Orientador: Regina Maura Bueno Franco / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-03T20:53:37Z (GMT). No. of bitstreams: 1 CantusioNeto_Romeu_M.pdf: 1625548 bytes, checksum: e91f470bbd9fce0e7bb535b92359fbac (MD5) Previous issue date: 2004 / Mestrado

Orientador: Regina Maura Bueno Franco / Dissertação (mestrado) - Universidade Estadual de Campinas. Instituto de Biologia / Made available in DSpace on 2018-08-04T03:37:16Z (GMT). No. of bitstreams: 1 Oliveira_PaulaVilmade_M.pdf: 3163607 bytes, checksum: cf552ae60ce22ed636ee5cbfa344ebd8 (MD5) Previous issue date: 2005 / Resumo: Giardia spp. e Cryptosporidium spp. estão amplamente dispersos no ambiente aquático e foram responsáveis por diversos surtos de giardiose e criptosporidiose por veiculação hídrica em vários países. A presença destes patógenos em água utilizada para consumo humano é um fator de risco para aquisição destas parasitoses porque cistos e oocistos são resistentes aos processos de tratamento e às condições ambientais. Deste modo, a preocupação em relação à contaminação das fontes de água para uso humano por estes protozoários é crescente. No Brasil, há registros da ocorrência de ambos organismos em água superficial e subterrânea e em esgoto bruto e tratado. Atualmente, há uma recomendação oficial para a implantação da pesquisa de cistos e oocistos em água tratada. Assim, é necessário ter conhecimento sobre a freqüência e a distribuição destes parasitos nos mananciais. Os objetivos do presente estudo foram: i) verificar a ocorrência de cistos de Giardia e oocistos de Cryptosporidium em amostras de água bruta superficial do Rio Atibaia (Campinas, São Paulo) utilizando a técnica de filtração em membrana (47 mm diâmetro; 3 ?m de porosidade nominal); ii) avaliar a influência do desaguamento do Ribeirão Pinheiros na qualidade da água do Rio Atibaia mediante a comparação e a correlação entre parâmetros microbiológicos, físicos, pluviométricos e concentração de cistos e oocistos em dois pontos: à montante (A1) e à jusante (A2) do desaguamento do Ribeirão Pinheiros; e iii) verificar a ocorrência de variação sazonal destes protozoários na água superficial do Rio Atibaia. De março de 2002 a abril de 2003, 50 amostras de água foram analisadas. No ponto A1, cistos de Giardia foram detectados em 52,0% das amostras (concentração = 25,12 cistos/l), enquanto no ponto A2, a positividade foi de 76,0% (64,16 cistos/l). Oocistos de Cryptosporidium foram detectados somente no ponto A2, em 0,04% das amostras (0,8 oocistos/l). Houve diferença significativa na concentração de cistos entre os pontos A1 e A2 (p = 0,011) e correlação entre a pluviosidade e os parâmetros: concentração de cistos de Giardia (r = 0,611); coliformes totais (r = 0,467) e fecais (r = 0,467) no ponto A2. Em ambos os pontos, não foi encontrada correlação entre concentração de cistos de Giardia e as variáveis: coliformes totais (A1: r = -0,276; A2; r = -0,201), fecais (A1: r = -0,187; A2: r = -0,201) e turbidez (A1: r = -0,252; A2: r = -0,055). Dentre estes parâmetros, apenas a turbidez teve variação entre verão/inverno e verão/outono (p = 0,06). A técnica de filtração em membrana apresentou uma eficiência de recuperação de 72,2% para cistos de Giardia e de 65,1% para oocistos de Cryptosporidium. Os resultados obtidos neste estudo são relevantes porque o Rio Atibaia é o manancial que abastece a cidade de Campinas e outros municípios da região e o conhecimento da epidemiologia ambiental destes protozoários é importante do ponto de vista da Saúde Pública e também das companhias de tratamento de água / Abstract: Giardia spp. and Cryptosporidium spp. are widely dispersed in the aquatic environment and are responsible for several waterborne outbreaks of giardiosis and cryptosporidiosis in many countries. The occurrence of these pathogens in drinking water is a risk factor for human infection because of the resistance of cyst and oocyst to water treatment processes and to the environment. Nowadays, there is an increasing concern about drinking water contamination by these protozoans. In Brazil, the presence of both organisms was registered in groundwater and superficial water and in raw and treated sewage. Currently, there is an official recommendation for monitoring the presence of Giardia and Cryptosporidium in drinking water samples; thus a federal decree aims to establish researches for (oo)cysts detection in treated water. The knowledge about the frequency and distribution of these protozoan parasites in water sources is necessary to establish policies on public health. The aims of the present study are: i) to verify the occurrence of Giardia cysts and Cryptosporidium oocysts in Atibaia River¿s (Campinas City, São Paulo State) superficial raw water samples using a membrane filtration method (47 mm diameter; 3 ?m of nominal porosity; ii) to evaluate the influence of the discharge of Pinheiros Stream on the water quality of the Atibaia River by the correlationship among microbiological and physical parameters, rainfall and oocysts levels in two points: upstream (A1) and downstream (A2) of Pinheiros Stream; and iii) to verify the seasonal variation of these protozoa occurrence in the superficial water of Atibaia River. From March 2002 to April 2003, 50 water samples were collected. In point A1, Giardia cysts were detected in 52,0% of the samples (concentration = 25,12 cysts/l) while in point A2 the positivity for this parasite was 76,0% (concentration = 64,16 cysts/l). Cryptosporidium oocysts were detected only in A2 point, in 0,04% of the samples (concentration = 0,8 oocysts/l). There was a significant difference for cysts concentration between A1 and A2 points (p = 0,011) and among rainfall and the parameters: cyst concentration (r = 0,611); total (r = 0,467) and fecal coliforms (r = 0,467) in A2 point. In both points, it was not found a correlationship among cyst concentration and the following parameters: total coliforms (A1: r = -0,276; A2; r = -0,201), fecal coliforms (A1: r = -0,187; A2: r = -0,201) and turbidity (A1: r = -0,252; A2: r = -0,055). The turbidity showed a variation between summer and winter periods as well as summer and fall periods (p = 0,06). The membrane filtration technique attained recovery efficiency rates of 72,2% for Giardia cysts and 65,1% for Cryptosporidium oocysts. The data obtained in this study are relevant because the Atibaia River is the major water source found in Campinas and other cities in the region. Thus, the knowledge about the environmental epidemiology of these protozoa is of very important to policies on to the Public Health and to the drinking water industry / Mestrado / Parasitologia / Mestre em Parasitologia

Intestinal infections with pathogenic protozoa may cause severe disease and remain a neglected problem in regions with inadequate sanitation and hygiene standards, especially in sub-Saharan Africa. Unfortunately, very little data about the prevalence of these infections in risk groups exist from the region. The present study was therefore conducted to assess the prevalence of intestinal protozoan infections in a representative population sample on Ijinga Island, north-western Tanzania. Methods: This was a cross-sectional study which was carried out in 2016 as part of the on-going SchistoControl pilot project on Ijinga Island, north-western Tanzania. A single stool sample was collected from 357 participants and examined microscopically for presence of trophozoites or cysts of intestinal protozoan parasites. In addition, real-time polymerase chain reaction (qPCR) was used to determine the species of intestinal protozoa. Results: Based on microscopy and qPCR, the prevalence of Giardia intestinalis infection was 12% and 15.1%, respectively. Based on microscopy, the prevalence of Entamoeba histolytica/Entamoeba dispar was 26.1%. However, through species identification using qPCR, 21.8% of the study participants were carrying non-pathogenic E. dispar and none of them was infected with E. histolytica. Conclusion: Intestinal protozoan infections are common among the population in the study area. The detection of these infections in different age groups indicates a poor hygienic standard in the community. Improvement in water, sanitation, hygiene and public health education on hand washing will help in controlling these infections. / Intestinale Infektionen mit pathogenen Protozoen können schwere Erkrankungsbilder verursachen und stellen eine oftmals vernachlässigte Problematik, vor allem in Regionen mit geringen Sanitär- und Hygienestandards, dar. Dies betrifft insbesondere südlich der Sahara gelegene Länder. Hier existieren leider keine oder nur unzureichende Prävalenzdaten, was zum Teil auch auf die inadäquaten diagnostischen Möglichkeiten zurückzuführen ist. Die Übertragung erfolgt fäkal-oral über verunreinigte Nahrungsmittel sowie kontaminiertes Trinkwasser und kann symptomatische und asymptomatische, akute und chronische Infektionen hervorrufen. Methoden: Es handelt sich um eine Querschnittsstudie, die 2016 im Rahmen des laufenden SchistoControl-Pilotprojekts auf der Insel Ijinga im Nordwesten Tansanias durchgeführt wurde. Es wurden Stuhlproben von 357 Teilnehmern gesammelt und mikroskopisch auf Trophozoiten oder Zysten von intestinalen Protozoen untersucht. Zusätzlich wurde die Realtime-Polymerasekettenreaktion (qPCR) zur Speziesdifferenzierung verwendet. Ergebnisse: Basierend auf Mikroskopie und qPCR betrug die Prävalenz von Giardia intestinalis 12% bzw. 15,1%. Basierend auf der Mikroskopie betrug die Prävalenz von Entamoeba histolytica / Entamoeba dispar 26,1%. Durch Speziesdifferenzierung mittels qPCR waren jedoch 21,8% der Studienteilnehmer positiv für E. dispar und keiner positiv für E. histolytica. Schlussfolgerung: Intestinale Protozoeninfektionen sind in der Bevölkerung des Studienortes häufig. Der Nachweis dieser Infektionen in verschiedenen Altersgruppen weist auf einen geringen Hygienestandard in der Gemeinde hin. Eine Verbesserung der Wasser-, Sanitär-, Hygienestruktur und des Gesundheitsbewusstseins wird zur Kontrolle dieser Infektionen beitragen.

Se determina la prevalencia de Giardia spp. en ratas urbanas “Rattus rattus” y “Rattus norvegicus” capturadas en un zoológico de Lima Metropolitana, además , se evaluó la asociación del parasito con las variables especie, sexo y edad de estos roedores. Se atraparon 127 roedores (Rattus spp.), usando trampas especiales (tipo“Tomahawk”). Se siguió un estricto control de bioseguridad para la manipulación y recolección de muestras de estos animales, sirviendo de referencia los protocolos proporcionados por el “Centro de Enfermedades Infecciosas y Prevención de Atlanta (CDC)”. Los datos que se tomaron en cuenta para este estudio fueron especie, sexo y edad de los roedores, y las muestras recolectadas provinieron del recto. La conservación se hizo utilizando formol al 10% y posteriormente fueron procesadas en el Laboratorio de Parasitología de la FMV-UNMSM. La técnica empleada para determinar la presencia de Giardia spp. fue la de Ritchie modificado, siendo muestra positiva aquella en que se hallara la presencia de quiste o trofozoito de Giardia spp. Como resultado se halló una prevalencia de Giardia spp. de 5.5 ± 0.04% (7/127); y se hayo una prevalencia del 2.1% del genero Rattus rattus (1/48) y 7.6% en Rattus norvergicus (6/79). Mediante la prueba de Chi cuadrado se encontró que No existe asociación con una significancia de “(p > 0,05)” entre la presencia de Giardia spp. y las variables especie, sexo y edad, por lo que la prevalencia de Giardia spp. en ratas urbanas en el presente estudio, es baja. Sin embargo, es de consideración tomar en cuenta la posibilidad de transmisión de enfermedades parasitarias de este animal a una población vulnerable como son niños y ancianos, trabajadores y animales del zoológico. / Tesis

Advances in DNA sequencing led to a technological breakthrough, that allowed analyzis of complete genomes including those of parasitic protists Trichomonas vaginalis and Giardia intestinalis . These organisms are studied not only for their clinical importance, but also from the evolutionary point of view for their adaptation to anaerobic environment. Genome sequencing and annotations of predicted proteins alone did not bring detail view into functioning of their mitochondrion related organelles ­ in G. intestinalis mitosomes, not­participating in energetic metabolism, in T. vaginalis hydrogenosomes, producing molecular hydrogen and ATP by means of substrate phosphorylation. Traditional methods based on a fractionation by ultracentrifuging in density gradient and subsequent biochemical and enzymological analyzes were extended by one­ and two­dimensional electrophoresis with subsequent identification of proteins by mass spectrometry. Methods of multidimensional separation of peptides produced by specific proteolysis of a complex mixture...

7 ABSTRACT Mitochondria of opisthokonts undergo permanent fusion and fission throughout the cell cycle. Keeping these two processes in balance is vital for various aspects of mitochondrial and cellular homeostasis. Both mitochondrial fusion and division mechanisms are controlled by highly conserved dynamin-related GTPases that are present in all kingdoms of life. The aspects of mitochondrial dynamics outside the opisthokonts is, however, almost completely unexplored phenomenon. In our work, we introduced a tool for live imaging of the reduced forms of mitochondria into model organisms Giardia intestinalis and Trichomonas vaginalis, anaerobic protist parasites from the Excavata supergroup of Eukaryotes. Using this technique, we investigated the dynamics of the mitosomes, the simplest forms of mitochondria, of G. intestinalis. The division of mitosomes is restricted to Giardia mitosis and is absolutely synchronized with the process. The synchrony of the nuclear and the mitosomal division persists also during the encystation of the parasite. Surprisingly, the sole dynamin-related protein of the parasite seems not to be involved in mitosomal division. However, throughout the cell cycle mitosomes associate with the...

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Cats are infected with a variety of internal parasites, some of which are zoonotic. Therefore, being able to effectively detect and determine prevalence of internal parasites in cats is important for both feline and human health. Some parasites are easier to detect than others. Diagnosing Giardia duodenalis and Cryptosporidium spp. can be difficult because cysts and oocysts shed in the feces are small, shed intermittently, and require a trained technician to consistently identify them. As a result, infections with these protozoan parasites can be missed. Fecal immunoassays detect antigens in feces and can have increased sensitivity when compared to traditional microscopic techniques, but still do not detect every infection. The current reference standard is an immunoassay known as the direct immunofluorescent assay, but it requires expensive equipment and a long incubation period. As a result, two prototype lateral flow fecal immunoassays, the Cryptosporidium EZ VUE and Giardia EZ VUE, designed by TECHLAB® Inc were evaluated for the ability to detect G. duodenalis and Cryptosporidium spp. infections in cats because they are cheap, easy to use, easy to store and easy to interpret. In addition, samples were examined using a 33% zinc sulfate (ZnSO4) centrifugal fecal flotation procedure and the MERIFLUOR® Cryptosporidium/Giardia direct immunofluorescent assay (IFA), which served as the reference test. Other internal parasites found on the centrifugal fecal flotation with zinc sulfate were recorded to determine prevalence. Both EZ VUE fecal immunoassays demonstrated potential in diagnosing infections in cats when compared to centrifugal fecal flotation and the reference. Additionally, a variety of other internal parasites were identified. This included several potentially zoonotic species including Spirometra mansonoides, Ancylostoma sp. and Toxocara cati, which was also the most commonly identified species of parasite. Additionally, it was determined that several factors may contribute to higher prevalence of parasites especially in cats with the status of stray or feral. / M. S. / Internal parasites affect both human and animals. Some parasites do not cause any clinical signs or have a lasting effect, while others can cause damage or contribute to the death of an organism. Certain parasites are zoonotic, meaning they can be transmitted between humans and animals. Several of these zoonotic parasites are found in companion animals, such as cats. This is especially important because cats are a common household pet. How owned animals are treated, especially pets like dogs and cats, has changed over the past few decades. In many of these households they are considered family members. Additionally, there is a large population of free roaming cats, both stray and feral, that have the potential to interact with humans or their pets. Therefore, it is important to be able to effectively diagnose and determine the prevalence of parasites for both feline and human health. Some parasites are harder to diagnose than others due to varying factors and more diagnostic tests are needed to effectively and efficiently detect them. Two of these parasites that can be challenging to detect are Giardia duodenalis and Cryptosporidium spp. This thesis was aimed at evaluating two prototype diagnostic tests, originally designed for use in humans, for ability to detect these parasites and their potential as diagnostic tests in cats. In addition, this thesis determined the prevalence of other internal parasites found in a large group of cats in Virginia. The obtained results indicated the prototype diagnostic tests had potential. A variety of internal parasites were also present in Virginia cats, some of which were zoonotic, and showed how important routine veterinary care for maintaining feline and human health