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Čerstvé a měkké sýry jako potenciální zdroj \kur{Encephalitozoon cuniculi}
VECKOVÁ, Tereza
The aim of this work was to determine the occurrence and prevalence of Encephalitozoon cuniculi in goat's milk and faeces, and to evaluate the effect of pasteurization and curdling in goat cheese production on the infectivity of E. cuniculi spores for immunodeficient (SCID, CD4-/- and CD8-/-) and immunocompetent (BALB / ca C57BL/6) mice. Under experimental conditions, spores of E. cuniculi genotype II (557,000 spores in 1 g of cheese) remain viable in fresh cheeses treated by pasteurization at 72 ° C for 20 seconds and are able to cause infection in laboratory animals. Using nested PCR, specific DNA of E. cuniculi genotype I and genotype II was detected in eight of the nine goat cheeses purchased from various producers/breeders in the Czech Republic in the amount of 1 to 202 spores per 1 g of cheese. These cheeses were fed to immunodeficient and immunocompetent mice at a dose of 60 g/mouse within 24 days. The results of the experiment showed that spores of E. cuniculi genotype I and II remain viable in both fresh and soft cheeses and are able to cause infection in laboratory animals. Based on the results obtained, fresh and soft cheeses should be considered as a potential source of E. cuniculi infection for humans.
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Fermentované masné výrobky jako potenciální zdroj Encephalitozoon cuniculi
VECKOVÁ, Tereza
This study describes the prevalence and burden of Encephalitozoon cuniculi in pork meat and evaluates the effect of fermentation in the production of sausages on E. cuniculi infectivity for immunedeficient (SCID) and immunocompetent (BALB/c and C57BL/6) mice. Using a nested polymerase chain reaction approach, 2 of 50 slaughter pigs was found to have E. cuniculi genotype II in meat in an amount 60-250 spores per 1 g. Under experimental conditions, E. cuniculi genotype II spores (3000 spores per 1 g) in meat remained infective for mice following fermentation treatments at 24 °C for 48 hrs and consumed within two weeks after production. Based on these findings, fermented meat products should be considered as a potential source of E. cuniculi infection in humans.
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Milk as potencial source Encephalitozoon cuniculi
TOMANOVÁ, Vendula
The object of this thesis is to find out an incidence and prevalence of Microsporidia infection in uncooked cow´s milk. Samples of milk, faeces and urine from 147 milk cows were screened. One of them was positive for presence of Encephalitozoon cuniculi in sample of milk and stool repeatedly. Theoretic part includes information about Microsporidia, e.g. developmental cycle, transfer and course of disease, treatment and prevention. There is a description of heat treatment of milk as well. A methodical procedure of microsporidia DNA detection from samples of milk, feaces or urine is described in the practical part. The thesis is focused on experimental verification of sterilizing capabilities of various pasteurization methods. Milk was contaminated by Encephalitozoon cuniculi and then heat treatment according to the methods used in practice followed. Contaminated milk was offered to laboratory immunodeficient SCID mice. After three weeks mice were put to death and their organs were examined for the presence of Microsporidia DNA using molecular methods. Finding out of Microsporidia in examined organs was the evidence of insuffient heat treatement.
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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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Endoparasites of horses
WAGNEROVÁ, Pavla
A total of 587 faecal samples were collected from 98 horses of different age from three farms with various breeding system (pasture, stable and combination) in the South Bohemia, Czech Republic during three consecutive years (from 2009 to 2011), and screened for the presence of endoparasites, especially these inhabiting gastrointestinal tract and lung, using standard parasitological methods including flotation, sedimentation and staining methods. Moreover presence of Encephalitozoon spp. was detected using genus specific nested PCR. Small strongyles were the most common nematodes among studied horses. The age of animals was evaluated as a risk factor in relation to Eimeria leuckarti, Strongyloides westeri and Parascaris equorum infection in foals. The resistance of small strongyles (Cyathostominae) to benzimidazol anthelmintics has been revealed only on one of the visited farm. Most of examined animals were mono-infected. The breeding of horses in the stable represent management system with lowest risk in relation to parasitic infection of animals. Molecular analysis revealed the presence of human pathogenic Encephalitozoon cuniculi genotype I in horses.
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The course of microsporidiosis caused by \kur{Encephalitozoon cuniculi} in immunocompetent and immunodeficient mice
KOTKOVÁ, Michaela
The course of microsporidiosis caused by Encephalitozoon cuniculi in immunocompetent BALB/c mice and immunodeficient SCID mice was screened using molecular methods. The site of infection in organs was located using molecular and histology methods. The effectiveness of albendazole treatement and possibility of infection relapse after immunosuppresion (cyclosporine A, tacrolimus, mycofenolate mofetil) was also studied. Moreover, the course of excretion of microsporidial spores in feces was monitored during the whole time of experiment.
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The influence of interferon gamma and specific antibodies on the p.o. infection with \kur{Encephalitozoon cuniculi in vivo}
JELÍNEK, Jiří
The influence of interferon gamma and specific antibodies on the infection with E. cuniculi in vivo has been studied. Reconstruction of SCID mice with CD4+ T-lymphocytes from BALB/c mice and from mice with defect gene for interferon gamma was used. Effects of the treatment with mouse recombinant interferon gamma and anti-E. cuniculi sera on survival of E. cuniculi infected SCID mice were monitored. The influence of the immunization with E. cuniculi antigen on the survival of E. cuniculi infected mice with defect gene for interferon gamma was examined.
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