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The course of microsporidiosis caused by Encephalitozoon cuniculi in experimental model
VOTOČKOVÁ, Tereza
Microsporidia are obligate intracellular parasites causing disease called microsporidiosis. The infectious strategy is a spore - a highly organized cell which gives a rise to the infection of the host. This bachelor thesis records the progress of microsporidiosis provoked by the infection of the experimental model of the species of microsporidia Encephalitozoon cuniculi, namely of the genotype ECIII. The theoretical part includes the familiarization with microsporidia. In this part, there are first records leading to paying more attention to them and initiating their closer examination. This section describes both the biological characteristics and the systematic categorization of microsporidia and the description of a spore as the infectious stage of life cycle, which enables get into the host body where microsporidia reproduce and cause diseases. The sources of microsporidia are mentioned as well as the ways of transmission and microsporidia species most commonly detected in human whose attention is devoted mainly to Encephalitozoon cuniculi - its history and development, the infectious progress in hosts, the detection, and the treatment. The practical part describes the methodical process of the way of monitoring of microsporidiosis on experimental models in a form of laboratory mice of inbred immunocompetent BALB/c and the mice with severe combined immunodeficiency SCID. The mice were orally infected with spores of Encephalitozoon cuniculi genotype ECIII, which was isolated from steppe lemming (Lagurus lagurus). The progress of infection was monitored both on the basis of coprological examination of mouse feces and on the basis of the examination of tissue and body fluid samples. The processing of individual samples was carried out with the aid of molecular diagnostics in the parasitological laboratory AV ČR in České Budějovice. The DNA from individual tissues and body fluids was isolated by using a commercially supplied kit, QIAamp DNeasy Blood & Tissue Kit ( QIAGEN), the feces were treated with a commercially supplied QIAamp DNA Stool Mini Kit (QIAGEN). The principle was to destroy the individual spores and to obtain the pure DNA which was suitable for further processing. Subsequently, the two-step polymerase chain reaction was carried out. providing rapid multiplication of DNA. The next step was a gel electrophoresis which arouses the division of the DNA into individual fragments on the basis of different molecular weights due to the effect of unidirectional electrical current. The fragments were visualized by UV transilluminator at a wavelength of 312 nm linked to a computer. The part of the research was the albendazole treatment of BALB/c mice. The drug was orally administered in the range from 28th to 42nd day after the infection. The results on progress of microsporidiosis through various organs and tissues, including the involved treatment, were recorded in well arranged tables. At the end of this bachelor thesis, there is a summary of the research and its comparison with previous studies. The results showed that microsporidia may be a threat not only for people suffering from immune deficiency, but thanks to their successful survival in the organs of immunocompetent hosts and their ability of activation from the undetectable level, they emphasize the danger of latent microsporidiosis as a risk and life-threatening factor for groups of people undergoing chemotherapy or organ transplantation where the patients can receive inflected graft from a donor.
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