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Influence of content of organic matter and calcium on immobilization of pharmaceuticals in soil
Kotradyová, Viktória ; Sedláček, Petr (referee) ; Klučáková, Martina (advisor)
This thesis aimed to highlight the problem of antibiotics and their presence in nature, particularly tetracycline. High global consumption, limited metabolism in the body and incomplete degradation in wastewater treatment plants represent the reasons why pharmaceuticals have been found in the environment. Their presence might have a negative impact on the natural ecosystem, including humans. Therefore, addressing their behaviour and finding solutions for their removal from nature seems important. The soil itself has a self-cleaning capacity, which may immobilise these pollutants into organic structures to prevent their subsequent migration. The presence of inorganic cations in soil plays an essential role in the sorption process of antibiotics. Calcium ions from the soil with whom the tetracycline forms chelate complexes, should be also taken into account. Regarding the practical part, tetracycline interactions with soil systems were studied in both sorption and desorption experiments. Four soils with different amounts of organic matter and calcium have been examined. Tetracycline was extensively adsorbed to all four soils. Desorption processes showed low efficiency and refer to stable adsorption onto soil. The experimental data were fitted by Freundlich and Langmuir isotherms, where the second mentioned is suitable for three of the four soils. Subsequently, sorption and desorption experiments were conducted in four different pH buffers (3, 5, 7 and 10). The effect of pH on sorption was not significant, however during desorption at lower pH higher desorbed levels of tetracycline were observed. Photodegradation of tetracycline in solution was investigated to determine the rate of degradation of tetracycline on light, where the results indicated that the concentration decreased by 22 % after 29 days compared to the applied concentration. Quantitative analysis was carried out using UV-VIS spectroscopy and liquid chromatography. In addition, the soil spectra of the FTIR were measured to obtain information on the sorption mechanism
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Biocrystallization as a new concept in development of piezoelectronics for use in biomaterials
Dobešová, Kateřina ; Plichta, Tomáš (referee) ; Sedláček, Petr (advisor)
This thesis focuses on biofilament methods and biocrystallization of bacterial cells producing polyhydroxyalkanoates, specifically the bacterium Cupriavidus necator H16. The main techniques used for cell orientation include spin-coating and acoustic waves. Biocrystallization was induced by temperature, osmotic, and acid stress. The degree of cell orientation was monitored by an atomic force microscope, and crystallization was determined by FTIR. Among other parameters, the thickness of the layers was evaluated using a profilometer. The amount of PHA in the bacterial cells was determined by gas chromatography. The preparation of crystalline ordered layers of biomaterials represents a route to piezoelectric biomaterials.
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Involvement of vibrational spectroscopy in the in-situ analysis of PHA in bacterial biomass.
Kevélyová, Barbora ; Samek, Ota (referee) ; Sedláček, Petr (advisor)
The aim of this thesis was the study of polyhydroxyalkanoates in bacterial biomass using the ATR-FTIR method. Furthermore, the PCA method was chosen to evaluate the measured data and observe correlations or differences among the spectra of various biomass samples. The bacterial producents were Aneurinibacillus thermoaerophilus H1, AH30 and AFn2, Cupriavidus malaysiensis DSM 25816, DSM 19416 and DSM 19379 and three strains of Schlegelella thermodepolymerans M 15344, DSM 15264 and LMG 21645. Various heteropolymers with different monomer content and total PHA content were investigated. The monomers in the samples were 3-hydroxybutyrate, 4-hydroxybutyrate, 3-hydroxyvalerate, 4-hydroxyvalerate, 5-hydroxyvalerate and 4-hydroxyhexanoate. As the reference method for accurate quantitative and qualitative determination of polyhydroxyalkanoates in the samples, gas chromatography was performed. The spectra obtained by the FTIR method were compared and significant peaks related to the qualitative properties of PHA, namely monomer composition and crystallinity, were searched for. The PCA statistical method was successfully applied to the collected data from FTIR, with the help of which the qualitative differences between the samples were monitored. The most significant differences were observed in the presence of monomers 4-hydroxybutyrate and 4-hydroxyhexanoate, and they were related to the crystallinity of the samples. The analysis did not show potential for semi-quantitative determination of PHA in biomass. Differences between microorganisms were not observed. The procedure for evaluating FTIR spectra using PCA could in the future be used in the study of biomass containing various monomeric units, especially hydroxyvalerates in case of their higher content within the sample.
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Utilization of advanced techniques of electron microscopy in altrastructural analysis of microbial cells and microbial biotechnology products
Havlíčková, Anna ; Sedláček, Petr (referee) ; Krzyžánek,, Vladislav (advisor)
In the theoretical part, this thesis deals with direct and indirect mechanisms of plant growth-promoting rhizobacteria, which is followed by a section on the selected model organism Azotobacter vinelandii. The second part of the theoretical section deals with electron microscopy starting with the possible interactions of the electron beam with the sample, through types of electron microscopes to various methods of preparing biological samples. The experimental part discusses the cultivation of selected bacterial cultures, the gelation of the cell cultures using chosen cross-linking agents (calcium chloride, malic acid, glucono--lactone with calcium carbonate) and, above all, various methods of preparation and analysis of pure and encapsulated cultures by the means of electron microscopy. In particular, cryogenic scanning electron microscopy and scanning transmission electron microscopy imaging were selected. Using both techniques, poly-3-hydroxybutyrate granules, which were confirmed to be elastic even at cryogenic temperatures, were observed inside the bacteria. Furthermore, it was found that cross-linking of extracellular alginate occurs even in samples without the addition of cross-linking agents. However, the hydrogel network present in these samples was significantly thinner than in samples with the addition of all cross-linking agents used. In this work, the density of the hydrogel network cross-linked with glucono--lactone was also compared. When using this agent with the addition of calcium carbonate, a significantly denser hydrogel network was observed in comparison with using the agent alone. Last but not least, when comparing the obtained results with the literature, it was found that the A. vinelandii bacteria form capsules, during the formation of the capsules the shape of the cells changes from an oval to a spherical bacterium and also the size of the given cell is reduced. From the resulting images, it was found that these capsules are made up of two layers, to which literature refers as electron-transparent intine and electron-dense exine.
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Utilization of novel strategies of alginate crosslinking in the development of next-generation bioinoculants
Gašparová, Dominika ; Pekař, Miloslav (referee) ; Sedláček, Petr (advisor)
Tato diplomová práce se zaměřuje na různé strategie síťování alginátu, s cílem posunout vpřed vývoj bioinokulantů nové generace. Zkoumání různých strategií gelování se aplikuje na pět komerčně dostupných alginátů a izolovaný bakteriální alginát. Experimenty zahrnující přípravu alginátového makrogelu sloužily ke stanovení výskytu a kvality síťování, které byly analyzovány amplitudovými testy. Výběr vhodných síťovacích činidel a stanovení jejich optimálních koncentrací byly klíčovými cíli. Síťovací činidla, která byla vybrána jako vhodná pro makro- i mikroenkapsulační procesy, byly ty obsahující chlorid zinečnatý, chlorid hlinitý, chlorid barnatý, síran vápenatý a chlorid železitý. Optimální podmínky síťování byly stanoveny při teplotě 7~°C a optimální délka doby síťování v rozmezí od 30 do 120 minut. Byly provedeny komparativní analýzy šesti vybraných síťovacích činidel a to prostřednictvím analýzy mechanických vlastností skrz squeeze test, termogravimetrickou analýzou, botnáním gelu a obrazovou analýzou. Síťování barnatým síťovacím činidlem prokázalo slibné výsledky při zachování obsahu vody během rehydratace, zatímco sítování zinečnatým síťovacím činidlem dosáhlo nejvyššího obsahu sušiny. Obrazová analýza odhalila menší gelové perly tvořené síťovacími činidly s obsahem hliníku a barya, což naznačuje jejich účinnost. Mikroenkapsulované gelové perly, zejména ty síťované s baryem, prokázaly potenciál díky jejich menší velikosti. Vrcholem pokusů bylo síťování bakteriální kultury, kde hliníková a barnatá síťovací činidla dosahovala nejmenších gelových perel. Obzvlášť síťování barnatým činidlem dosahovalo nejmenších velikostí s nejmenší odchylkou, indikujíc jeho efektivitu.
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Study on biodegradation of synthetic biobased polyesters
Guziurová, Pavlína ; Sedláček, Petr (referee) ; Obruča, Stanislav (advisor)
Tato práce se zabývá degradačními vlastnostmi polyesterových polymerů (PBS s inkorporovaným dilinoleovým sukcinátem v různých hmotnostních procentech) v simulovaných tělních tekutinách a odhaluje menší změny vlastností. Byla zkoumána také bakteriální degradace a analýza ukazuje významnější rozdíly ve výsledcích gelové permeační chromatografie (GPC) a skenovacího elektronového mikroskopu (SEM). Zatímco převažuje abiotická degradace, bakteriální prostředí vede k vyšším úrovním degradace než simulované tělní tekutiny, což naznačuje potenciální využití v obalové technice nebo zemědělství. Nižší náchylnost na abiotickou degradaci však otevírá nové možnosti pro další výzkum mechanismů bakteriální a enzymatické degradace pro ekologicky udržitelné alternativy v různých průmyslových odvětvích.
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Computer modeling of diffusional transport in hydrogel
Koláček, Jakub ; Sedláček, Petr (referee) ; Pekař, Miloslav (advisor)
This thesis focuses on the design of models applicable to the simulation of particle motion in a viscoelastic environment in COMSOL Multiphysics. Two approaches have been chosen for the design of these models: the first is the design of a geometry representing the porous structure of hydrogels and the second is the implementation of viscoelasticity using the mathematical concept of a continuous environment. Two elementary geometrical models are presented in this work – a three-dimensional periodic lattice and a spherical volume model. Furthermore, the possibilities of implementing the generalized Langevin equation in COMSOL Multiphysics are explored by using the interface for custom partial differential equations, by adding an auxiliary dependent variable, or by defining a custom external function written in the C language. The proposed geometric models did not prove to be suitable for the simulation of viscoelastic environment. An implementation using an external function seems to be the most promising, as it offers the most customization possibilities, and its implementation reflects the theoretical foundations. The thesis also includes a custom add-in written for COMSOL Multiphysics to facilitate the evaluation of simulation data and a modified Python script to calculate the complex shear modulus from MSD data.
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Material flow concept of a production line supply
Sedláček, Petr ; Dorušáková, Kamila (referee) ; Zeizinger, Lukáš (advisor)
The content of this thesis is a research analysis of individual concepts in the field of corporate logistics with an emphasis on intra-company transport. In the practical part, the current state of the production line loading is evaluated and new variants of line supply are made on the of the given parameters. The work also includes the creation of simulation models of all variants in the Plant Simulation program and conclusions are drawn based on the data obtained.
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Characterization of bacterial strains obtained in evolutionary engineering
Hrabalová, Vendula ; Sedláček, Petr (referee) ; Obruča, Stanislav (advisor)
This diploma thesis deals with application of evolutionary engineering on PHA producing bacterial strains. Two bacterial strains, Cupriavidus necator H16 and Halomonas halophila, were chosen for the evolutionary experiments. Copper cations (Cu2+) and sodium chloride (NaCl) were chosen as the selective pressure for C. necator H16; acetic acid (AA) and levulinic acid (LA) for Halomonas halophila. The adapted strains were during long-time evolutionary experiments characterized by GC-FID and SEC-MALS. The growth of the adapted strains was studied by the mean of optical density measurement. The amount of viable cells was determined by spectral FC after their expositon to selected stress factors. Specific enzyme activities of enzymes involved in citrate and glyoxalate cycle, enzymes generating NADPH, LA metabolism enzyme and PHA biosynthesis enzymes were determined. The adapted strains were compared with the wild-type of strains. The successfull adaptation of C. necator H16 adapted to Cu2+ was detected. Biomass and PHA production of both wild and adapted H. halophila strains cultivated in lignocellulosis waste were determined. It was found out that H. halophila adapted to the LA is capable of producing more PHA than the wild strain of this bacteria.
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Microrheology with Fluorescence Correlation Spectroscopy
Kábrtová, Petra ; Sedláček, Petr (referee) ; Mondek, Jakub (advisor)
A comparison of three passive microrheological techniques was made with the emphasis on FCS. Fluorescently labelled and unlabelled polystyrene particles were used to probe a microrheological response of glycerol solutions, Mili-Q water and sodium hyaluronate solutions. In addition, for FCS technique an approximated equation for calculation of MSD values was derived and verified. It was found that FCS outmatches current microrheological techniques of DLS and video-based particle tracking by its ability to gain a broader data range including the area in which, until now, it was impossible to describe a microrheological behaviour of samples reliably.
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