of basic sanitation, health education and access to the public health system, assure quality of life and minimize inequity.

Les conditions climatiques ont un role preponderant sur la repartition geographique et temporelle des differentes especes de nematodes parasites des ruminants. Sur un exemple tire du parasitisme des bovins du centre-ouest d'argentine, temperature et pluviometrie contribuent a expliquer les niveaux d'infestation par les principaux genres. Un travail plus experimental est developpe en vue d'estimer le role de l'eau a proximite immediate des ufs et jeunes larves des nematodes (dans les feces), avec ou sans celui de la temperature, sur la partie du cycle de developpement allant de l'uf a la larve infestante. Par des depots de feces d'animaux parasites dans des situations climatiques diversifiees, il est montre que la temperature au sein des bouses etait le premier facteur conditionnant le taux de developpement des especes parasitant les bovins et que c'etait l'humidite ponderale des feces dans le cas des parasites d'ovin. Les exigences thermique et surtout hydriques sont definies au laboratoire pour huit especes parasites de bovins, ovins et cervides multipliees sur ovin. L'adaptation aux faibles humidites etait le principal caractere specifique. La taille des larves infestantes s'est averee dependre des conditions de developpement. Par des tests de laboratoire, il est montre que les petites larves developpees a des humidites faibles migraient et se degainaient plus lentement que les normales et semblaient avoir plus de mal a s'etablir chez l'hote. L'adaptation genetique aux conditions seches d'une lignee a ete tentee par une selection pendant 5 generations mais n'a pas revele d'evolution du taux de developpement dans ces conditions