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Tažní psi v Arktidě jako potenciální zdroj parazitárních infekcí lidí a volně žijících zvířat
BROŽ, Marek
The aim of this thesis is to evaluate dogs introduced into the Arctic as a source of intestinal parasites infection both for wildlife and humans. To that aim coprological examination of faeces sampled in Svalbard and Greenland was performed. Microscopical and molecular detection of cryptosporidia, giardiae, microsporidia, roundworms and tapeworms in faeces was carried out. One sample from Svalbard was positive for Toxocara canis eggs and four samples from Svalbard were positive for unusual genotype of Enterocytozoon bieneusi.
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Structure and function of the invasion apparatus of microsporidia
Dohnálek, Vít ; Doležal, Pavel (advisor) ; Vávra, Jiří (referee)
Microsporidia are obligate intracellular parasites that cause significant economic damage and can infect humans. They had evolved highly specialized invasion apparatus that is unique to them. During the invasion the sporoplasm is released from the spore wall and it is transferred to the host cytoplasm through the polar tube. Current research on the invasion apparatus has been focused mainly on the polar tube that is its most prominent structure. Polaroplast and posterior vacuole remain out of the main interest, although they are necessary for the activation and execution of the invasion. If the right combination of environmental factors occurs, the organization of the polaroplast begins to change, the posterior vacuole starts to swell and the polar tube is discharged. Sporoplasm is eventually pushed through the tube into the host cell by growing posterior vacuole. The mechanism has not been explained yet however plenty of theories are trying to explain the germination. This work summarizes current theories and knowledge of structure and function of particular parts of the invasion apparatus. Key words: Microsporidia, invasion, polar tube, intracellular parasitism, germination
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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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Interactions between microsporidial parasites and the host cladoceran Daphnia pulex in a simple environment of a forest pond
Krylová, Pavla ; Petrusek, Adam (advisor) ; Hyliš, Miroslav (referee)
Among the most common endoparasites who infected small crustacean Daphnia pulex include microsporidia. These intracellular parasites appear to look like a simple single- celled organisms, but their cell structure and lifecycle prove the opposite. Microsporidia are species-specific. Although they infected most organisms of the animal kingdom, they are not yet sufficiently understood. This theses is inderectly followed up to the studies of waterflea Daphnia longispina and microsporidia Berwaldia schaefernai from the dam reservoirs The aim was to analyze closer microsporidian infection on host Daphnia pulex in a forest pool with simple enviroment, which included monitoring time dynamics of Daphnia population and identification infection caused by microsporidia Berwaldia singularis and yet unknown microsporidia labor-marked "HVH". Laboratory work included determination of zooplankton and parasites, calculation of prevalence, laboratory experiments with transmission of microsporidian infection between healthy and infected flea culture or by isolated spores. Genetic analysis of aquatic invertebrates from the forest pool, especially larvae of mite and mosquitos, helped make closer microsporidian life cycle and hypothesis about secondary hosts, for the presence of pathogen DNA using specific DNA...
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The parasitic microorganisms in immunodeficient and healthy population of honebees (Apis mellifra)
Bičianová, Martina ; Hubert, Jan (advisor) ; Titěra, Dalibor (referee)
Immunodeficient honey bee (Apis mellifera) colonies suffer from broad range of parasites including eukaryotic protozoa. Despite this fact, the eukaryotic parasites are still poorly documented in the Czech Republic. The presence of eukaryotic parasites (Nosema ceranae, Nosema apis, Crithidia mellificae and Apicystis bombi) was observed in different apiaries in the Czech Republic. The samples were taken in 9 apiaries in 53 beehives during the 2014/2015 season. From each beehive, 10 adult of honey bees were taken from the peripheral comb in triplicate. DNA was isolated from every sample of honey bees. The parasites were detected by polymerase chain reaction (PCR) with specific primers. The treatment fall of parasitic mite Varroa destructor was obtained from beekeepers for season of 2014. Crithidia mellificae was detected by 5 types of specific primers (SEF, SER; SSU, SSU rRNA, Cyt b, Tryp cyt b) and positive amplicons were cloned and sequenced. The obtained sequences were compared with GeneBank and showed similarity from 98-100% to sequences of Lotmaria passim (Trypanosomatid). Crithidia mellificae was not detected. L. passim had prevalence of 79,2% and is reported in the Czech Republic for the first time. Primer Tryp-cyt b is recommended for the routine detection of L. passim. Nosema ceranae was...
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Microparasites and fecundity of Daphnia at environmental gradients of canyon-shaped reservoirs
Hubová, Jana ; Petrusek, Adam (advisor) ; Macháček, Jiří (referee)
The original aim of my diploma thesis was examination of fixed samples of cladocerans from the Daphnia longispina species complex from reservoirs Vír and Vranov for the presence of 4 groups of microparasites: microsporidia, oomycetes, the protozoan Caullerya mesnili, and the yeast Metschnikowia bicuspidata. The next aim was to determine the effect of parasites on Daphnia fecundity, and determination of their spatial and temporal distribution within reservoirs. From the total number 4452 of examined Daphnia females, however, only 56 individuals were infected. This number was not sufficient for the planned analyses. As I recorded for all examined individuals the clutch size, I thus dealt to a large extent with an alternative issue: the temporal and spatial variation of, and the influence of environmental factors on Daphnia fecundity. Both studied reservoirs are characteristic by canyon-shaped profile that allows formation of environmental gradients on the horizontal as well as vertical axis. The results confirm that reservoir identity, season, and location within the reservoir (or gradient of food supply) have all significant effects on fecundity. During my work I have encountered difficulties associated with determining microparazites from fixed zooplankton samples. The appendix section of my thesis...
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Porcelain disease in crayfish: biology, impacts and distribution
Jelínková, Tereza ; Balcarová, Eva (advisor) ; Vávra, Jiří (referee)
The porcelain disease is a disease affecting crayfish caused by the microsporidium Thelohania contejeani. This intracellular parasite attacks muscle cells of the host and fills them with spores. This leads to white or opaque colouration of the abdominal muscle of crayfish, which is characteristic for this disease. The parasite gradually attacks more muscle fibers which probably leads to decrease of locomotor activity of affected crayfish and finally may result in their death. T. contejeani at low prevalences does not represent a real danger to crayfish populations and thus it is not as serious threat as crayfish plague. However, it has been associated with several cases of mass mortalities too. This thesis summarizes essential findings about infection by this parasite, analyzes its impacts to the host and factors that can influence transmission and progress of this disease. It also describes a life cycle of T. contejeani, summarizes records of the occurrence of this parasite and evaluates risks which it poses to crayfish populations. Finally, methods used to detect infection are described. Keywords: Thelohania contejeani, porcelain disease, crayfish, microsporidia, parasite, muscle
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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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