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Proteolytic systems of the blood fluke (Schistosoma mansoni).
Fajtová, Pavla ; Horn, Martin (advisor) ; Bařinka, Cyril (referee) ; Sojka, Daniel (referee)
Schistosomiasis is a serious parasitic disease caused by blood flukes of the genus Schistosoma. It is a global health problem with more than 200 million people infected and 750 million people at risk. Current therapy relies on a single drug, praziquantel, for which there are concerns of emerging drug resistance. Proteases of schistosoma are promising target molecules for the development of new therapeutic strategies against schistosomiasis. This work focuses on the comprehensive characterization of proteolytic systems of Schistosoma mansoni and determination of their role in the interaction with the human host. First, the major proteolytic activities secreted by individual developmental stages of schistosoma that parasitize the human body were classified using functional proteomics. This analysis demonstrated their complex and specific distribution with predominant serine and cysteine proteases and metalloproteases. Second, tegumental and digestive proteases, namely prolyl oligopeptidase and cathepsins B, C and D, were identified by chemical genomics as suitable target molecules for therapeutic intervention. Prolyl oligopeptidase was biochemically characterized using a recombinant protein, its effective inhibitors were developed as templates for antischistosomal drugs, and a biological role of the...
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Applications of CRISPR-Cas9 for genetic engineering in parasitic protists
Ročeň, Milan ; Tachezy, Jan (advisor) ; Rada, Petr (referee)
The CRISPR / Cas system serves as a cellular defense mechanism that protects bacteria and archaea from foreign DNA, especially bacteriophages. Its product forms a ribonucleoprotein complex, whose components are sgRNA and Cas endonucleases. Using sgRNAs, which are complementary to foreign DNA, this complex recognizes the DNA and Cas endonucleases induce double-strand breaks. This method is applied both in primary research, where it is applicable for functional analysis of proteins and the study of gene expression, as well as in applied research, where it can apply, for example, in the production of genetically modified organisms or genetically attenuated vaccines. This work summarizes the current knowledge of the CRISPR / Cas system and its application for gene manipulation in parasitic protists.
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Parasites And Their Preparation For School Utilization
Chlumová, Kateřina ; Říhová, Dagmar (advisor) ; Hanel, Lubomír (referee)
This diploma thesis deals with common parasites of game animals, animals and human occurring in the territory of the Czech Republic. It focuses on their fixation and general preparation of such specimens for usage in teaching of Science. The first part is devoted to the phenomenon of parasitism, and the history and definition of this concept. Systematic hierarchy of various unrelated parasitic groups is clarified. The second part provides the list of parasites, from the evolutionary original to the highly derived groups. We give an overview of their morphology and anatomy with the emphasis on their life cycle. The detailed knowledge of life cycle is essential for obtaining specimens. In each group, we indicate methods of collection and fixation in detail. The co called smartmicroscope represent the main point of the third part of proposed diploma thesis. Text of the thesis is supplemented with pictures taken by smart microscope, as well as by common microscope. KEY WORDS parasite, parasitism, mikropredatore, vector, fixation, preparation, smart mikroscope
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Střevní paraziti savců introdukovaných na Svalbard
BROŽ, Marek
The aim of this thesis was the research of parasites of mammals introduced to Svalbard: sibling voles, horses and dogs. Faeces of this three species were used for examination of intestinal parasites. Samples were collected in central part of Svalbard and examined in Czech Republic by microscopic and molecular methods for detection of protists and helminths.
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Výskyt endoparazitů u koní a možnosti prevence
ŠEJBOVÁ, Tereza
The aim of the diploma thesis is the analysis of the occurrence of endoparasites in horses and proposing prevention options using unconvential products and pasture management. In total 20 horses were included in the observation, who were all stabled on one farm. The observation took place during on pasture season. The first faecal examination was carried out for purpose of quantitative determination of parasites using the modified McMaster technique. The faecal analysis was carried out always before and after use of the homeopatihc and allopathic preparation. Horses were found to be infected by strongylides. Coprological analysis proved resistence to pyrantel embonate in 25 % of horses. After subsequent homeopathic treatment, the efficacy of this preparation was found in several horses, with EPG (eggs per gram) not exceeding 200. The effect of season on the rate of infection was monitored, when the effect of season wasn´t statiscially confirmed (p = 0,28). In assessing the effect of age on the intensity of infection, it was confirmed that older horses had lower parasitic infection rates than the young horses. After administration of active substance ivermectin to both groups of horses, a significant difference was observed when using this active substance against pyrantel embonate, which showed an excellent effect in 95 % horses. The homeopatic preparation of PVB etat vermieux can be used as an aditional preventive treatment during the year in horses that are sensitive to it. At the same time, it is important to regularly perform coprological faecal analysis and suitably choose an allopathic remedy, it is important to focus on preventive measures in horse breeding and pasture.
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Population genetics and speciation in Stylops (Strepsiptera)
Kodejš, Karel ; Straka, Jakub (advisor) ; Černá, Kateřina (referee)
This thesis is focused to population genetics of selected species of genus Stylops in Europe. Within this genus, five species was analysed by usage of two approaches - microsatellite analysis in Stylops ater and analysis of mitochondrial DNA in all five species. Mitochondrial gene for cytochome c oxidase (1st subunit) was used. For microsatellite analysis was performed bayesian clustering analysis and ABC approach (Aproximate Bayesian Computation). Mitochondrial markers were processed by making haplotype networks and demography analysis by computing Bayesian skyline plots. For Stylops ater, surprisingly low lewel of population subdivision was detected, yet with clearly differentiated population clusters from Scandinavia and baltic coast of Europe, which may imply period of isolation of these populations or relativelly recent population expansion and genetic differentiation due to lower population sizes. Next, hypotesis of possible temporal segregation of subpopulations of Stylops nevinsoni based on different aktivity period of their host species groups, was supported. In other three species, population subdivision was observed to be related either due to host specialisation (Stylops mellitae) or geographic consequences (S.nassonowi, S.spreta). Interesting finding is also detection of population...
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Secondary sexual ornaments and ectoparasites in the barn swallow (Hirundo rustica)
Wichová, Eliška ; Albrecht, Tomáš (advisor) ; Sedláček, Ondřej (referee)
Parasites could represent an important evolutionary driver and play an important role in a sexual selection. In the mate selection process, females use secondary sexual ornaments, which may reflect the parasite load and health condition of males. Females would benefit from choosing males with the most extravagant sexual characters, which indicate low levels of parasite infestation. A popular model species for sexual selection study is the barn swallow (Hirundo rustica). However there are no recent studies investigating the relationship between the level of the ectoparasitic infestation and the ornamentation of the barn swallow. Results of this thesis, based on analysis of ectoparasite load in 204 individuals show, that the level of infestation by feather mites is positively correlated with outermost tail feathers and the intensity of feather holes is negatively correlated with a breast coloration. The relationship between the abundance of ectoparasites and white tail spots was not found. The effect of ectoparasites on the survival of individuals or the nest initiation date was not observed. A possitive relationship between the individual seasonal change in feather mites infestation and brood size was detected. This implies a potential trade-off between the investments into parental care and defence...
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Fungal parasites as a factor affecting the dynamics of microalgal communities
Rosa, Jan ; Šťastný, Jan (advisor) ; Pichrtová, Martina (referee)
This bachelor thesis deals with fungal parasites which are an important factor that affects the dynamics of microalgal communities. Here I sum up the knowledge of parasites especially from Oomycota and Chytridiomycota along with a little focus on other groups. The basic characteristics of these fungal parasites, such as their host specificity and factors that affect their spreading ability between algal hosts, are summarized. The abiotic factors such as light and temperature, which are the most important factors causing epidemic, are mainly discussed. The fungal parasites can change the maximal population size and are able to alter the competitive relationships in microalgal communities. These factors are discussed in individual algal groups. The fungal parasites could be a negative economic player of the bioindustry and their future role in this industry is also discussed. Key words: parasite, host, host specificity, communities dynamics, algae, fungi, phytoplankton
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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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Parasitic protease SmCB2 as a target for the treatment of schistosomiasis
Bakardjieva, Marina ; Mareš, Michael (advisor) ; Mikeš, Libor (referee)
Blood flukes of the genus Schistosoma are parasitic trematodes that cause schistosomiasis, a serious disease afflicting more than 240 million people. The proteolytic system of schistosomes is essential for their viability: it participates in important processes during host-parasite interactions such as food digestion, invasion and tissue migration. Thus, schistosomal proteases are promising molecular targets for therapeutic intervention in schistosomiasis treatment. The thesis focuses on the protease cathepsin B2 from S. mansoni (SmCB2) which has not been studied in detail so far in terms of biochemical properties and biological function. Recombinant SmCB2 was prepared using yeast and bacterial expression systems and was chromatographically purified. Using an in vitro activity assay, the first effective inhibitors of SmCB2 were identified which inhibited its proteolytic activity in submicromolar concentrations. Specific polyclonal antibodies against SmCB2 were prepared and used for immunomicroscopic localization of this protease on the surface of the parasite. ELISA analysis demonstrated that SmCB2 is a parasite antigen recognized by the host immune system in the mouse model of schistosomiasis. The thesis provides valuable information about SmCB2 as a potential target molecule for synthetic...
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