Dissertations / Theses on the topic 'Hominis'

The author challenges the terminology «legal presumptions» and «judicial presumptions», and rather refers to presumptions established by rules of presumption and to hominis presumptions. He argues that the best way to differentiate between them is by showing the contrast between «it shall be presumed» (syntagm proper to practical reasoning) and «it is presumable» (syntagm proper to theoretical reasoning). The text clarifies the relationship between the so-called hominis presumptions and the factual inferences or evidential inferences, in general. He answers the question of what the «it is presumed» syntagm (proper to the hominis presumptions) brings with respect to the «it is probable» syntagm (proper of all evidentiary inferences).
El autor cuestiona la terminología «presunciones legales» y «presunciones judiciales» y, más bien, se refiere a las presunciones establecidas por normas de presunción y a las presunciones hominis. Defiende que la mejor manera de diferenciar unas de otras es mostrando la distancia que media entre «debe presumirse» (sintagma propio del razonamiento práctico) y «es presumible» (sintagma propio del razonamiento teórico). El texto aclara las relaciones entre las llamadas presunciones hominis y las inferencias fácticas o inferencias probatorias, en general, respondiendo a la pregunta sobre qué aporta el sintagma «es presumible» (propio de las presunciones hominis) frente al sintagma «es probable» (propio de todas las inferencias probatorias).

En los últimos años diversos trabajos ponen de manifiesto la existencia de variaciones intraespecíficas entre aislados de Blastocystis hominis. La descripción de diferentes perfiles proteicos, cariotipos y zimodemas constituyen pruebas de la posible existencia de poblaciones morfológicamente idénticas, que quizás estén dotadas de un potencial patogénico diferente. El objeto de este estudio es el de establecer asociaciones entre variantes genéticas y enzimáticas y distinta significación clínica, para lo cual se realizaron los siguientes procedimientos:(1) Obtener aislados clínicos de pacientes sintomáticos y asintomáticos, cultivarlos y axenizarlos.(2) Comprobar el grado de variación genética críptica en aislados de B. hominis, mediante el análisis de polimorfismo de fragmentos de restricción a partir de dos amplicones diferentes.(3) Evaluar el poder discriminatorio de secuencias repetitivas de ADN como marcadores de diversidad.(4) Demostrar la existencia de poblaciones isoenzimáticas. Se analizaron un total de 260 muestras fecales, de entre las remitidas al Servicio de Microbiología del Hospital Clínico Universitario de Valencia; que fueron sometidas al siguiente protocolo: exámen parasitológico para la identificación de B. hominis y de otros posibles parásitos; y un coprocultivo con el objeto de determinar la existencia de enteropatógenos bacterianos. Se observó B. hominis en 56 de estas muestras, lo que supuso una prevalencia del 21,5%; y se procedió a su cultivo en medio BDM para su posterior axenización, todo ello de acuerdo con el protocolo descrito por Lanuza et al. (1997). Se consiguió la adaptación al cultivo de 38 aislados, de los que se consiguió axenizar 18. Se incluyeron en el estudio 13 cultivos puros procedentes de colección, por lo que finalmente se analizaron un total de 51 aislados: 31 axénicos y 20 monoxénicos. Una vez obtenidos los aislados se llevaron a cabo los ensayos de "riboprinting" (Böhm-Gloning,1997; Clark, 1997), consistentes en amplificar la secuencia que codifica el ARN de la subunidad menor ribosomal y analizar la longitud de los fragmentos obtenidos tras restricción enzimática de dicha región. Extraído el ADN de acuerdo con el procedimiento descrito por Clark (1997) consistente en una extracción fenólica; la amplificación se realizó en dos reacciones distintas: la primera con los cebadores RD3 y RD5 (Clark, 1997) que amplifican la totalidad del gen con lo que se obtiene un amplicón de 2.540 pb; y la segunda con F1 y R1 (Böhm-Gloning et al., 1997) que amplifican sólo una parte del gen, por lo que se obtiene un producto de 1.100 pb. Los productos de amplificación fueron digeridos separadamente con las endonucleasas Rsa I, Hinf I y Alu I. Para el análisis se transformaron las distancias de migración en tamaños relativos al compararlos con marcadores de peso molecular, utilizando el programa informático 1DManager. El criterio establecido para diferenciar entre patrones de RFLP fue considerar que eran distintos los que diferían en uno o en más fragmentos. Analizados los perfiles obtenidos para cada aislado, se obtuvieron los índices de homología entre cepas aplicando el coeficiente de Dice. Los resultados obtenidos tras el análisis conjunto de todos los patrones de bandas fue la obtención de 5 ribodemas (R1 a R5), en los que se valoró si existía relación estadística entre la pertenencia a un ribodema y padecer diarrea aguda o crónica, por lo que se analizaron los datos aplicando el test CHI-CUADRADO y se definieron el riesgo relativo (RR) y el coeficiente de correlación de Pearson (p).Los resultados demostraron una asociación estadísticamente significativa entre diarrea aguda e inclusión en el ribodema R2. Lo cual nos confirmó que la especie B. hominis es genéticamente heterogénea, y que existían diferentes poblaciones con diferencias en la capacidad patogénica. En el estudio también se valoró la eficacia de secuencias repetitivas de ADN como marcadores de diversidad, para lo cual se utilizaron los cebadores TR7 y TR8 (Init et al., 1999), demostrando que se podían establecer diferencias intraespecíficas a partir de un fragmento mayoritario, es decir el que más veces se repetía, considerando que eran cepas pertenecientes al mismo patrón las que coincidían en dicho fragmento. Los resultados demostraron que esta técnica es únicamente válida en la detección de variación intraespecífica en cultivos puros, ya que los cebadores no discriminan suficientemente cuando en la muestra existe ADN bacteriano además del de B. hominis. No se encontraron asociaciones estadísticas entre la pertenencia a alguno de los patrones definidos y la presentación de un proceso diarreico determinado. Finalmente el análisis de las actividades enzimáticas: Glucosa-6-fosfato deshidrogenasa, malato deshidrogenasa, enzima málico, hexoquinasa, fosfoglucomutasa, glucosa fosfato isomerasa, glutámico-oxalacético transaminasa y fosfogluconato deshidrogenasa; consistió en la extracción de las proteínas de los cultivos puros, su separación en geles de poliacrilamida, y el consiguiente revelado utilizando sustancias tamponadas, con los sutratos, cofactores y coenzimas necesarios para cada enzima, de acuerdo con el protocolo de Selander et al. (1985).Todos los aislados presentaron todas las actividades enzimáticas y compartieron idéntico patrón en todos los enzimas a excepción de la malato deshidrogenasa; cuyas diferencias en la movilidad electroforética de una de sus bandas, permitió definir tres poblaciones isoenzimáticas. No se encontraron asociaciones estadísticas entre los zimodemas definidos y la presentación de un determinado proceso diarreico. Los resultados permitieron extraer las siguientes conclusiones:1ª.- Todos los métodos moleculares empleados demostraron, aunque con desigual capacidad de discriminación, que B. hominis es un organismo con marcada heterogeneidad genética.2ª.- El análisis de la secuencia del ARN ribosomal 16S, mediante técnicas de RFLP, ha detectado variantes asociadas a determinados estados mórbidos. El protocolo que utiliza los cebadores RD3 y RD5 es el que ha mostrado un mayor poder de resolución.3ª.- Consideramos que para poder comparar datos de RFLP de forma efectiva es necesaria la utilización de protocolos normalizados, con cepas de referencia, un panel común de enzimas de restricción y una nomenclatura consensuada.4ª.- El estudio de secuencias repetitivas utilizando TR7 y TR8 resulta eficaz en la detección de variación intraespecífica únicamente en aislados axénicos, lo cual limita su utilización como marcador de diversidad.5ª.- Las diferencias encontradas en la movilidad del enzima malato deshidrogenasa demostraron la existencia de tres poblaciones isoenzimáticas, son que ninguna de ellas se asociara con un proceso determinado.6ª.- La homología encontrada entre nuestros aislados y los genotipos de origen humano y animal descritos en otras áreas geográficas sustentan la idea del origen zoonótico de la infección y demuestran la ausencia de variaciones genéticas regionales. Por lo que este trabajo concluye que B. hominis es una especie genéticamente heterogénea, y la asociación estadística observada confirma además la existencia de poblaciones con diferente patogenicidad.
Blastocystis hominis is the most prevalent specie of intestinal protozoa found in men of uncertain role in human disease. This study was undertaken to examine the degree of criptic genetic variation within B. hominis detectable using riboprinting (restriction fragment length polymorphism analysis of polymerase chain reaction amplified small subunit rDNA); the analysis of multiple target sequences by a single set of polymerase chain reaction primers; and the existence of demes with different pathogenic potential after to analyse the enzyme activity. The small subunit ribosomal RNA genes of 51 isolates were amplified using standard polymerase chain reaction conditions and the primers RD5 y RD3 (Clark, CG; 1992). The amplification products were digested with 3 restriction enzymes (Hinf I, Rsa I, Alu I). We used a similar method by the primers F1 y R1 (Böhm-Gloning et al., 1997) and the same restriction endonuclease enzymes. Extensive sequence variation was discovered in B. hominis ribosomal RNA genes and the isolates grouped of at least five differents patterns or ribodemes. The ribodeme R2 was the most frequently isolated variant, is composed of isolates obtained from patients with acute diarrhea. The statistical association found between R2 and acute diarrhea in the absence of other enteropathogens suggest the pathogenic role of this ribodema. The results based on the amplification products obtaines by a single set of polymerase chain reaction primers TR7 y TR8 (Init et al., 1999) showed intrastrain variations among isolates, but this set of PCR primers only can be used in the detection of variable DNA repeat patterns in axenic cultures because the reaction is not selectivity for lower eukayotes, it is present in prokaryotes too . The enzyme polymorphism of this organism has been demostrated by enzyme electrophoresis of 31 axenic B. hominis isolates by electrophoresis on polyacrylamide gel under nondenaturating conditions (PAGE). Eight enzyme systems were studied: GOT (EC 2.6.1.1), GPI (EC 5.3.1.9), EM (EC 1.1.1.40), G6PDH (EC 1.1.1.49), 6PGDH (EC 1.1.1.44), MDH (EC 1.1.1.37), HK (EC 2.7.1.1), PGM (EC 2.7.5.1. ). The isolates showed an identical isoenzyme profile by the seven activities, in the case of MDH the presence of one band with different electrophoretic migrations detected the existence the two zimodemes. The description of different zymodemes confirms the heterogeneity of this organism. This study illustrates the need to reexamine the role of B. hominis in disease taking the genetic diversity of the parasite into account.

Les mycoplasmes sont des bactéries à petit génome dérivées d’ancêtres à Gram positif par une succession de pertes de matériel génétique. Il a longtemps été considéré que la réduction génétique était la seule force régissant l’évolution de ces bactéries, cependant, des transferts horizontaux de grandes régions chromosomiques au sein et entre les espèces de mycoplasmes ont été récemment mis en évidence. Des éléments conjugatifs et intégratifs (ICE) découverts chez certaines espèces de mycoplasme pourraient être à l’origine de ces transferts. Ces ICEs codent les systèmes nécessaires pour leur excision, leur transfert conjugatif et leur intégration dans la cellule receveuse.Mycoplasma hominis est un mycoplasme commensal des voies génitales qui peut être responsable d’infections gynécologiques, d’infections néonatales et d’infections extragénitales. L’analyse préliminaire de génomes de M. hominis avait montré la présence de régions codantes caractéristiques des ICEs. Les objectifs de cette thèse étaient de rechercher et caractériser les ICEs chez 12 isolats cliniques de M. hominis entièrement séquencés et de déterminer la prévalence de ces ICEs au sein de l’espèce M. hominis. Pour cela, une étude rétrospective sur une période de 6 ans a été menée sur des isolats cliniques obtenus au CHU de Bordeaux. Les concentrations minimales inhibitrices des tétracyclines et des fluoroquinolones ainsi que les mécanismes de résistance ont été déterminés, permettant de disposer d’une collection d’isolats cliniques caractérisés pour l’étude des ICEs.Des ICEs de près de 30 kpb ont été trouvés en une ou plusieurs copies dans sept des 12 souches de M. hominis séquencées. Seulement cinq de ces ICEs semblaient fonctionnels puisqu’une forme circulaire a pu être détectée. Tous les ICEs de M. hominis présentaient une structure similaire avec un module spécifique de M. hominis d’environ 4-kpb, codant des protéines ayant des caractéristiques structurelles similaires à des effecteurs TAL (transcription activator-like), impliqués dans la reconnaissance de nucléotides et dans la transduction de signaux chez les bactéries symbiotiques. La caractérisation des mécanismes de résistance aux antibiotiques des isolats cliniques de M. hominis collectés au CHU de Bordeaux nous a permis de disposer d’une collection de 183 isolats isolés entre 2010 et 2015, parmi lesquels 14,8% étaient porteur du gène tet(M) responsable de la résistance aux tétracyclines, 2,7% étaient résistant à la lévofloxacine et 1,6% étaient résistants à la moxifloxacine par mutation des gènes de la topoisomérase IV et de l’ADN gyrase. Le screening de 120 de ces isolats cliniques a révélé une prévalence élevée des ICEs dans l’espèce M. hominis, mesurée à 45%. Il n’y avait pas de prédominance des ICEs dans les isolats portant le gène tet(M), suggérant que les ICEs n’étaient pas responsables de la dissémination de la résistance à la tétracycline.Des expériences complémentaires de conjugaison seront nécessaires pour confirmer la fonctionnalité des ICEs retrouvés dans l’espèce M. hominis. Cependant, la forte prévalence et le caractère très conservé des ICEs chez M. hominis suggèrent que ces ICEs pourraient conférer un avantage sélectif pour la physiologie ou la physiopathologie de la bactérie. Ce travail ouvre ainsi la voie à de futures études qui permettront une meilleure compréhension des transferts horizontaux de gènes et des facteurs de virulence chez M. hominis
Mycoplasmas are small-genome bacteria derived from Gram-positive ancestors by a succession of genetic material losses. It has long been considered that genetic reduction was the only force governing the evolution of these bacteria, however, horizontal transfers of large chromosomal regions within and between mycoplasma species have recently been reported. Conjugative and integrative elements (ICE) found in some species of mycoplasma may be responsible for these transfers. These ICEs encode the systems necessary for excision, conjugative transfer and integration into a recipient cell.Mycoplasma hominis is a commensal genital mycoplasma that can be responsible for gynecological infections, neonatal infections and extragenital infections. Preliminary analysis of M. hominis genomes had showed the presence of coding regions characteristic of ICEs. The objectives of this thesis were to search for and characterize ICEs in one reference strain and 11 fully sequenced M. hominis clinical isolates and to determine the prevalence of these ICEs in the M. hominis species. To do so, a retrospective study over a period of 6 years was conducted on clinical isolates collected at the Bordeaux University Hospital. The minimum inhibitory concentrations of tetracyclines and fluoroquinolones as well as resistance mechanisms were determined, providing a collection of clinical isolates characterized for the study of ICEs.ICEs of 27-30 kpb were found in one or two copies in seven of the 12 M. hominis sequenced strains. Only five of these ICEs seemed functional since circular forms of extrachromosomal ICE were detected. All M. hominis ICEs exhibited a similar structure consisting of a 4.0-5.1 kb module composed of five to six juxtaposed CDSs, encoding proteins that share common structural features with transcription activator-like (TAL) effectors, involved in polynucleotide recognition and signal transduction in symbiotic bacteria. The characterization of antibiotic resistance mechanisms in M. hominis clinical isolates collected at Bordeaux University Hospital enabled us to obtain a collection of 183 isolates isolated between 2010 and 2015, of which 14.8% harbored the tet(M) gene responsible for tetracycline resistance, 2.7% were resistant to levofloxacin and 1.6% were resistant to moxifloxacin by mutation in topoisomerase IV and DNA gyrase genes. Screening of 120 of these clinical isolates revealed a high prevalence of ICEs in M. hominis, measured to be 45%. The proportion of ICEs was not higher in isolates carrying the tet (M) gene, suggesting that ICEs were not responsible for the spread of tetracycline resistance.Additional mating experiments will be necessary to confirm the functionality of the ICEs found in the M. hominis species. However, the conserved and specific structure of M. hominis ICEs and the high prevalence in clinical strains suggest that these ICEs may confer a selective advantage for the physiology or pathogenicity of the bacteria. This work opens the way for future studies that will provide a better understanding of horizontal gene transfers and virulence factors in M. hominis

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CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico
Mycoplasmas are microorganisms lacking cell walls with reduced ability Biosintética, which makes them extremely fastidious growth. The strength of these microorganisms to antimicrobials used in his therapy as tetracyclines, macrolides and quinolones have been reported with increased frequency. The determination of antimicrobial susceptibility is particularly difficult because not shown in broth turbidity which complicates the standardization of the inoculum and are extremely susceptible pH conditions. The method most commonly used for this purpose is based on metabolic inhibition using specific substrates. This study evaluated the susceptibility of isolates of Mycoplasma hominis and Ureaplasma sp. stored in the period from 2011 to 2012. The methodology used for the isolation comprised culture techniques, for storage and freezing of the strains we used selective broth enriched PPLO and BHI, Thawing of isolates was succeeded subculture in broth enriched selective and differential agar A7 for phenotypic characterization. Quantification, identification and antimicrobial susceptibility testing was performed using the the triage system Mycofast Evolution ® Screening (Elitech Group), whose identification is based on bacterial susceptibility by Identibiotiqué system. The susceptibility of the isolates was determined against antimicrobial agents Doxiciclina 8 μg/ml, Roxitromicina 4 μg/ml and Ofloxacina 4 μg/ml, widely used in the empirical treatment of patients with urogenital tract infections. Our study showed high level of resistance of the isolates to doxicycline and ofloxacin. This is the first study involving isolation and susceptibility profile of these agents undertaken in the Amazonia.
Micoplasmas são microrganismos desprovidos de parede celular, com reduzida capacidade Biossintética, o que os torna extremamente fastidiosos ao crescimento. A resistência destes microrganismos aos agentes antimicrobianos utilizados na sua terapia como as tetraciclinas, macrolídeos e quinolonas tem sido relatadas com frequência cada vez maior. A determinação da susceptibilidade a antimicrobianos é particularmente difícil porque não mostram turbidez em caldo o que dificulta a padronização do inóculo e são extremamente susceptíveis as condições de pH. A metodologia utilizada para este fim baseia-se na inibição metabólica utilizando substratos específicos. O presente trabalho avaliou a susceptibilidade de isolados de Mycoplasma hominis e Ureaplasma sp. arrmazenados no período de 2011 a 2012. A metodologia utilizada englobou técnicas de cultura; para a estocagem e congelamento das cepas empregou-se caldo seletivo enriquecido PPLO e BHI, O descongelamento dos isolados foi sucedido de subcultivos em caldos enriquecidos seletivos diferenciais e Agar A7 para caracterização fenotípica. A quantificação, a identificação e o teste de susceptibilidade a antimicrobianos foi realizada através do sistema de triagem Mycofast® Screening Evolution (Elitech Group), cuja identificação se baseia na susceptibilidade bacteriana pelo sistema Identibiotiqué. A susceptibilidade dos isolados foi determinada frente aos antimicrobianos doxiciclina 8 μg/ml, roxitromicina 4 μg/ml e ofloxacina 4 μg/ml, amplamente utilizados no tratamento empírico de pacientes com infecções do trato urogenital. Nosso estudo detectou alto nível de resistência dos isolados armazenados para doxiciclina e ofloxacina. Este é o primeiro estudo envolvendo isolamento e o perfil de susceptibilidade destes agentes realizado na região norte.

The apicomplexan Cryptosporidium is a protozoan parasite of humans and other mammals. Cryptosporidium species cause acute gastro-enteritis and diarrheal disease in healthy humans and animals, and cause life-threatening infection in immuno-compromised individuals such as people with AIDS. It has a one-host life cycle and invades intestinal epithelial cells causing diarrhea, or more rarely the pulmonary epithelium. Cryptosporidium carries out all the asexual reproductive stages like several other apicomplexans. Current annotation of this organism predicts it to contain 3884 genes of which only 1581 genes have predicted functions. By using a combination of bioinformatics analysis, biochemical evidence, and high-throughput data, a genome-scale metabolic model of Cryptosporidium hominis is being constructed. The current model is comprised of approximately 213 gene-associated enzymes involved in major metabolic pathways including carbohydrate, nucleotide, amino acid, and energy metabolism. The approach of constructing a genome-scale model provides a link between the genotype and the phenotypic behavior of the organism, making it possible to study and predict behavior based upon genome content. This modeling approach provides an overview for evaluating missing components in a metabolic network and provides an analytical framework for interpreting data as more research becomes available. The goal of constructing this model is to systematically study and analyze various functional behaviors of C. hominis with respect to its stages in life cycle and pathogenicity.

Mycoplasma hominis est un pathogène humain opportuniste responsable d’infections génitales et néo-natales. Modifier génétiquement cette bactérie est nécessaire afin de comprendre les mécanismes de virulence et d’infection de ce pathogène. Il n’existe à ce jour aucun outil moléculaire efficace permettant de manipuler le génome de M. hominis, limitant les recherches sur sa pathogénicité et son métabolisme particulier reposant sur l’arginine. De nouvelles technologies rassemblées sous le terme de Biologie de Synthèse (BS) ont récemment émergé, offrant des perspectives inédites pour l’étude des mycoplasmes en permettant de modifier leurs génomes à grande échelle et de produire des souches mutantes. Ces travaux menés au J. Craig Venter Institute (JCVI, USA) ont montré que le génome de mycoplasmes apparentés pouvait être cloné et manipulé dans la levure avant d’être transplanté dans une cellule receveuse. La levure sert d’hôte d’accueil temporaire pour modifier le génome de la bactérie. Cette approche novatrice ouvre de nombreuses perspectives dans le cadre du développement de la génomique fonctionnelle chez les mycoplasmes pour lesquels les outils génétiques efficaces sont peu nombreux. Le but de cette thèse a été d’adapter pour la première fois certains outils de BS à M. hominis dans le but de créer des mutants déficients pour une fonction donnée. Pour cela, le génome de la souche type de M. hominis PG21 (665 kb) a été cloné dans la levure Saccharomyces cerevisiae par « Transformation-Associated Recombination cloning » (TAR-cloning). Deux clones (B3-2 et B3-4) de levure possédant le génome complet de M. hominis ont été validés par analyse en PCR simplex, PCR multiplex et électrophorèse en champs pulsé (PFGE). Ces clones levures ont ensuite été propagés en milieu sélectif durant 180 générations (30 passages), afin d’évaluer la stabilité du génome bactérien dans son hôte. Cette expérience a montré que (i) si la taille du génome de M. hominis ne variait pas au cours des premiers passages, elle diminuait progressivement à partir du dixième passage (≈60 générations), et que (ii) les zones du génome enrichies en séquence répétées étaient préférentiellement perdues. En tenant compte de ces résultats, le génome de M. hominis a été modifié chez le clone B3-4 par la technique « Clustered Regularly Interspaced Short Palindromic Repeats/Cas9 » (CRISPR/Cas9) lors de passages précoces. Des clones de S. cerevisiae possédant un génome de M. hominis PG21 complet délété du gène vaa, codant une protéine d’adhésion majeure, ont été ainsi produits. La dernière étape de cette approche consistait à transplanter le génome modifié dans une cellule receveuse de M. hominis ou de Mycoplasma arthritidis, espèce phylogénétiquement la plus proche de M. hominis. Aucun protocole de transformation de M. hominis n’étant disponible au début de nos travaux, cette étape constituait un verrou majeur dans la mise en place des outils de BS chez cette espèce. Ce verrou a été en partie levé puisqu’une méthode de transformation de M. hominis basée sur du polyéthylène glycol (PEG) et mettant en jeu le plasposon pMT85 (plasmide contenant un transposon conférant la résistance à la tétracycline) a été mise au point au laboratoire. Cette technique de transformation, développée pour la souche de référence M. hominis M132 (745 kb) reste encore peu efficace ; elle est néanmoins reproductible et a permis d’obtenir des mutants d’intérêt de M. hominis. Le transformant n°28-2 a, ainsi, été muté dans le gène Mhom132_2390, codant le précurseur de la protéine P75, une adhésine putative de M. hominis. Le séquençage des génomes complets d’autres transformants a révélé l’insertion de multiples copies du transposon et la présence d’évènements de duplication et d’inversion de larges fragments d’ADN dans au moins deux génomes de M. hominis
Mycoplasma hominis is an opportunistic human pathogen responsible for genital and neonatal infections. Genetically modifying this bacterium is necessary to understand the virulence and infection mechanisms of this pathogen. There is currently no effective molecular tool to engineer the genome of this bacterium, limiting research on its pathogenicity and its peculiar metabolism based on arginine.New technologies have recently emerged in the field of Synthetic Biology (BS), offering new perspectives for the study of mycoplasmas by allowing large scale genome modifications and the production of mutant strains. Work at the J. Craig Venter Institute (JCVI, USA) has shown that the genome of related mycoplasmas can be cloned and manipulated in yeast before being transplanted into a recipient cell. The yeast serves as a temporary host to modify the genome of the bacterium. This innovative approach opens many perspectives in the development of functional genomics in mycoplasmas for which there are few effective genetic tools. The goal of this thesis was to adapt a number of BS tools to M. hominis for the first time, in order to create mutants deficient for a given function. To achieve this goal, the genome of the M. hominis type strain PG21 (665 kb) was cloned into the yeast Saccharomyces cerevisiae by Transformation-Associated Recombination cloning (TAR-cloning). Two yeast clones (B3-2 and B3-4) possessing the complete genome of M. hominis were validated by simplex PCR, multiplex PCR and Pulsed Field Gel Electrophoresis (PFGE) analyses. These yeast clones were then propagated in a selective medium for 180 generations (30 passages) to evaluate the stability of the bacterial genome in its host. This experiment showed that (i) the size of the genome of M. hominis did not change during the first passages, it decreased progressively from the tenth passage (≈60 generations), and (ii) the enriched genome areas in repeated sequence were preferentially lost. Thus, the genome of M. hominis was modified in the B3-4 clone at early passages using the Clustered Regularly Interspaced Short Palindromic Repeats/Cas9 (CRISPR/Cas9) technology. Yeast clones with a complete M. hominis PG21 genome with a deleted vaa gene, encoding a major adhesion protein, were produced using this approach. The final step of this approach was to transplant the modified genome into a recipient cell of M. hominis or Mycoplasma arthritidis, the species phylogenetically closest to M. hominis. As no M. hominis transformation protocol was available at the beginning of our work, this step constituted a major obstacle in the implementation of BS tools in this species. This barrier has been partially lifted since a method of transformation of M. hominis based on polyethylene glycol (PEG) and involving the plasposon pMT85 (plasmid carrying a transposon conferring resistance to tetracycline) has been developed in the laboratory. This transformation technique, developed for the reference strain M. hominis M132 (745 kb) still remains not very efficient; it is nevertheless reproducible and allowed to obtain M. hominis mutants of interest. The Mhom132_2390 gene, encoding the precursor of the P75 protein, a putative adhesin of M. hominis, was effectively mutated in transformant No. 28-2. Complete genome sequencing of other transformants revealed the insertion of multiple copies of the transposon and the presence of duplication and inversion of large DNA fragments within at least two M. hominis genomes.In conclusion, this data has opened the way for the development and transposition of existing genetic modification approaches to M. hominis, previously considered as a genetically intractable bacterium

Mycoplasmas represent some of the smallest and simplest free-living organisms known. Mycoplasma hominis and Mycoplasma pneumoniae are two human pathogens that colonise the urogenital and respiratory tracts, respectively, causing a diverse range of disease. Detection of Mycoplasma hominis is hampered by the fastidious nature and genetic heterogeneity of this organism. Characterisation of mycoplasmas is becoming more important due to increasing antibiotic resistance, particularly in M. pneumoniae, and the need for more discriminatory methods to enhance epidemiology and examine transmission chains. Firstly, this thesis develops a quantitative, multiplex, real-time PCR assay to simultaneously detect M. hominis and Ureaplasma species in neonatal clinical specimens, where infection is associated with chronic lung disease, bacteremia and other clinical signs. Results showed that the PCR method was clinically more sensitive than culture and has applications for monitoring bacterial load in clinical specimens and characterising bacterial response to antibiotics. Secondly, genetic characterisation of M. hominis was undertaken by the examination of the variable adherence-associated antigen and the development of sequence based typing. Due to the genetic heterogeneity of M. hominis, bioinformatics analysis of genomic sequence was used as a novel method to develop a minimum multi-locus sequence typing (MLST) scheme that accurately represented genomic phylogeny of this species. Finally, an MLST scheme was developed for M. pneumoniae, to aid the analysis of epidemic periods and clusters of infection. A successful scheme was developed based on eight housekeeping genes which had increased discrimination of M. pneumoniae compared to established typing methods for this organism. Furthermore, the MLST scheme was found to be representative of genomic sequence-derived phylogeny, with two distinct genetic clades identified. Application of this MLST to UK epidemics revealed that no predominant sequence type was responsible for the epidemic periods studied, indicating a polyclonal population, supporting the hypothesis that epidemics are driven by population immunity.

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Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico
We performed a combination of proteinase assay, either in solution or immobilized in sodium dodecyl sulfate-polyacrylamide gel copolymerized with gelatin, to detect and quantify proteinases of Dermatobia hominis second (L2) and third (L3) instar larvae. In quantitative assays, we examined proteinase activity by hydrolysis of a panel of peptide bonds specific for the main proteinase classes. We verified that the pGlu-Phe-Leu p-nitroanilide substrate was hydrolyzed by crude extracts of L2 (3.0 ? 0.2 nmoles hour-1 mg of protein-1) and L3 (7.7 ? 0.1 nmoles hour-1 mg of protein-1) and that both activities were partially inhibited by transepoxysuccinyl- L-leucylamido-(4-guanidino)butane, 15 % and 3 % respectively. Also, we demonstrated that the Na-p-Tosyl-L-Arg methyl ester substrate was hydrolyzed by crude extracts of L2 (117 ? 24 nmoles hour-1 mg of protein-1) and L3 (111 ? 10 nmoles hour-1 mg of protein-1), suggesting a predominance of esterase activity in the crude larval preparation. Interestingly, the specific activity of serine-proteinases was totally inhibited by Phenylmethylsulphonyl fluoride in the L3 crude extract, while only 10 % of this enzyme class activity was inhibited in the L2 crude extract. Also, we have detected crude extract L2 (Km = 7,59) larvae have more affinity than L3 larvae (Km = 35,75) to Na-p-Tosyl-L-Arg methyl ester. The results of the qualitative assays with substrate gels suggested that L2 and L3 larvae express serine-proteinases with similar (13 kDa and 22 kDa) and distinct (50 kDa in L2 and 30 kDa in L3) relative molecular masses. Additionally, we have isolated an enriched esterase activity from L3 crude extract using successive chromatographies in Aprotinine-Agarose and DEAE-Sephacell columns. By this strategy we detected only one 50 kDa proteinase in this larvae crude extract. Finally, these findings contribute to the biochemical characterization of D. hominis L2 and L3 larvae.
Neste trabalho foram realizados ensaios de atividade de proteinase em solu??o e com prote?nas imobilizadas em gel de poliacrilamida contendo dodecil sulfato de s?dio copolimerizado com gelatina, para detec??o e quantifica??o das proteinases presentes nos extratos larvares de segundo (L2) e terceiro (L3) est?gios de Dermatobia hominis. Nos ensaios quantitativos, utilizou-se um painel de pept?deos sint?ticos espec?ficos para as principais classes de proteinases. Verificamos que o substrato pGlu-Phe-Leu p-nitroanilide foi hidrolisado pelo extrato total de L2 (3,0 ? 0,2 nmoles hora-1 mg de prote?na-1) e L3 (7,7 ? 0,1 nmoles hora-1 mg de prote?na-1) e que ambas atividades foram parcialmente inibidas pelo trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane, 15 % e 3 % respectivamente. Tamb?m, demonstramos que o substrato Na-p-Tosyl-L-Arg methyl ester foi hidrolisado pelos extratos totais de L2 (117 ? 24 nmoles hora-1 mg de prote?na-1) e L3 (111 ? 10 nmoles hora-1 mg de proteina-1), sugerindo uma predomin?ncia da atividade ester?sica nestes extratos. A atividade espec?fica de serino-proteinases foi totalmente inibida pelo phenylmethylsulphonyl fluoride nos extratos de L3, enquanto que somente 10 % desta atividade foi inibida nos extrados de L2. Al?m disso, n?s detectamos que o extrato total das larvas L2 (Km = 7,59) tem maior afinidade ao Na-p-Tosyl-L-Arg methyl ester do que o extrato total das larva de L3 (Km = 35,75). Os resultados do ensaio qualitativo com g?is de substrato sugerem que os extratos larvares L2 e L3 expressam serino-proteinases com similares (13 kDa e 22 kDa) e distintas (50 kDa em L2 e 30 kDa em L3) massas moleculares relativas. Adicionalmente, isolamos uma atividade ester?sica enriquecida do extrato total de L3 utilizando sucessivas cromatografias em colunas de Aprotinina-Agarose e DEAE-Sephacell. Com esta estrat?gia, detectamos somente uma banda de proteinase de 50 kDa neste extrato total. Estes resultados contribuem para a caracteriza??o das proteinases larvares de D. hominis.

Ce travail essaie de montrer comment la notion de dignite de l'homme s'inscrit dans les essais. Dans un premier temps, nous avons mis en evidence, par une analyse diachronique et structurale, les presupposes ideologiques regissant cette notion, de maniere a en percevoir les traces dans les discours de la renaissance. La dignitas hominis apparait comme un enonce capable d'etre utilise dans tous les genres discursifs. Une fois ces premisses posees, nous avons montre comment cet enonce etait, tout d'abord, deconstruit dans les essais. Pour ce faire, nous avons analyse le travail de "grammairien" de montaigne en relevant les occurrences et l'utilisation des motifs constitutifs de l'enonce de la dignitas hominis. Tout le travail critique de montaigne tend a reinscrire l'homme dans une limite pour pouvoir redefinir une autre grandeur humaine. Ainsi, dans un troisieme temps, nous avons mis en lumiere la reconstruction de la valeur de l'excellence de l'homme chez montaigne; nouvelle valeur qui temoigne d'une double modification dans l'histoire des mentalites. La dignite de l'homme est liee a la prise en compte de l'alterite et elle ne trouve son fondement que dans la recherche du convenable personnel. L'excellence de l'homme n'est donc plus, apres montaigne, un enonce heteronome mais participe de la construction de soi
This study aims at showing how montaigne deals with the concept of man's dignity in his essais. A diachronic and structural analysis outlines and traces in the writings of the renaissance the ideological presuppositions of dignity at work prior to montaigne. Once having established the idea that dignitas hominis is an utterance potentially present in every type of speech, the utterance and its constituant motifs are shown to undergo deconstruction in the essais through montaigne's " grammarian " processing. His critical work aims at rewriting man's place within new boundaries in order to redefine human grandeur. Therefore, montaingne's reconstruction of man's excellence is brought to light as a new value testifying to a double change in the history of ideas. Human dignity is a concept connected with an awareness of other, and it finds its roots solely in the quest for personal appropriateness. Man's excellence after montaigne can no longer be construed as a heteronomous utterance but as part of the building of self

Orientador : Ana Maria Lima de Azeredo-Espin
Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia
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Resuimo: Dermatobia hominis (Linnaus Jr.) conhecida popularmente como mosca do berne. É uma das principais causadoras de miíase primária em vertebrados. sendo uma das principais pragas da pecuária nacional. causando grandes prejuízos à industria coureira. laticínios e frigoríficos. A mosca do berne apresenta uma biologia interessante e ciclo de vida bastante peculiar. o que dificuta a observação da espécie na natureza e a manutenção desta em laboratório. Por estes motivos. a caracterização da variabilidade genética e da estrutura populacional são importantes ferramentas na compreensão da evolução de D. hominis. Para este fim. foram utilizadas as técnicas de RAPD, com seis populações, e de RFLP do DNA mitocondrial, com doze populações. Os resultados foram comparados entre si e com estudos anteriores. Além disso, foram comparadas diferentes abordagens que podem ser aplicadas aos dados obtidos com RAPD. Os resultados de RAPD e RFLP do DNA mitocondrial sugerem que a espécie comporta-se como uma metapopulação. na qual as populações são altamente variáveis e apresentam fluxo gênico entre si. Quanto às diferentes abordagens aplicáveis aos dados de RAPD foi observado que as medidas de Dice e Jaccard, calculadas a partir da matriz completa. e a distância de Nei (1972) calculada com as modificações de Lynch & Milligan (1994), foram as mais adequadas ao estudo da variabilidade de D. hominis. Com o uso do RFLP do mtDNA foram detectados setenta e dois baplótipos, o que denota alta variabilidade para a espécie
Abstract: Dermatobia hominis (Linnaus Jr.), the human bot fly, is one of the most important agent of primary myiases in vertebrates and one of the most important pests of the national cattle breeding and causes large losses in the industry of leather, milk and meat. The bot fly has an interesting biology and a very peculiar life cycle. These facts make difficult to observe the specie in nature and to breed it in laboratory. Due to these reasons, the characterization of the genetic variability and structure are important tools in the understanding of the D. hominis evolution. Techniques based on RAPD, with six populations, and RFLP of mitochondrial DNA, with twelve populations were applied for this purpose. Estimates of several biometrical procedures applied to the data were compared. Comparisons were also made with results of previous studies. Different approaches for analyzing RAPD data were also compared. The results of RAPD and mtDNA suggest that the specie behave as a metapopulation whose populations are variable and present gene flow among themselves. Results of different methods based on RAPD indicated that the Dice and Jaccard measures, calculated :from the complete matrix of data, and of Nei distances (1972) with the modifications of Lynch & Milligan (1994) were more adequate for the study of the variability of D. hominis. With the use of rntDNA RFLP, seventy-two haplotypes were detected, indicating high variability for this specie
Mestrado
Genetica Animal e Evolução
Mestre em Genética e Biologia Molecular

Orientador: Teresa Cristina Goulart de Oliveira Sequeira
Resumo: O presente trabalho foi desenvolvido com o objetivo de caracterizar os produtos de excreção/secreção (PE/S) de larvas de Dermatobia hominis, especialmente, no que se refere à atividade proteolítica desses produtos. Os PE/S foram obtidos de larvas de primeiro estágio (L1) cultivadas em laboratório e de larvas de segundo (L2) e terceiro (L3) estágios colhidas de bovinos parasitados. O perfil das proteínas foi obtido pela eletroforese dos PE/S em gel de poliacrilamida contendo sódio dodecil sulfato (SDS-PAGE) e a atividade proteolítica foi investigada utilizando gelatina, azocaseína e N-a-benzoil-arginina-nitroanilida (BAPNA) como substratos. Para caracterização das proteases foram realizados ensaios utilizando estes mesmos substratos, nos quais as amostras foram tratadas com os inibidores: PMSF, TPCK, TLCK, DCI, E-64, EDTA, Elastatinal, Leupeptina, Fenantrolina e Antipaína. Nos PE/S de L1 foram detectadas exclusivamente proteases com peso molecular aparente acima de 168 kDa, cuja inibição revelou pertencerem aos grupos das metalo-proteases e serina-proteases. Os zimogramas dos PE/S de L2 e L3 em géis copolimerizados com gelatina revelaram uma ampla faixa de atividade proteolítica, na qual estavam presentes proteases de alto, médio e baixo pesos moleculares aparentes. Nos ensaios realizados com inibidores de proteases, utilizando os três substratos, as proteases presentes em L2 e L3 foram identificadas como pertencentes ao grupo das serina-proteases, sendo em L3, predominantemente, do tipo tripsina. As alterações nos padrões de proteólise e nas características bioquímicas das proteases presentes nos três estágios larvais discuti- se à atividade das larvas em cada fase do seu desenvolvimento.
Abstract: In the present investigation the biochemical characteristics of the Dermatobia hominis larvae secretory/excretory products (PE/S) were analyzed mainly regard to their proteolytic activities. The PE/S of first-instar larvae were collected from larvae reared in the laboratory and of the second and third instars from larvae removed from infested cattle.The PE/S were tested in a sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) to identify their proteic profiles and the proteases activity were investigated using gelatine, azocasein, and N-a-benzoil-arginine-nitroanilide (BAPNA) as substrates. The proteases characterizations were performed by treating PE/S samples by the following proteases inhibitors: PMSF, TPCK, DCI, E-64, EDTA, Elastatinal, Leupeptine, Phenatroline, and Antipain. SDS-PAGE-Gelatin profiles of L1 PE/S revealed only proteases with molecular weight above 168 KDa whereas profiles from L2 and L3 PE/S revealed a high range of proteolytic activity, including high, medium and low molecular weight proteases. The assays performed with the protease inhibitors, and gelatin and azocasein as substrates, revealed that metalloprotease is the major class of proteases present in the PE/S of L1 besides the low content of serine-proteases. The major class of proteases present in the PE/S of L2 and L3 was characterized as serine-proteases, being in L3 predominantly of trypsin type. The changes in the proteolytic patterns and biochemical characteristics of the proteases found in all three instar larvae of D. hominis were with the larval activity in each phase of their development.
Mestre

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The aim of this study was to characterize the association between the seasonality of dermatobiosis in bovines and its dipterans potential vectors as well as evaluate the association among the counts of Dermatobia hominis larvae and of potential vectors with meteorologicals data. It also had the objective of delimiting, quantifying and characterizing the geographical space of Serop?dica and Itagua? municipalities in relation to the favorable degrees for the occurrence of dermatobiosis in bovines in the rainy and dry periods using the Vista SAGA? 2007 information system. The seasonal fluctuation of D. hominis larvae was obtained by counts in ten bovines and of muscoids dipterans by capture using Adultrap? traps with sardine bait, once a month each, in three properties named A, B and C, localized in the Serop?dica, Paracambi and Itagua? municipalities, respectively, in the period of January 2006 to December 2007. The meteorological data of compensated mean temperature, pluvial precipitation and relative humidity were obtained in the Agricultural Ecology Station (22?48'S, 43?41'W and 33m of altitude). The associations between D. hominis larvae fluctuations, adult dipterans and climatological data were verified using the Spearman correlation test. Kruskal-Wallis and Mann-Whitney tests were used in the analysis of the significant differences between the rainy and dry periods in each property in the monthly average of D. hominis larvae, in the total monthly number and of dipterans family and in the analysis of other studies. In the elaboration of models by geoprocessing, some abiotic factors were used: occupation and covering of the land, altitude, declivity, soils and climatic factors, in which the evaluation, signature and monitorship functions of the Vista SAGA? 2007 program were applied. The infestation by D. hominis larvae in bovines was present during the 24 months of study in the three properties. A total of 18.966 dipterans potential vectors was captured, being 30,16% in the A property, 34,59% in the B property and 35,25% in the C property but there were no significant statistic difference between these. There was a positive and significant correlation (rs=0,63) between monthly averages of D. hominis larvae and relative humidity in 2006 at the A property; there was a positive and significant correlation between the total number of captured dipterans and pluvial precipitation (rs=0,80) and temperature (rs=0,60) at the B and C properties, respectively, in 2007. It was also observed a positive and significant correlation between the total number of captured dipterans of the Sarcophagidae family (rs=0,60) with the temperature in the C property in 2006, and between Muscidae (rs=0,67) and Calliphoridae (rs=0,76) with pluvial precipitation in the B property in 2007. There wasn t an association between monthly averages of D. hominis larvae and dipterans. It was verified a pattern lack in the association between monthly and periods fluctuations, rainy and dry of D. hominis larvae with the dipterans potential vectors fluctuations and of both with meteorologicals data. The geoprocessing allowed delimiting, quantifying and characterizing the potential of space temporal distribution of dermatobiosis in bovines.
Este estudo teve como objetivos caracterizar a associa??o entre a sazonalidade da dermatobiose em bovinos e seus d?pteros potenciais vetores, assim como avaliar a associa??o entre as contagens de larvas de Dermatobia hominis e de potenciais vetores com os dados meteorol?gicos. Teve tamb?m como objetivo, delimitar, quantificar e caracterizar o espa?o geogr?fico dos munic?pios de Serop?dica e Itagua? quanto aos gradientes de favorabilidade para a ocorr?ncia da dermatobiose em bovinos nos per?odos chuvoso e seco utilizando o sistema de informa??o Vista SAGA? 2007. A flutua??o sazonal de larvas de D. hominis foi obtida por contagens em dez bovinos e a de d?pteros musc?ides por captura, usando armadilhas Adultrap? com isca de sardinha, ambos uma vez ao m?s, nas tr?s propriedades denominadas A, B e C, localizadas nos munic?pios de Serop?dica, Paracambi e Itagua?, respectivamente, no per?odo de janeiro de 2006 a dezembro de 2007. Os dados meteorol?gicos de temperatura m?dia compensada, precipita??o pluvial e umidade relativa, foram obtidos na Esta??o Ecologia Agr?cola (22?48'S, 43?41'W e 33m de altitude). As associa??es entre as flutua??es de larvas de D. hominis, d?pteros adultos e dados climatol?gicos foram verificadas utilizando-se o teste de correla??o de Spearman. Os testes de Kruskal-Wallis e Mann-Whitney foram empregados na an?lise das diferen?as significativas entre os per?odos, chuvoso e seco, por propriedade, para a m?dia mensal de larvas de D. hominis, o n?mero mensal total e por fam?lia de d?pteros e na an?lise de dados publicados em outros estudos. Para a elabora??o dos modelos por geoprocessamento, utilizaram-se os fatores abi?ticos: uso e cobertura das terras, altitude, declividade, solos e fatores clim?ticos, aplicando-se as fun??es avalia??o, assinatura e monitoria do programa Vista SAGA? 2007. A infesta??o por larvas de D. hominis em bovinos esteve presente ao longo dos 24 meses de estudo nas tr?s propriedades. Um total de 18.966 d?pteros potenciais vetores foi capturado. Deste, 30,16% foram capturados na propriedade A, 34,59% na propriedade B e 35,25% na propriedade C, mas sem diferen?a estat?stica significativa entre estas. Houve correla??o positiva e significativa (rs=0,63) entre as m?dias mensais de larvas de D. hominis e umidade relativa, no ano de 2006, na propriedade A; houve correla??o positiva e significativa entre o n?mero total de d?pteros capturados e precipita??o pluvial (rs=0,80) e tamb?m com a temperatura (rs=0,60) nas propriedades B e C, respectivamente, no ano de 2007. Observou-se tamb?m correla??o positiva e significativa entre o n?mero total de d?pteros capturados da fam?lia Sarcophagidae (rs=0,60) com a temperatura na propriedade C no ano de 2006 e entre Muscidae (rs=0,67) e Calliphoridae (rs=0,76) com precipita??o pluvial na propriedade B em 2007. N?o ocorreu associa??o entre as m?dias mensais de larvas de D. hominis e de d?pteros. Constatou-se uma falta de padr?o na associa??o entre as flutua??es mensais e por per?odos, chuvoso e seco, das larvas de D. hominis com as flutua??es de d?pteros potenciais vetores e de ambas com os dados meteorol?gicos. O geoprocessamento permitiu delimitar, quantificar e caracterizar o potencial de distribui??o espa?o-temporal da dermatobiose nos per?odos chuvoso e seco.

Esta tesis plantea una doble propuesta: creativa y teórica. Basada en la obra artística personal del autor (un descarnado diario personal autobiográfico realizado durante más de una década), se realiza un profundo estudio teórico de la citada obra desde un punto de vista fundamentalmente semiótico consiguiendo no sólo argumentar específicamente la citada obra sino también profundizar en el estudio general de los conceptos indicialidad, espacio y tiempo, fundamentales en cualquier concepción rigurosa de la fotografía y ejes nucleares del proyecto artístico que vertebra esta tesis.

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Universidade Estadual Paulista (UNESP)
O presente trabalho foi desenvolvido com o objetivo de caracterizar os produtos de excreção/secreção (PE/S) de larvas de Dermatobia hominis, especialmente, no que se refere à atividade proteolítica desses produtos. Os PE/S foram obtidos de larvas de primeiro estágio (L1) cultivadas em laboratório e de larvas de segundo (L2) e terceiro (L3) estágios colhidas de bovinos parasitados. O perfil das proteínas foi obtido pela eletroforese dos PE/S em gel de poliacrilamida contendo sódio dodecil sulfato (SDS-PAGE) e a atividade proteolítica foi investigada utilizando gelatina, azocaseína e N-a-benzoil-arginina-nitroanilida (BAPNA) como substratos. Para caracterização das proteases foram realizados ensaios utilizando estes mesmos substratos, nos quais as amostras foram tratadas com os inibidores: PMSF, TPCK, TLCK, DCI, E-64, EDTA, Elastatinal, Leupeptina, Fenantrolina e Antipaína. Nos PE/S de L1 foram detectadas exclusivamente proteases com peso molecular aparente acima de 168 kDa, cuja inibição revelou pertencerem aos grupos das metalo-proteases e serina-proteases. Os zimogramas dos PE/S de L2 e L3 em géis copolimerizados com gelatina revelaram uma ampla faixa de atividade proteolítica, na qual estavam presentes proteases de alto, médio e baixo pesos moleculares aparentes. Nos ensaios realizados com inibidores de proteases, utilizando os três substratos, as proteases presentes em L2 e L3 foram identificadas como pertencentes ao grupo das serina-proteases, sendo em L3, predominantemente, do tipo tripsina. As alterações nos padrões de proteólise e nas características bioquímicas das proteases presentes nos três estágios larvais discuti- se à atividade das larvas em cada fase do seu desenvolvimento.
In the present investigation the biochemical characteristics of the Dermatobia hominis larvae secretory/excretory products (PE/S) were analyzed mainly regard to their proteolytic activities. The PE/S of first-instar larvae were collected from larvae reared in the laboratory and of the second and third instars from larvae removed from infested cattle.The PE/S were tested in a sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) to identify their proteic profiles and the proteases activity were investigated using gelatine, azocasein, and N-a-benzoil-arginine-nitroanilide (BAPNA) as substrates. The proteases characterizations were performed by treating PE/S samples by the following proteases inhibitors: PMSF, TPCK, DCI, E-64, EDTA, Elastatinal, Leupeptine, Phenatroline, and Antipain. SDS-PAGE-Gelatin profiles of L1 PE/S revealed only proteases with molecular weight above 168 KDa whereas profiles from L2 and L3 PE/S revealed a high range of proteolytic activity, including high, medium and low molecular weight proteases. The assays performed with the protease inhibitors, and gelatin and azocasein as substrates, revealed that metalloprotease is the major class of proteases present in the PE/S of L1 besides the low content of serine-proteases. The major class of proteases present in the PE/S of L2 and L3 was characterized as serine-proteases, being in L3 predominantly of trypsin type. The changes in the proteolytic patterns and biochemical characteristics of the proteases found in all three instar larvae of D. hominis were with the larval activity in each phase of their development.

Objetivo: Determinar la prevalencia de infección por Strongyloides stercolaris y otras enteroparasitosis en el pueblo joven Santo Toribio de Mogrovejo de Chiclayo durante Junio-Octubre del 2011. Material y Métodos: Estudio descriptivo, trasversal; muestreo aleatorio, estratificado, polietápico, siendo el tamaño muestral de 106 pobladores. Se diseñó y validó una ficha de recolección epidemiológica. Un biólogo recibió entrenamiento en las técnicas de diagnóstico de Strongyloides stercolaris en un centro referencial de Lima. Se recolectaron 3 muestras por paciente, sometidas a 5 técnicas parasitológicas: Examen directo de heces, Baermann modificado en Copa por Lumbreras, Test de sedimentación espontánea, Cultivo en agar y Cultivo Dancescu. Resultados: Se visitaron 124 casas; el porcentaje de respuesta fue de 85,7%. Se logró entrevistar 106 personas. El promedio de edad fue de 27,8 +/- 16,9 años; hubieron 31 hombres (29,2%) y 75 mujeres (70,8%). El 26,4% de personas habían realizado un viaje a la Sierra y/o Selva en los últimos 5 años con una estancia mayor a un mes. El piso de tierra fue el más frecuente en el total de viviendas (55,6%); 102 personas (96,2 %) tenían desagüe; 23 pobladores (21,7 %) tuvieron al menos un parásito detectado. No se hallaron pobladores infectados con Strongyloides stercolaris. La enteroparasitosis más frecuente fue por protozoarios, con predominio de Blastocystis hominis en un 12,3%. Conclusiones: Se halló una baja frecuencia de enteroparasitosis y ausencia de pobladores infectados con Strongyloides stercolaris. El parásito más frecuente fue Blastocystis hominis.
Tesis

O presente estudo teve como objetivo a caracterização genômica das espécies: Mycoplasma hominis, Ureaplasma urealyticum e Ureaplasma parvum em amostras cervicais de gestantes. Cepas de Mycoplasma hominis e de Ureaplasma spp., foram identificadas através do cultivo e por PCR utilizando-se primers genéricos e específicos. Os achados na literatura são contraditórios quanto a participação dessas bactérias nas doenças humanas, especialmente quanto a participação destas em casos de infertilidade e alterações perigestacionais. Considerando a marcante heterogeneidade das cepas de Mycoplasma hominis, utilizamos a reação de RAPD com a finalidade de definir os perfis genômicos das cepas isoladas. A caracterização dos biótipos 1 e 2 do gênero Ureaplasma, respectivamente, Ureaplasma urealyticum e Ureaplasma parvum, através da análise da MBA (Múltipla Banda Antigênica) por PCR, teve como finalidade determinar a predominância de cada uma dessas espécies. Foram estudadas amostras de material cervical de 163 gestantes em várias idades gestacionais. Os resultados obtidos neste trabalho foram: M. hominis foi detectado em 14,7% (24/163) das amostras, sendo a RAPO realizada em 7 das amostras positivas e puras de M. hominis. A reação RAPD demonstrou similaridade do perfil gênico entre as amostras 3 e 4 e entre as amostras 6 e 8, enquanto que as amostras 2, 5 e 7 demonstraram uma grande heterogeneidade. O gênero Ureaplasma foi isolado em 54,6% (89/163) das amostras, sendo 31,5% (28/89) cepas de U. urealyticum e 68,5% (61/89) U. parvum. Foram detectados 10,4% (17/163) casos de co-infecções M. hominis e Ureaplasma spp. O estudo da suscetibilidade das cepas isoladas frente a tetraciclina foi realizado com o intuito de se determinar a freqüência de cepas resistentes a essa droga. Considerando ser este antibiótico o de eleição no tratamento das micoplasmoses humanas e devido aos altos índices de resistência apresentados, resistência esta conferida pela presença do plasmídeo tetM, submetemos nossas amostras à pesquisa deste plasmídeo através da PCR. A presença do plasmídeo tetM foi detectada em 29,5% (7/24) das amostras positivas para Mycoplasma hominis e em 60,7% (54/89) das amostras positivas para Ureaplasma spp., sendo que dentre estas, 57,4% (31/54) foram detectados na espécie U. parvum e 42,6% (23/54) na espécie Ureaplasma urealyticum. O estudo da heterogeneidade intra-espécie das cepas de Mycoplasma hominis pela técnica de RAPD é inédito no Brasil. Da mesma maneira a caracterização das espécies do gênero Ureaplasma em U. parvum e U. urealyticum, através da PCR pela análise da MBA, foi pioneiramente realizada pelo nosso grupo de pesquisa. Esperamos com esse trabalho poder contribuir para um conhecimento melhor da freqüência dessas espécies na nossa população e tentar explicar as divergências das avaliações nas interações entre os micoplasmas e o hospedeiro.
The objective of this study was to characterize the species Mycoplasma hominis, Ureaplasma urealyticum and Ureaplasma parvum in cervical samples from pregnant women. Strains of Mycoplasma hominis and Ureaplasma spp., were identified by means of culture and by PCR using generic and specific primers. The findings in the literature with regard to the involvement of these bacteria in human diseases are contradictory, especially regarding their involvement in cases of infertility and perigestational changes. In view of the strong heterogeneity among Mycoplasma hominis strains, we used RAPD to define the profiles of the strains isolated. Biotypes 1 and 2 of the genus Ureaplasma - Ureaplasma urealyticum and Ureaplasma parvum respectively - were characterized by means of MBA (Multiple Banded Antigenic) analysis using PCR to determine the predominance of each of these species. Samples of cervical material from 163 pregnant women at various stages of pregnancy were studied. The results obtained in this study were: M. hominis was detected in 14.7% (24/163) of the samples, and RAPD analyses carried out in 7 M. hominis strains. The RAPD similarly gene profile was showed: between strains 3 and 4, and between strains 6 and 8. The strains 2, 5 and 7 showed a pronounced heterogeneity in the gene profile. The genus Ureaplasma was isolated in 54.6% (89/163) of the samples, of which 31.5% (28/89) were U. urealyticum and 68.5% (61/89) U. parvum. Mycoplasma hominis and Ureaplasma spp., coinfection were detected in 10.4% (17/163) of the samples. The study of the susceptibility of the isolated strains to tetracycline was carried out with a view to determining the frequency of strains resistant to this drug. As this is the antibiotic of choice in the treatment of human mycoplasmoses, and in view of the high resistance indices observed, which are conferred by the presence of the \"tetM\" plasmid, we looked for this plasmid in our samples using PCR. The presence of the tetM plasmid, was detected 57,1% (4/7) of the Mycoplasma hominis - positive samples and in 40,2% (29/72) of the Ureaplasma spp - positive samples. Of the latter, 41,6% (25/72) were detected in the species U. parvum and 33,4% (4/12) were detected in the species Ureaplasma urealyticum. This is the first time that a study of the intraspecies heterogeneity of Mycoplasma hominis strains using RAPD has been carried out in Brazil. Characterization of the genus Ureaplasma species into U. parvum and U. urealyticum by means of PCR using MBA analysis was likewise pioneering effort by our research group. We hope that our work will contribute to a greater knowledge of the frequency of these species in our population and that it will help to explain differences in the assessment of the interaction between mycoplasma and the host.

Blastocystis is an important parasite that infects humans and a wide range of animals like rats, birds, reptiles, etc. infecting a sum of 60% of world population. It belongs to the Stramenopiles, a Heterologous group that includes for example the Phythophthora infestans the responsible for the Irish potato famine. Previous work had reported the presence of an unusual fusion protein that is composed of two of the main glycolytic enzymes; Triosephosphate isomerase-glyceraldehyde-3-phosphate dehydrogenase (TPI-GAPDH). Little is known about this protein. Blastocystis TPI-GAPDH and Blastocystis enolase were both characterized biochemically and biophysically in this project. The phylogenetic relationships of those two proteins among other members of either Stramenopiles, or other members of the kingdom of life were examined and found to be grouping within the chromalveolates. Our studies revealed that those two proteins, Blastocystis enolase and Blastocystis TPI-GAPDH, had a peptide signal targeting them to the mitochondria. This was an unusual finding knowing that text books always referred to the glycolytic pathway as a canonical cytoplasmic pathway. Structural studies had also been conducted to unravel the unknown structure of the fusion protein Blastocystis TPI-GAPDH. X-ray crystallography had been conducted to solve the protein structure and the protein was found to be a tetrameric protein composed of a central tetrameric GAPDH protein flanked with two dimmers of TPI protein. Solving its structure would be the starting point towards reviling the role that TPI-GAPDH might play in Blastocystis and other organisms that it was found in as well. Although a fusion protein, the individual components of the fusion were found to contain all features deemed essential for function for TPI and GAPDH and contain all expected protein motifs for these enzymes.

La présente thèse consiste dans l'édition princeps, avec introduction, traduction et commentaire, du Liber de miseria hominis de Hugues de Miramar, chartreux à Montrieux au milieu du XIIIe siècle. L'ouvrage est un traité de spiritualité centré sur les thèmes du mépris du monde et de l'excellence de la voie cartusienne, et transmis par deux manuscrits, qui véhiculent deux versions de la même oeuvre, une courte et une longue. L'introduction et le commentaire se sont notamment attachés à exposer les questions critiques posées par les manuscrits, à présenter l'auteur, sa personnalité spirituelle et son oeuvre, et à justifier notre choix de n'éditer ici que la version courte du Liber. Plusieurs points d'intérêt ont été relevés, comme la personnification de la mort, qui tourne l'oeuvre vers la fin du Moyen Âge, ou la teneur autobiographique de l'ouvrage, qui mêle les genres littéraires d'un traité et d'un récit. Le texte est un précieux témoin de la sensibilité religieuse du XIIIe siècle
This thesis is the first edition, with introduction, translation and commentary, of the Liber de miseria hominis by Hugues de Miramar, a Carthusian monk who lived in Montrieux in the middle of the XIIIth century. The work is a spiritual treatise mainly dealing with the themes of world contempt and of the excellence of the carthusian creed, taking the shape of two manuscripts, that offer two versions of the same work, a shorter one and a longer one. In the introduction and the commentary, particular attention was paid to setting out the critical questions raised by the manuscritps, to presenting the author, his spiritual individuality and his work, and to justifying our choice of editing here only the shorter version of the Liber. Several points seemed worthy of interest, namely the personification of death, that sets the work circa the end of the Middle Ages, as well as the autobiographical content of the book, which blends the literary styles of a treatise and a retrospective first-person narrative. The text provides priceless evidence of religious sensitivity in the XIIIth century

A endometriose é uma doença caracterizada pela presença de endométrio fora do útero. O estudo objetivou detectar Mollicutes (M. genitalium, M. hominis, U. urealyticum e U. parvum), HPV e N. gonorrhoeae em amostras de swab endocervical, fluido peritoneal e tecido de biópsia de mulheres com (grupo caso) e sem endometriose (grupo controle) e avaliar os achados com a endometriose. No swab endocervical, prevalências de M. hominis (Mh), M. genitalium (Mg), U. urealyticum (Uu) e HPV foram maiores no grupo caso (43,7%, 14,1%, 8,5% e 5,9% respectivamente) que no grupo controle. No fluido peritoneal também foi maior no grupo caso (Mh: 27,8%; Mg: 40,7%; Uu: 3,7% e HPV: 9,6%) do que o grupo controle. No tecido de biópsia, Mh (5,9%) e Mg (13,2%) foram maiores no grupo caso. M. genitalium no fluido peritoneal foi associado à maior produção de IFN-γ e IL-1β (p < 0,05). O perfil de downregulation de genes da inflamação foi acentuado na presença de M. genitalium. Upregulation ocorreu na presença de M. hominis. Mollicutes podem influenciar na resposta imune na endometriose.
Endometriosis is a disease characterized by the presence of endometrium outside of uterus. This study aimed to detect Mollicutes (M. genitalium, M. hominis, U. urealyticum and U. parvum), HPV and N. gonorrhoeae in samples of endocervical swab, peritoneal fluid and biopsied tissue from women with (case group) and without endometriosis (control group) and evaluate the finds with endometriosis. In swab samples the prevalence of M. hominis (Mh), M. genitalium (Mg), U. urealyticum (Uu) and HPV were higher in case group (43.7%, 14.1%, 8.5% e 5.9% respectively) than the control group. In the peritoneal fluid it was higher in the case group as well (Mh: 27.8%; Mg: 40.7%; Uu: 3.7% e HPV: 9.6%).In the biopsied tissue, Mh (5.9%) and Mg (13.2%) were higher in the case group. M. genitalium in the peritoneal fluid was associated to a higher production of IFN-γ and IL-1β. Downregulation of inflammatory genes were accentuated when M. genitalium was detected. Upregulation occurred when M. hominis was detected. Mollicutes could influence in the immune response on endometriosis.

UVOD: Blastocistis hominis (Bh) je najrasprostranjeniji protist na našoj planeti, ali pri tome najkontraverzniji. Infekcija Bh počinje ingestijom hrane ili tečnosti koja je kontaminirana cističnom formom Bh. Nakon gutanja, iz ciste se razvijaju u debelom crevu čoveka vakuolarne forme protista. Fekalno - oralni prenos je najčešći put širenja infekcije. Oboljenje koje Bh izaziva kod ljudi naziva se blastocistoza. Najčešće inficirani imaju gastrointestinalne tegobe, pre svega bol u trbuhu i proliv. Blastocistoza se danas povezuje sa dva klinička entiteta koji predstavljaju poremećaj rada creva, odnosno sindromom iritabilnog creva i hroničnom inflamatornom bolesti creva (HIBC). CILJ RADA I HIPOTEZE: Predmet istraživanja je da se utvrdi povezanost prisustva infekcije Blastocistis hominisom i postojanja zapaljenja sluzokože debelog creva (kolitisa) kod dece sa gastrointestinalnim tegobama, zatim da se utvrdi udeo dece sa posebnom formom kolitisa, hroničnom inflamatornom bolesti creva, među inficiranim Blastocistis hominisom, a da bi se omogućilo bolje razumevanje blastocistoze kod dece. Osnovne hipoteze u istraživanju su statistički značajno veća učestalost pojave kolitisa i hronične inflamatorne bolesti creva kod dece uzrasta od 1 meseca do 18 godina, hospitalizovane zbog bola u trbuhu i/ili proliva koji su inficirani Blastocistis hominisom, kao i statistički značajno veća učestalost kolitisa u odnosu na hroničnu inflamatornu bolest creva u istom uzorku. MATERIJAL I METODE: Prospektivnim ispitivanjem su obuhvaćeni pedijatrijski bolesnici, hospitalizovani na Odeljenju za gastroenterologiju, hepatologiju i ishranu, Instituta za zdravstvenu zaštitu dece i omladine Vojvodine, zbog bola u trbuhu i/ili proliva, iz čije stolice je dokazan Blastocistis hominis. U toku ispitivanja primenjene su standardne metode uzimanja anamneza od bolesnika, fizički pregledi, odgovarajuće standardne laboratorijske analize krvi i stolice, ultrazvučni pregled abdomena, kolonoskopija i patohistološki pregled biopsija debelog creva. Svi bolesnici su lečeni metronidazolom u trajanju 10 dana, prema važećim terapijskim protokolima. REZULTATI: Ispitivanjem je obuhvaćeno 102 bolesnika, koji su an osnovu patohistološkog nalaza podeljeni u tri grupe: 1. Grupa (bolesnici koji nemaju kolitis, obuhvatila je 4 bolesnika (4.4%)), 2. Grupa – (bolesnici koji imaju nespecifični kolitis, obuhvatila je 56 bolesnika (56.55%)) i 3. Grupa –(bolesnici koji imaju hroničnu inflamatornu bolest, obuhvatila je 42 bolesnika (42.41%)). Među ispitanicima je bio podjednak broj dece muškog i ženskog pola, odnosno 51 dečak i 51 devojčica. Uzrast ispitanika koji imaju infekciju Blastocistisom hominisom se kretao u interval od 11 meseci do 17 godina i 7 meseci. Medijana je iznosila 12.54 godine, a prosečna starost 11.25 godine. Blastocistoza nema sezonski karakter (χ2=0.667; df=3; p=0,881). Značajno više inficiranih Blastocistis hominisom živelo u kući, nego u stanu i posedovalo domaće životinje i/ili kućne ljubimce, ali ne postojanje odgovarajućih higijenskih uslova, kanalizacije i vodovoda nije prediktivni faktor za razvoj infekcije Blastocistis hominisom, kao ni pohađanje kolektiva ili život u ruralnom sredinama. Stariji uzrast deteta (p=0,020) i život u kući (p = 0,033) su prediktivni faktori za pojavu hronične inflamatorne bolesti creva kod dece sa kolitisom. Deca sa blastocistozom su imala antropometrijske parametre u granicama normale.Ispitanici najčešće bili primljeni u bolnicu pod djagnozom gastroenterokolitisa, zbog proliva i bola u trbuhu, a da prisustvo gastrointestinalnih tegoba i prisustvo opštih znakova infekcije nisu jedan od sigurnih kliničkih značajnih znakova infekcije Blastocistis hominisom. Prisustvo patoloških primesa u stolici nije jedan od sigurnih klinički značajnih znakova infekcije Blastocistis hominisom.Na osnovu laboratorijskog, kliničkog i endoskopskog skora za aktivnost HIBC većina bolesnika je imala umerenu aktivnost.Inficirani sa Bh imaju najčešće C-reaktivni protein u okvirima refentnih vrednosti, izuzev ukoliko nemaju i HIBC. Povišena sedimentacija eritrocita je karakteristična za bolesnike sa HIBC. Oboleli od blastocistoze imaju najčesce imunoglobulin A, leukocite, neutrofile i eozinofile u krvi u referentnim granicama.Vrednosti feremije upućuju da je većina ispitanika bila anemična, a naročito deca koja su imala i infekciju sa Bh i HIBC. Kod bolesnika sa blastocistozom, postojanje pozitivnog testa na okultnu krv u stolici, treba da pobudi sumnju na udruženu HIBC. Ispitanici sa infekcijom Bh i sa HIBC su imali najčešće kvantitativno veći broj Bh u stolici. Mezenterajalni limfadenitis i splenomegalija su nespecifični ultrazvučni nalaz kod inficiranih sa Bh, iako su bili najčešće opisane patološke promene na ultrazvuku abdomena. Zaključujemo da su ispitanici najčešće imali nespecifične endoskopske promene i patohistološke promene u debelom crevu. Metronidazol je bezbedan i efikasan, u dozi 15-50 mg/kg/dan, u trajanju od 10 dana, u terapiji infekcije sa Bh kod dece. ZAKLJUČAK: Deca inficirana sa Bh imaju najčešće colitis od patoloških promena na debelom crevu, bez značajne razlike između nespecifičnog kolitisa i HIBC. Značajno manje inficiranih sa Bh ima uredan kolonoskopski nalaz.Utvrđivanja značaja Blastocistis hominisa u nastanku kolitisa i hronične inflamatorne bolesti creva kod dece, doprinosi prihvatanju Blastocistisa hominisa kao patogena i ukazuje na nephodnost njegovog lečenja.
INTRODUCTION: Blastocystis hominis (Bh) is the most outspread protist on our planet, but also the most controversial. Infection Bh starts by digestion of the eaten food or liquid which has been contained by a cyst form Bh. After swallowing, from the cyst they grow (progress) in the colon of the human, with a vacuolar form of a protest. Oral transmission is the most common way of spreading the infection. The disease caused by Bh on humans is called blastocystisis. In most cases the infected humans have gastrointestinal complaints, the most common are abdominal pain and diarrhea. Blastocystis is nowadays connected to two clinical disease, the irritable bowel syndrome and inflammatory bowel disease (IBD). THE AIM AND HYPOTHESESS: The subject of research is to establish the connection between the presence of the infection Bh and the existence of mucosal inflammation of the colon in children with gastrointestinal complaints, as well as to establish the group of the children with a special form of colitis, inflammatory bowel disease and the ones infected by Bh, wich would insure better understanding of the blastocystosis in children. The basic hypothesis in the study were statistically significantly higher incidence of chronic colitis and inflammatory bowel disease in children aged 1 month to 18 years, hospitalized for abdominal pain and/or diarrhea who are infected Bh, as well as significantly higher incidence of colitis compared in chronic inflammatory bowel disease in the same sample. MATERIALS AND METHODS: The prospective study included pediatric patients with abdominal pain and/or diarrhea, and stool positive on Bh, that have been hospitalized on the Department for gastroenterology, hepatology and nutrition, in the Institution for Health Care of Children and Youth in Vojvodina. The standard testing methods were used: anamnesis, physical examination, laboratory analysis of blood and stool, ultrasound examination of the abdomen, colonoscopy and histopathological examination of the biopsy of the colon. All patients have been treated with metronidazole for 10 days, according to the applicable protocols. RESULTS: The study included 102 patients, which are divided into three groups : 1. group (patients that have no colitis, included 4 patients (4.4%)), 2. group (patients with unspecified colitis, included 56 patients (56.55%)) and 3. group (patients with inflammatory bowel disease, included 42 patients (42.41%)). Among them, there was an equal number of children that were male and female, 51 boys and 51 girls. Age of respondents who have Bh infection ranged from 11 months to 17 years and 7 months. The median is 12.54 years, and the average age of 11.25 years. Blastocistosis no have seasonal character (χ2 = 0.667, df = 3, p = 0.881). Significantly more infected Blastocistis hominid lived in the house, but in an apartment owned and domestic animals and / or pets,yet the existence of appropriate hygiene, sanitation and water supply is not a predictive factor for the development of infection Bh, as well as attending the collective or life in rural areas . The older child's age (p = 0.020) and life at home (p = 0.033) were predictive factors for development of inflammatory bowel disease in children with colitis. Children with blastocistosis had anthropometric parameters within normal limits. Respondents most frequently been admitted to hospital under diagnosis gastroenteritis due to diarrhea and abdominal pain, and that the presence of gastrointestinal symptoms and general signs of infection are not a significant clinical signs of infection Bh. The presence of pathological findings in stool is not one of reliable signs of clinically infection Bh. Based on laboratory findings, clinical and endoscopic activity score for IBD most patients had moderate activity of desease. Children with Bh infection usually have normal C-reactive protein in terms of value, unless if have IBD. Elevated erythrocyte sedimentation rate is characteristic of patients with IBD. Children with blastocistosis usually have normal level of Immunoglobulin A, leukocytes, neutrophils and eosinophils. Serum iron indicate that most subject were anemic, especially children who have had an infection with the Bh and IBD.Children with blastocistosis, the existence of a positive test for occult blood in the stool, should arouse suspicion of association IBD. Subject with IBD had mostly quantitatively greater number of Bh in the stool. Mesenterial lymphadenitis and splenomegaly are non-specific ultrasound findings in infected with Bh, although they were usually described pathological changes in abdominal ultrasound. This is to conclude that the subject usually had colitis and IBD changes in endoscopic and histopathological changes in the colon. Metronidazole has beem proved safe and effective, at 15-50 mg/ kg/day for 10 days in the treatment of infections in children with Bh. CONCLUSION: Children infected with Bh colitis usually have pathological changes in the large intestine, with no significant difference between the non-specific colitis and inflammatory bowel disease. Significantly less infected with Bh has a normal colonoscopy findings. Confirmed the importance of Bh in the development of chronic colitis and inflammatory bowel disease in children, increase public acceptance Blastocistisa hominis as pathogens and points to the necessity of treatment.

Människor har olika riskuppfattningar beroende på tidigare erfarenheter, kulturell bakgrund och kunskap. Riskuppfattningarna kan skilja sig så att vissa människor upplever en inträffad händelse som en kris, medan andra upplever samma sak som en allvarlig händelse. Vid en kris eller allvarlig händelse prioriteras inte allmänhetens uppfattningar eller åsikter när ansvariga aktörer och myndigheter arbetar för att hitta orsaken till händelsen. Eftersom allmänheten är en stor grupp i samhället och är de som drabbas vid en kris eller allvarlig händelse, vill vi med denna studie undersöka hur de uppfattade det vattenburna Cryptosporidiumutbrottet i Östersund som pågick från november 2010 och tre månader framåt. För att genomföra studien har vi analyserat kvantitativa data från vår egen surveyundersökning som har genomförts med hjälp av enkäter. Den teoretiska referensram som ligger till grund för uppsatsen behandlar riskuppfattning samt kopplingar till risksamhället. De resultat som vi fick fram av studien visar att det finns signifikanta skillnader mellan ålder och hur man påverkades av Cryptosporidiumutbrottet, då i synnerlighet yngre påverkades mer av parasiten. Vidare finns stöd för detta resultat i tidigare forskning, som understödjer resonemanget att framförallt yngre personer påverkas mer av Cryptosporidium än äldre. Nyckelord: Vattenburen smitta, Cryptosporidium hominis, parasitutbrott, riskuppfattning och risksamhälle.

Robinson's xenic parasite culture method was used to investigate the prevalence of Dientamoeba fragilis and Blastocystis hominis in faecal samples submitted to the National Public Health Service for Wales (NPHS) laboratory at Aberystwyth. Culture, in combination with a commercial trichrome stain, gave a D. fragilis positivity-rate of 2.6% (25/976), and a B. hominis positivity-rate of 8.0% (78/976). Isolates of B. hominis and D. fragilis were typed using riboprinting of the amplified SSU rRNA gene. Sixty six B. hominis and thirty three D. fragilis isolates produced a product after amplification. The B. hominis typing results confirmed the extensive genetic variation reported by other workers, and six new ribodemes were found among the human isolates. Two animal Blastocystis isolates were typed; a monkey isolate was ribodeme 1, and an ovine isolate was a new ribotype. All D. fragilis isolates that were riboprinted were found to be genotype 1.

Objetivo: Determinar la prevalencia de infección por Strongyloides stercolaris y otras enteroparasitosis en el pueblo joven Santo Toribio de Mogrovejo de Chiclayo durante Junio-Octubre del 2011. Material y Métodos: Estudio descriptivo, trasversal; muestreo aleatorio, estratificado, polietápico, siendo el tamaño muestral de 106 pobladores. Se diseñó y validó una ficha de recolección epidemiológica. Un biólogo recibió entrenamiento en las técnicas de diagnóstico de Strongyloides stercolaris en un centro referencial de Lima. Se recolectaron 3 muestras por paciente, sometidas a 5 técnicas parasitológicas: Examen directo de heces, Baermann modificado en Copa por Lumbreras, Test de sedimentación espontánea, Cultivo en agar y Cultivo Dancescu. Resultados: Se visitaron 124 casas; el porcentaje de respuesta fue de 85,7%. Se logró entrevistar 106 personas. El promedio de edad fue de 27,8 +/- 16,9 años; hubieron 31 hombres (29,2%) y 75 mujeres (70,8%). El 26,4% de personas habían realizado un viaje a la Sierra y/o Selva en los últimos 5 años con una estancia mayor a un mes. El piso de tierra fue el más frecuente en el total de viviendas (55,6%); 102 personas (96,2 %) tenían desagüe; 23 pobladores (21,7 %) tuvieron al menos un parásito detectado. No se hallaron pobladores infectados con Strongyloides stercolaris. La enteroparasitosis más frecuente fue por protozoarios, con predominio de Blastocystis hominis en un 12,3%. Conclusiones: Se halló una baja frecuencia de enteroparasitosis y ausencia de pobladores infectados con Strongyloides stercolaris. El parásito más frecuente fue Blastocystis hominis.

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In tropical regions, resistance of cattle to external parasites is an important factor determining efficiency of the production systems. The objectives in this study were to evaluate the degree of resistance of Nelore (NE), Canchim x Nelore (CN), Aberdeen Angus x Nelore (AN) and Simmental x Nelore (SN) females of several ages to cattle tick (Rhipicephalus (Boophilus) microplus), horn fly (Haematobia irritans) and beef-worm (Dermatobia hominis), and the possibility of selection to increase resistance of beef cattle to these parasites, through the estimation of genetic parameters of the degree of natural infestation, in order to furnish information to beef cattle breeding programs in Brazil. Two experiments were carried out. In experiment 1, 16 NE, 18 CN, 16 AN and 16 SN 16.5 months old females, were artificially infested with 20,000 larvae of cattle tick, four times 14 days apart each, were done, and from day 18 to day 22 of each infestation the number of engorged female ticks (≥ 4.5 mm) was counted on the left side of each animal. In this experiment, data were analyzed as the percentage of return (PR = percentage of ticks counted relative to the number infested), transformed to (PR)1/4, using the least squares method with a model that included effects of genetic group (GG), animal within GG (error a), infestation number (I), GG x I, and the residual (error b). In experiment 2, from six to ten cattle tick, horn fly and beef-worm countings on NE (184), CN (153), AN (123) and SN (120) naturally infested females of several physiological states (calves, pregnant and open heifers, primiparous cows with and without a calf and pluriparous cows with and without a calf), from July 2003 to December 2004, were done. In this experiment, data, transformed to log10 (n + 1), were analyzed by the least squares method with a model that included effects of genetic group (GG) of female, animal within GG (error a), year-season of counting (YS), physiological state, and GG x YS interaction. Besides that, genetic parameters of the degree of infestation by the parasites were obtained by the restricted maximum likelihood method, using models that included fixed effects of contemporary group (genetic group, year-season of counting) and physiological state, and additive direct, animal s permanent environmental, and residual random effects. In artificial infestation, despite the GG x I interaction, in general, NE animals were more resistant, followed by CN, and, at last, by SN and AN ones, which showed, respectively, the following percentage of return: 0.35 ± 0.06, 0.54 ± 0.05, 0.89 ± 0.06 and 0.85 ± 0.06. In natural infestation, the difference among genetic groups depended on year-season of counting, however, in general, NE females were less infested by ticks than females of the other genetic groups, while AN females showed higher infestation by horn flies and beef-worm than females of the other genetic groups. Heritability and repeatability estimates based on one-trait analyses were 0.12 and 0.12, 0.30 and 0.30, and 0.04 and 0.12, for infestation by cattle ticks, horn flies and beef-worms, respectively, indicating that it is feasible to obtain genetic progress for infestation by horn flies. The genetic correlations among these traits were low, except that between infestations by horn flies and beef-worms (0.60).
Em regiões de clima tropical, a resistência dos bovinos a ectoparasitas é fator importante na determinação da eficiência dos sistemas de produção. Os objetivos neste trabalho foram avaliar o grau de resistência de fêmeas Nelore (NE) e cruzadas Canchim x Nelore (CN), Aberdeen Angus x Nelore (AN) e Simental x Nelore (SN) de diversas idades ao carrapato (Rhipicephalus (Boophilus) microplus), à mosca-dos-chifres (Haematobia irritans) e ao berne (Dermatobia hominis) e a possibilidade de seleção para aumentar a resistência de bovinos a esses parasitas, por meio da avaliação da existência de variação genética aditiva no grau de infestação natural, para fornecer subsídios aos programas de melhoramento genético de bovinos de corte do Brasil. Dois experimentos foram realizados. No experimento 1, 16 fêmeas NE, 18 CN, 16 AN e 16 SN, com média de idade de 16,5 meses, foram infestadas artificialmente com aproximadamente 20.000 larvas de carrapato, com intervalos de catorze dias, e do 18o ao 22o dia depois de cada infestação foram realizadas contagens de teleóginas semi-ingurgitadas (≥ 4,5 mm) do lado esquerdo do animal. Neste experimento, a percentagem de retorno (PR), ou seja, percentagem de carrapatos contados em relação ao total infestado, após transformação para (PR)1/4, foi analisada utilizando-se o método dos quadrados mínimos com um modelo que incluiu os efeitos de grupo genético (GG), animal dentro de GG (erro a), infestação (I) e GG x I, além do resíduo. No experimento 2, foram realizadas de seis a dez contagens de carrapatos, moscas-dos-chifres e bernes em fêmeas NE (184), CN (153), AN (123) e SN (120) de sete estádios fisiológicos, infestadas naturalmente, de julho de 2003 a dezembro de 2004. Neste experimento, os dados, transformados para log10 (n + 1), foram analisados pelo método dos quadrados mínimos com um modelo estatístico que incluiu os efeitos de grupo genético (GG) da fêmea, animal dentro de GG (erro a), ano-época da contagem (AE), estádio fisiológico e a interação GG x AE. Além disso, foram estimados parâmetros genéticos do grau de infestação por esses parasitas, pelo método da máxima verossimilhança restrita, com modelo que incluiu os efeitos fixos de grupo de contemporâneas (grupo genético, ano-época da contagem) e estádio fisiológico, além dos efeitos aleatórios aditivo direto, de ambiente permanente do animal e residual. Na infestação artificial, apesar da interação significativa entre grupo genético x infestação, em geral, os animais NE foram mais resistentes, seguidos do CN e, por último, dos SN e AN, apresentando, respectivamente, as seguintes médias de taxa de retorno: 0,35 ± 0,06; 0,54 ± 0,05; 0,89 ± 0,06 e 0,85 ± 0,06. Na infestação natural, a diferença entre os grupos genéticos dependeu do anoépoca da contagem, contudo, em geral, as fêmeas NE foram menos infestadas pelo carrapato do que as fêmeas dos outros grupos genéticos, enquanto as fêmeas AN foram mais infestadas pela mosca-dos-chifres e pelo berne do que as fêmeas dos outros grupos genéticos. As estimativas de herdabilidade e de repetibilidade foram 0,12 e 0,12; 0,30 e 0,30; e 0,04 e 0,12 para a infestação por carrapatos, moscasdos- chifres e bernes, respectivamente, indicando que há campo para seleção, principalmente para a infestação por moscas-dos-chifres. As correlações genéticas entre essas características foram baixas, exceto aquela entre as infestações por moscas-dos-chifres e por bernes (0,60).

Orientador: Ana Maria Lima de Azevedo Espin
Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia
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Blastocystis hominis es un enteropatógeno de distribución mundial en una variedad de animales, los cuales representan fuentes potenciales de infección para los humanos. El objetivo de este trabajo fue determinar el mejor medio de cultivo para el crecimiento de Blastocystis. Para ello se probaron tres medios de cultivo in vitro, Jones, Pavlova y Boeck-Drbohlav modificado. Se utilizaron 54 muestras de heces de cerdos previamente diagnosticadas como positivas a Blastocystis. El número de microorganismos inoculados se dividió en tres categorías: alto, intermedio y bajo. Cada muestra fue cultivada por duplicado en los tres medios y al cabo de 72 horas de incubación se procedió al recuento de microorganismos. Mediante un análisis de varianza se demostró que existe diferencia estadística significativa: 1) entre los medios evaluados, puesto que el medio Jones resultó ser el más eficiente, seguido de los medios Pavlova y Boeck-Drbohlav modificado y 2) entre las categorías del inóculo, donde se obtuvieron mejores resultados cuando el número de microorganismos inoculados se encontraban en la categoría bajo. Además, se pudo observar que la forma vacuolar fue la más comúnmente vista en los medios, seguida de la forma granular. Este estudio es un paso inicial importante para el desarrollo de técnicas más sofisticadas en la investigación de este microorganismo.

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

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With the objective to determine frequency, age and sex influences, and risk factor associated to Blastocystis hominis in feces of housed dogs and cats from the Metropolitan Region of Rio de Janeiro, Brazil, 234 fecal samples were collected by convenience from 175 dogs and 59 cats. To the diagnostic of B. hominis in fecal samples, direct examination was used, but ferric-hematoxilin and Gomori s trichrome techniques were used in order to confirm this diagnostic. Width and length of the parasite found in fecal samples varied from 10.07 to 13.80, and 12.66 to 19.93 to dogs and cats respectively. With regards the frequency of B. hominis in housed animals, 23.42 of dogs, and 23.72% of cats were positives, independent of animal sex. Animal s age was the important factor to determine, mainly in dogs, the risk of B. hominis transmission in dwellings.
Com o objetivo de determinar a morfologia, freq??ncia, influ?ncia da idade e sexo, e fatores de risco associados ? Blastocystis hominis nas fezes de c?es e gatos domiciliados na Regi?o Metropolitana do Rio de Janeiro, foram coletadas amostras fecais por conveni?ncia de 175 c?es e 59 gatos. Para o diagn?stico de B. hominis, foi utilizado o exame direto, e para confirma??o do diagn?stico foram usadas ?s t?cnicas de colora??o da hematoxilina f?rrica e tricr?mio de Gomori. A largura e o comprimento de B. hominis encontrado nas amostras fecais variaram de 10,07 a 13,80μm, e 12,66 a 19,93μm para c?es e gatos respectivamente. Quanto ? freq??ncia de B. hominis nos animais domiciliados, 23,42% dos c?es, e 23,72% dos gatos foram positivos, independente do sexo. A idade dos animais foi um importante fator para determinar, principalmente nos c?es domiciliados, o risco de transmiss?o de B. hominis.

Las enteroparasitosis intestinales constituyen un problema de salud pública en Perú, debido a que estos parásitos pueden ingresar al organismo por vía oral y hábitos higiénico-sanitarios deficientes que facilitan su transmisión y conservación. Cuando la carga de dichos parásitos es considerablemente alta o se acompaña de alteraciones en la inmunidad del hospedero, se pueden producir complicaciones que comprometen seriamente la salud del paciente. Sabemos que el control farmacológico de las parasitosis es efectivo y seguro. No obstante, sin autocuidado y mantenimiento sostenible de buenas condiciones higiénico-sanitarias, no es posible su erradicación. Considerando lo mencionado anteriormente, desarrollaré la presente investigación en una población escolar de la Institución Educativa N° 6041 “Alfonso Ugarte” del distrito San Juan de Miraflores para determinar la prevalencia de parasitosis intestinales. Se eligió una muestra representativa conformada de 116 niños de 8 a 13 años. Las muestras fecales obtenidas fueron analizadas utilizando: examen macroscópico, método directo, método de Parodi Alcaraz y test de Graham. El 85.3% de los alumnos examinados resultaron parasitados. La incidencia parasitaria fue mayor en mujeres (86.8%) comparado a los hombres (83.6%). La frecuencia parasitaria de acuerdo al Monoparasitismo de los grupos taxonómicos fueron 35.3% del Phylum Amoebozoa, 3.4% del Phylum Metamonada, 3.4% del Phylum Platyhelminthes, 0.9% del Phylum Bigyra y 0.9% del Phylum Nematoda, con las especies Entamoeba coli, Giardia lamblia, Hymenolepis nana, Blastocystis hominis y Enterobius vermicularis, respectivamente. La mayor frecuencia correspondiente al Biparasitismo fue la asociación de los Phyla Metamonada y Amoebozoa con 32.8%. La mayor frecuencia correspondiente al Triparasitismo fue la asociación de los Phyla Metamonada, Amoebozoa y Platyhelminthes con 1.7%. The intestinal enteroparasites constitute a public health problem in Peru, due to these parasites can enter the body by mouth and hygienic habits-poor health that facilitate its transmission and conservation. When the burden of such parasites is considerably high or is accompanied by alterations in the immunity of the host, it can produce complications which seriously compromise the health of the patient. We know that the pharmacological control of the parasitosis is effective and safe. However, self-care and sustainable maintenance of good hygienic and sanitary conditions, it is not possible to its eradication. Considering the above, I will develop this research in a school population of the Educational Institution N° 6041 "Alfonso Ugarte" of the district San Juan de Miraflores to determine the prevalence of intestinal parasitism. I chosen a representative sample consisted of 116 children 8 to 13 years old. Stool samples obtained were analyzed using: macroscopic examination, direct method, Parodi Alcaraz’s method and Graham’s test. The 85.3% of the students examined were parasitized. The parasitic incidence was higher in women (86.8%) compared to men (83.6%). The frequency of parasites according the Monoparasitism of taxonomic group was 35.3% of the Phylum Amoebozoa, 3.4% of the Phylum Metamonada, 3.4% of the Phylum Platyhelminthes, 0.9% of the Phylum Bigyra and 0.9% of the Phylum Nematoda, with the species Entamoeba coli, Giardia lamblia, Hymenolepis nana, Blastocystis hominis and Enterobius vermicularis, respectively. The highest frequency corresponding the Biparasitism was the association of the Phyla Metamonada and Amoebozoa with 32.8%. The highest frequency corresponding the Triparasitismo was the association of the Phyla Metamonada, Amoebozoa and Platyhelminthes with 1.7%.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Avaliou-se comparativamente duas formulações, de ação prolongada, contendo Ivermectina (4% e 3,15%) em quatro experimentos em bovinos. Pelo grupo controle, constatou-se a presença das seguintes espécies de nematódeos gastrintestinais: Haemonchus placei, Cooperia punctata, Cooperia spatulata, Trichostrongylus axei, Oesophagostomum radiatum e Trichuris discolor. Quanto à eficácia anti-helmíntica, observou-se que a Ivermectina 4%* foi estatisticamente (P<0,05) superior à Ivermectina 3,15% contra Haemonchus placei, Cooperia punctata e Oesophagostomum radiatum. As duas formulações praticamente não diferiram quanto à eficácia anti-Boophilus microplus e antilarvas de Dermatobia hominis (berne). Decorridos 120 dias pós-tratamento, foram registrados, em relação ao grupo controle, diferenciais de ganho de peso corporal de 17,79 e 11,19 kg em bovinos medicados com Ivermectina 4% e Ivermectina 3,15%, respectivamente.
Comparative endectocide efficacy and effect on weight gain in cattle treated with two (3.15% or 4%) ivermectin formulations. Four trials were conduced to a comparative evaluation of two long action (3.15% or 4%) ivermectin formulations. In the anthelmintic study, the 4% ivermectin formulation showed efficacy statistically higher than 3.15% ivermectin against Haemonchus placei, Cooperia punctata and Oesophagostomum radiatum. Ivermectin at both concentrations was similarly effective against Boophilus microplus and larvae of Dermatobia hominis (warble fly). After 120 days post-treatment, it was observed, relative to the control group, difference in the weight gain of 17.70kg and 11.10kg in cattle treated with 4% ivermectin and 3.15% ivermectin, respectively.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Os probióticos são microorganismos vivos que afetam de forma benéfica o hospedeiro ou o ambiente. Na aquicultura podem ser usados tanto na água como na ração, mas seu uso na alimentação é destacado como uma das principais medidas profiláticas. As doenças bacterianas são consideradas um dos principais entraves no crescimento da aquicultura, e assim, há a necessidade urgente no desenvolvimento de probióticos. O tambaqui Colossoma macropomum é a espécie nativa mais produzida no Brasil, e apesar de sua importância econômica, não há estudos que estabeleçam os microorganismos com potencial probiótico para esta espécie de peixe. Neste trabalho foi possível identificar e caracterizar bactérias autóctones com potencial probiótico para o tambaqui a partir de testes de: caracterização morfológica, catalase, tolerância à bile, antagonismo frente à patógenos, sensibilidade a antimicrobianos e sequenciamento do gene 16S rRNA. As cepas selecionadas foram: Enterococcus faecalis, Enterococcus hirae, Lactococcus lactis, Pediococcus pentosaceus, Staphylococcus hominis e Staphylococcus saprophyticus. Todas as cepas foram tolerantes aos ácidos biliares do tambaqui e capazes de inibir o crescimento dos patógenos Enterococcus casseliflavus, Lactococcus garvieae e Aeromonas hydrophila. Todas as cepas foram parcialmente resistentes contra sete antibióticos. Como as cepas de S. saprophyticus e E. faecalis apresentaram menores valores no teste de antagonismo e por estas bactérias serem relatadas como agentes zoonóticos, concluímos este estudo selecionando quatro potenciais cepas: E. hirae, L. lactis, P. pentosaceus, S. hominis. Este é o primeiro estudo a referir o potencial uso probiótico de cepas autóctones para o tambaqui.
Probiotics are living microorganisms that beneficially affect the host or the environment. In aquaculture it can be used in both water and feed, but its use in feed is highlighted as one of the main prophylactic measures. Bacterial diseases are considered to be one of the major obstacles to the growth of aquaculture, and thus, there is an urgent need for the development of probiotics. The tambaqui Colossoma macropomum is the most produced native species in Brazil, and despite its economic importance, there are no studies that establish the microorganisms with probiotic potential for this fish species. In this study it was possible to identify and characterize autochthones bacteria with probiotic potential for tambaqui from tests of: morphological characterization, catalase, bile tolerance, antagonism of pathogens, antimicrobial susceptibility and 16S rRNA gene sequencing. The selected strains were: Enterococcus faecalis, Enterococcus hirae, Lactococcus lactis, Pediococcus pentosaceus, Staphylococcus hominis and Staphylococcus saprophyticus. All strains were tolerant to acids bile from tambaqui and capable of inhibiting the growth of Enterococcus casseliflavus, Lactococcus garvieae and Aeromonas hydrophila pathogens. All strains were partially resistant against seven antibiotics. Since the strains of S. saprophyticus and E. faecalis presented lower values in the test of antagonism and because these bacteria were reported as zoonotic agents, we conclude this study selecting four potential strains: E. hirae, L. lactis, P. pentosaceus, S. hominis. This is the first study to mention the potential probiotic use of autochthonous strains for tambaqui.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Universidade Estadual Paulista (UNESP)
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.
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.

Orientador: Ana Maria L. de Azeredo Espin
Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia
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Mestrado