Orientador: João Aristeu da Rosa / Banca: Marlene Tiduko Ueta / Banca: Elenice Messias do Nascimento Gonçalves / Banca: Silmara Marques Allegretti / Banca: Mara Cristina Pinto / Resumo: A estrongiloidíase é uma doença parasitária que tem distribuição mundial heterogênea, cuja prevalência é dividida em três categorias: esporádica (<1%), endêmica (1-5%) e hiperendêmica (>5%). Na região de Araraquara, estudos de frequência de enteroparasitoses vêm sendo conduzidos desde 1970 e mostram que a estrongiloidíase é endêmica na região. Com o intuito de realizar caracterização morfométrica de diferentes isolados humanos desse helminto, provenientes dos municípios de Araraquara, Américo Brasiliense, Gavião Peixoto, Motuca e Rincão foi feita a mensuração de larvas rabditóides e filarióides e sequenciamento da região ITS-1 do rDNA das larvas filarióides. Com o auxilio do programa de captura e mensuração de imagens Motic Images Advanced 3.2., os parâmetros das larvas rabditóides e filarióides propostos por TEIXEIRA, 1996, foram mensurados após fixação das mesmas em TAF. A extração do DNA genômico foi realizada segundo protocolo descrito por NILFOROUSHAN et al., 2007 e em seguida realizou-se a amplificação do ITS-1 por meio de NESTED-PCR. Os fragmentos de aproximadamente 680 pb obtidos foram purificados, sequenciados e a comparação de sequências foi realizada pelo programa ClustalW. Apenas com os resultados da análise morfométrica, não foi possível caracterizar diferentes cepas de S. stercoralis, uma vez que... (REsumo completo, clicar acesso eletrônico abaixo) / Abstract: Strongyloidiasis is a parasitic disease that occurs worldwide heterogeneous, whose prevalence is divided into three categories: sporadic (<1%), endemic (1-5%) and hyperendemic (> 5%). In the region of Araraquara, studies the frequency of intestinal parasites have been conducted since 1970 and show that strongyloidiasis is endemic in the region. In order to perform morphometric characterization of different isolates of human intestinal worms, from the municipalities of Araraquara, Américo Brasiliense, Gavião Peixoto, Motuca and Rincão the measurement was made of rhabditiforms and filariforms larvae and sequencing of the ITS-1 rDNA filariform larvae. With the help of the program capture and measurement of images Motic Images Advanced 3.2., the rhabditiforms and filariforms larvae parameters proposed by TEIXEIRA, 1997, were measured after fixed on the TAF. The extraction of genomic DNA was performed according to protocol described by NILFOROUSHAN et al., 2007 and then held the amplification of ITS-1 by nested-PCR. Fragments of about 680 bp obtained were purified, sequenced and the sequence comparison was performed by the program ClustalW. Only the results of the morphometric analysis was not possible to characterize different S. stercoralis strains, since the values of the parameters measured were quite homogeneous in larvae... (Complete abstract click electronic access below) / Doutor

The study was aimed to obtain basic information about the distribution of intestinal parasites of mammals and birds in Svalbard. Faeces of different species were used to detect intestinal parasites. All samples were collected during two seasons and examined by microscopic methods and molecular diagnostic was used for detection of microsporidia, cryptosporidia, coccidia and giardiae.

Made available in DSpace on 2014-06-11T19:33:23Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-11-25Bitstream added on 2014-06-13T18:07:29Z : No. of bitstreams: 1 mine_jc_dr_arafcf.pdf: 1238485 bytes, checksum: c0cab12233278cce39285c34cc901f95 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A estrongiloidíase é uma doença parasitária que tem distribuição mundial heterogênea, cuja prevalência é dividida em três categorias: esporádica (<1%), endêmica (1-5%) e hiperendêmica (>5%). Na região de Araraquara, estudos de frequência de enteroparasitoses vêm sendo conduzidos desde 1970 e mostram que a estrongiloidíase é endêmica na região. Com o intuito de realizar caracterização morfométrica de diferentes isolados humanos desse helminto, provenientes dos municípios de Araraquara, Américo Brasiliense, Gavião Peixoto, Motuca e Rincão foi feita a mensuração de larvas rabditóides e filarióides e sequenciamento da região ITS-1 do rDNA das larvas filarióides. Com o auxilio do programa de captura e mensuração de imagens Motic Images Advanced 3.2., os parâmetros das larvas rabditóides e filarióides propostos por TEIXEIRA, 1996, foram mensurados após fixação das mesmas em TAF. A extração do DNA genômico foi realizada segundo protocolo descrito por NILFOROUSHAN et al., 2007 e em seguida realizou-se a amplificação do ITS-1 por meio de NESTED-PCR. Os fragmentos de aproximadamente 680 pb obtidos foram purificados, sequenciados e a comparação de sequências foi realizada pelo programa ClustalW. Apenas com os resultados da análise morfométrica, não foi possível caracterizar diferentes cepas de S. stercoralis, uma vez que... / Strongyloidiasis is a parasitic disease that occurs worldwide heterogeneous, whose prevalence is divided into three categories: sporadic (<1%), endemic (1-5%) and hyperendemic (> 5%). In the region of Araraquara, studies the frequency of intestinal parasites have been conducted since 1970 and show that strongyloidiasis is endemic in the region. In order to perform morphometric characterization of different isolates of human intestinal worms, from the municipalities of Araraquara, Américo Brasiliense, Gavião Peixoto, Motuca and Rincão the measurement was made of rhabditiforms and filariforms larvae and sequencing of the ITS-1 rDNA filariform larvae. With the help of the program capture and measurement of images Motic Images Advanced 3.2., the rhabditiforms and filariforms larvae parameters proposed by TEIXEIRA, 1997, were measured after fixed on the TAF. The extraction of genomic DNA was performed according to protocol described by NILFOROUSHAN et al., 2007 and then held the amplification of ITS-1 by nested-PCR. Fragments of about 680 bp obtained were purified, sequenced and the sequence comparison was performed by the program ClustalW. Only the results of the morphometric analysis was not possible to characterize different S. stercoralis strains, since the values of the parameters measured were quite homogeneous in larvae... (Complete abstract click electronic access below)

The number of tissue eosinophils were counted and phospholiphase B activity was assayed in the intestines of nonsensitized and sensitized and sensitized mice infected with Trichinella spiralis.

Orientador: João Aristeu da Rosa / Banca: Vera Lucy de Santi Alvarenga / Banca: Vera Lucia Pagliusi Castilho / Resumo: 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. / Abstract: 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. / Mestre

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) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Universidade Estadual Paulista (UNESP) / 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... / 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.

Pooled faecal specimens of pigs were collected from the floor on two farms (A and B) and parasitologically examined. Investigations were divided into the five period: winter 2005 (A), spring 2005 (A), autumn 2005 (B), winter 2006 (B) and spring 2006 (B). In total, we collected 249 faecal specimens of pigs, of them 91 specimens were collected from farm A and 158 specimens from farm B. In farm A we collected 32 specimens from post-weaned piglets and in farm B 20 specimens from post-weaned piglets. Piglets were housed on litter stabling on both farms.

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. / 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.

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. / Master of Science

In total, 76 birds, mainly breeding females, were captured in years 2007-2009. Birds were inspected for the presence of avian influenza (AI) viruses and the AI antibodies. They were inspected as well for the presence of bacteria Borrelia burgdorferi s. l. and Anaplasma phagocytophilum and protozoan Leucocytozoon simondi in blood. The feces were examined for the oocysts and eggs of intestinal parasites. AI viruses were found in 4 cases, while over 80% of inspected birds had AI antibodies. Bacteria B. burgdorferi s. l. and A. phagocytophilum were found in 2 and 1 case respectively. Protozoan L. simondi was not found in any blood smear. The oocysts from protozoan genera Eimeria and Tyzzeria and eggs from roundworm genera Capillaria, Amidostomum and Echinuria were observed in feces. Statistical analyses were due to the small size of our data set inconclusive.