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Use of microscopy and flow cytometry to molecular characterization of carotenogenic yeasts.
Vacková, Hana ; Skoumalová, Petra (referee) ; Márová, Ivana (advisor)
The aim of presented bachelor thesis is to optimize the methods for study of yeasts using flow cytometry and fluorescence microscopy. The theoretical part deals with the structure of yeast cells, principles and application of flow cytometry and fluorescent microscopy. Use of microscopy and flow cytometry in study of pigmented microorganismswas reviewed too. In the experimental part methods optimization was performed on yeasts Cystofilibasidium capitatum, Rhodotorula glutinis, Sporobolomyce sroseus, Sporobolomyces shibatanus. Further, the growth curve and the viability of yeasts under the influence of stress factors (freezing and the heat shock 60 C) was measured. Cystofilobasidium capitatum has shown to be more resistant to stress than Rhodotorula glutinis. It was found that there is the possibility of using methylene blue instead of propidium iodide in yeast cell analysis by flow cytometry.
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Úloha Adenylát kinázy 1 v aktivaci a metabolismu imunitních buněk larev \kur{Drosophila melanogaster}
KAISLEROVÁ, Nikola
The aim of this thesis was to study the role of Adenylate kinase 1 (Ak1) in the immune system of Drosophila melanogaster larvae upon the infection by parasitoid wasp Leptopilina boulardi. Using the immune specific Ak1 RNA interference, it was analyzed the effect of Ak1 reduction on the immune response and viability of Drosophila. The importance of Ak1 was also evaluated within the metabolism of immune cells. It has been shown that Ak1 is crucial in energy metabolism of immune cells and important for the proper functioning of immune system.
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Ecophysiology of microscopic fungus Pseudogymnoascus destructans
Homutová, Karolína ; Kubátová, Alena (advisor) ; Baldrian, Petr (referee)
A microscopic fungus Pseudogymnoascus destructans (Ascomycota: Pseudeurotiaceae) causes illness known as white-nose syndrome (WNS) causing death of bats during hibernation. The illness occurs in the North America and in Europe. The fungus is characteristic by asymmetrically curved conidia, by slow growth and growth at low temperatures (below 20 řC). The aim of this study is to clarify properties responsible for unique ecelogy of Pseudogymnoascus destructans by comparison with ecological related or unrelated pathogenic or nonpathogenic fungi. This part includes study of tolerance to physiological stresses and recognition of spectrum of utilizating nutrients (compounds of carbon, nitrogen, phosphorus, sulphur and nutrient supplements). Testing to physiological stresses should help to estimate a potentiality of fungus to spread out of caves. The last aim is to develop a selective isolation medium for P. destructans. Influence of several types of physiological stress (e.g. UVA, UVA with UVB, 25 řC, 30 řC, 37 řC and dryness) was investigated with fluorescent stain propidium iodide (PI) by flow cytometry. The spores of Pseudogymnoascus destructans and three fungi from underground spaces were not viable after 3 weeks at 37 řC. Other stresses did not cause a decreasing of viability or some stresses caused...
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Faktory ovlivňující životaschopnost a růst telat masného skotu
Hlaváčková, Marta
The literature review describes factors affecting the viability and growth of beef cattle calves. The aim of the literature review was to summarize information of various authors and to elaborate an overview of factors such as maternal nutrition, its health status, the course of calving or nutrition of cattle calves. The thesis also contains my personal practical information and experience in the care of little vital calves that I have gained during my life on the farm. The practical part considers the analysis based on the causes of the reduced viability of calves in our cattle farming. By measuring and weighing I evaluated the influence of various factors on growth ability. Three calves with reduced viability are compared (healthy calf of sick mother, sick calf of healthy mother, sick calf of sick mother) with a control calf (healthy calf of healthy mother). In the final part of the thesis the results obtained are appraised.
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In vitro cultivation of the trematode species Trichobilharzia regenti
Vrbová, Kristýna ; Kašný, Martin (advisor) ; Skelly, Patrick (referee)
The class Trematoda includes many pathogenic representatives. Main subject of this thesis, avian schistosome Trichobilharzia regenti, is a close relative to the important human pathogen Schistosoma mansoni (family Schistosomatidae). In vitro cultivation of trematodes enables closer understanding of their biology and parasite- host interactions; however, no trematode species has been successfully kept in vitro from the egg stage to the adults producing eggs. Many studies are focused on the problematic of S. mansoni cultivation, but data concerning T. regenti cultivation remain scarce. Only the ability of T. regenti cercariae to transform into schistosomula in vitro was documented, with following survival in a culture medium for a few days. Comparison of eight transformation methods was performed with T. regenti cercariae. Based on the number of tailless cercarial bodies obtained, five transformation methods were selected for further evaluation of the early schistosomula characteristics (glycocalyx shedding, penetration glands emptying and survival in vitro). It was observed that the largest quantity of cercarial bodies can be obtained by using a syringe needle or the BeadBeater cell disrupter. The largest quantity of schistosomula meeting the criteria of early schistosomulum was recorded after...
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Ecophysiology of microscopic fungus Pseudogymnoascus destructans
Homutová, Karolína ; Kubátová, Alena (advisor) ; Baldrian, Petr (referee)
A microscopic fungus Pseudogymnoascus destructans (Ascomycota: Pseudeurotiaceae) causes illness known as white-nose syndrome (WNS) causing death of bats during hibernation. The illness occurs in the North America and in Europe. The fungus is characteristic by asymmetrically curved conidia, by slow growth and growth at low temperatures (below 20 řC). The aim of this study is to clarify properties responsible for unique ecelogy of Pseudogymnoascus destructans by comparison with ecological related or unrelated pathogenic or nonpathogenic fungi. This part includes study of tolerance to physiological stresses and recognition of spectrum of utilizating nutrients (compounds of carbon, nitrogen, phosphorus, sulphur and nutrient supplements). Testing to physiological stresses should help to estimate a potentiality of fungus to spread out of caves. The last aim is to develop a selective isolation medium for P. destructans. Influence of several types of physiological stress (e.g. UVA, UVA with UVB, 25 řC, 30 řC, 37 řC and dryness) was investigated with fluorescent stain propidium iodide (PI) by flow cytometry. The spores of Pseudogymnoascus destructans and three fungi from underground spaces were not viable after 3 weeks at 37 řC. Other stresses did not cause a decreasing of viability or some stresses caused...
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The business plan for the establishment of limited liability company
Kubátová, Barbora ; Trachta, Václav (advisor) ; Dvořák, Jiří (referee)
The aim of this final written exam is to create the business plan of setting up limited company. Knowledge of economy, financial management and marketing are used for detailed research. This thesis is divided into two parts. The first part contents theoretical knowledge which are connected with the business target and there is also made concrete business plan. The second part is practice and there is concrete business plan which has been put together on the foundation from the previous research. The aim of this thesis was to found out the ability of survive and profitability of the limited company.
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Factors influencing seed germination, emergence and mortality in soil seed bank of Bromus sterilis L.
Žďárková, Veronika ; Soukup, Josef (advisor)
Bromus sterilis L. (barren brome) spreads rapidly in many European regions. In the Czech Republic, its importance has increased dramatically over the past 10 years. Barren brome is reported as a problem weed in winter crops such as winter wheat, winter barley and oil seed rape, in vineyards and in other cultivated places. Barren brome has been becoming troublesome weed of winter cereals mainly in reduced soil tillage systems. The factors, that are important for its spreading and adaptability under different environmental conditions, are dormancy and germination. Optimal timing for seed germination varies with respect to natural conditions and it is determining for plant development. These traits are adapted to different conditions and habitats; therefore the dormancy and germination response patterns to those conditions vary significantly. Recently there are not many information available on germination behaviour, therefore this study was focused on seeds of barren brome and its characteristics, which were collected in different regions of the Czech Republic. Seeds were investigated under different temperatures, light regimes and water stress in a wide range of conditions. The following characteristics of seeds were studied the dormancy, the dynamics of germination, the temperature optimum, the age of seeds and dynamics of emergence from different depths and persistence in the soil profile under field conditions. Dormancy and germination are influenced by external conditions. The dormancy of seeds barren brome is very short or missing. The germination is influenced by light; the seeds germinated better under darkness than under light regime. The primary dormancy of B. sterilis was short and the seeds needed only three weeks for after-ripening. The seeds of B. sterilis showed broad ecological valence to hydrothermal factors germinating in the wide range of 3 to 35 °C. The temperature optimum is 5-23°C. The germination was only slightly influenced in an environment with low water potential. Germination was limited under water stress only at lower temperatures under 10°C. The response to light at various temperatures showed that seeds germinated better in darkness in all temperatures regimes, than in alternating light regime, especially at lower temperatures. The emergence declined significantly with burial depth (under 40 mm). The seeds were not able to survive in the soil seed bank for a longer time and fall seeds lost viability after 1 year burial in soil profile. These results may be of value for development of predictive models and understanding period when weed control may be most feasible.
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Methods of storage of honeybee semen: evaluation of honeybee spermatozoa viability with flow cytometry.
Savvulidi, Filipp ; Titěra, Dalibor (advisor) ; Jan, Jan (referee)
Assessment of semen quality (sperm viability) is an essential for the efficient honeybee semen storage. The motility of spermatozoa is not the ultimate parameter of sperm viability. Instead, the viability of spermatozoa is assessed by testing their structural or functional plasma membrane integrity. These assays are based primarily on the technique of microscopy evaluation of either fluorescent DNA dyes or hypo-osmotic "tail-coiling" reaction of spermatozoa. However, the technique of microscopy evaluation is time-consuming, subjective and often is not accurate. On the other hand, the technique of flow cytometry is rapid, objective and precise. Nowadays, this technique is affordable for many institutions, as the price of "benchtop" flow cytometer is relatively low. To best of our knowledge, there is currently no flow cytometry assay available for evaluation of functional integrity of honeybee spermatozoa. Here we report the establishing of novel flow cytometry assay for simultaneous evaluation of structural and functional plasma membrane integrity. DRAQ7, the new fluorescent DNA dye was used to test the structural integrity of plasma membranes. The hypo-osmotic "tail-coiling" reaction was used for flow cytometric assessment of functional integrity of spermatozoa plasma membranes. The combination of both is a valuable tool for rapid and precise evaluation of sperm viability in honey bees.
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Vyhodnocení vlivu příbuzenské plemenitby na životaschopnost a dlouhověkost koní Převalského
Nogová, Barbara
The literary overview in the first chapter focuses on the occurrence and discovery of the Przewalski horse. It also deals with their own characteristic. Information about the lifespan of Przewalski horses and individual breeding lines are specified too. Attention was also engaged to the topic of inbreeding and inbreeding depression. The last chapter of a literary overview discusses the current state of the breed. The aim of these theses was to process the data from the general studbook of the Przewalski horses and to evaluate the effect of inbreeding on the length of life of these horses. Data required to statistical evaluation was obtained from the International studbook of Przewalski horse from 2008 and from publicly available internet version of these book. The database contained informations about a sex, date of birth, date of death, lifespan, and the value of the coefficient of inbreeding. Thus obtained data were processed using the program Microsoft Excel 2010 and statistically evaluated in the programs of the UNISTAT 6.5 and SAS 9.1.4. Used to be a basic statistics functions, the Tukey-B test, generalized linear model and Pearson's correlation. Assessing the results of the statistics was detected highly significant effect of sex, year of birth, and the values of the coefficient of inbreeding on the lifespan of Przewalski horses. However, for foals of Przewalski horses, that lived more than two years, has been demonstrated only effect of sex on the lifespan.
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