This diploma thesis deals with selected thermophilic representatives of genera Geobacillus, Saccharococcus and Bacillus, taking screening of its biotechnological potential into account. Bacteria from the first two genera came from Czech and German collection of microorganisms, while bacteria of genus Bacillus were natural isolates. Researched strains were examined from a viewpoint of carbon source utilization and furthermore, production of biosurfactants, extracellular hydrolytic enzymes (protease, amylase, lipase, cellulase, xylanase), organic acids, antimicrobial agents and microbial plastics – polyhydroxyalkanoates was also tested. Bacteria S. thermophilus, G. uzenensis and G. zalihae evinced a substantial ability of biosurfactant production. Strains G. jurassicus, G. uzenensis, G. gargensis and G. lituanicus were capable of intensive production of all tested, technologically significant enzymes. Highest antimicrobial effects were reached with bacteria G. stearothermophilus and G. thermocatenulatus. Largest production of acetic acid was achieved with G. jurassicus and lactic acid with G. thermodenitrificans. Ability to produce polyhydroxyalkanoates was proved at genotype level by some cultures only, however at fenotype level, response was negative. On the contrary, bacteria genus Bacillus were able to produce polyhydroxyalkanoates, although in small amounts under given circumstances. With remaining researched metabolites, production ability was considerably lower, compared to genera Geobacillus and Saccharococcus.

This thesis deals with the study of polyhydroxyalkanoate biosynthesis as a microbial product with the potential to replace current conventional plastics made from petroleum. The dissertation thesis is elaborated in the form of a discussed set of already published publications, which are then part of the thesis in the form of appendices. The work builds on relatively extensive knowledge in the field of polyhydroxyalkanoate production and brings new facts and possible strategies. Various possibilities of analysis of polyhydroxyalkanoates using modern methods were tested in this work, which brings especially speed, which can be crucial in real-time evaluation of production biotechnological process. Raman spectroscopy has proven to be a very promising technique for rapid quantification of PHA. Furthermore, the work deals with valorisation of waste of food and agricultural origin. Emphasis is placed on methods of detoxification of lignocellulose hydrolysates. In this context, adsorption of inhibitors to lignin was first used as an alternative to other detoxification techniques. Due to detoxification, selected production strains Burkholederia cepacia and B. sacchari were able to utilize softwood hydrolyzate for PHA production. In the next part of the work was also tested the possibility of using chicken feathers as a complex source of nitrogen. Evolutionary engineering was also used as a possible strategy to eliminate the inhibitory effect of levulic acid as a microbial inhibitor that results from the hydrolysis of lignocellulosic materials. Adaptation experiments were used to develop strains exhibiting higher resistance to levulic acid and the ability to accumulate a higher 3HV copolymer from the original wild-type C. necator strain. Another promising approach tested in the work was the use of extremophilic microbial strain, which leads to a reduction in the cost of biotechnological production. Selected Halomonas species have shown high potential as halophilic PHA producers. The final part of the thesis was devoted to the selection of the production strain with regard to the properties of the resulting PHA. The Cupriavidus malaysiensis strain was selected to produce a P(3HB-co-3HV-co-4HB) terpolymer which revealed significant differences in material properties over P3HB.

This thesis consists of one chapter accepted for publication in a book and four papers published in international journals with impact factors. All of the contributions deal with the role of wood ants in energy and nutrient fluxes in forest ecosystems. Wood ant nests are known as hot spots of carbon dioxide (CO2) production and are also thought to affect methane (CH4) flux. Stable high temperatures are maintained in ant nests even in cold environments. This study is focused on quantification of CO2 and CH4 flux in wood ant nests, contribution of ants and microbes to CO2 production, properties of nest material that affect CO2 production and the role of ants and microbes in the maintenance of nest temperature. The research was conducted in temperate and boreal forests inhabited by wood ants (Formica s. str.). Gas fluxes were measured either by an infrared gas analyser or a static chamber technique. Ants and nest materials were also incubated in a laboratory. Material properties potentially influencing CO2 flux, such as moisture, nutrient content or temperature were determined. According to the results, CH4 oxidation was lower in wood ant nests than in the surrounding forest soil suggesting that some characteristics of ant nests hinder CH4 oxidation or promote CH4 production. Wood ant nests clearly are hot...

The thesis deals with the use of dissolved air flotation (DAF) in water treatment technology in the Czech Republic. It summarizes the knowledge gained at pilot plant studies as well as the experience with full-scale flotation units which have been built in potable water treatment plants in the Czech Republic in years 2006-2018. This thesis provides a broad background of experimental studies of a modern separation process, which does have great advantages in the removal of natural organic matter and microorganisms from water in the drinking water treatment industry. In this thesis, there are presented some examples of the excellent efficiency of the microorganisms removal, both under the experimental conditions as well as water treatment plants, which were improved by DAF units installation in full scale. In addition to the high separation efficiency, it is shown, how DAF units could influence the subsequent separation steps as for example filtration through the granular media. The negative influence of the pre-ozonization on the DAF separation efficiency in Hradec Králové WTP is demonstrated too. The results of the pilot experiments support the idea, that the key condition for high removal efficiency is the optimal coagulation process chemistry. Design of pilot experiments was based on the factorial planning theory and some results are discussed in this thesis. The very big portion of the pilot experiments was made for estimating of the full-scale DAF design parameters before the water treatment plants reconstruction phase

Charles University Faculty of Pharmacy in Hradec Králové Study program: Healthcare Bioanalytics Author: Bc. Adéla Diepoltová Supervisor: RNDr. Klára Konečná, Ph.D. Title of thesis: Optimization of the method for sensitivity evaluation of biofilm- forming staphylococci against candidate antimicrobial compounds Background: The aim of this thesis was to optimize approach for in vitro formation of staphylococcal biofilms on the pegs and on the wells of the 96-well panel as an analogous approach to commercially available Calgary Biofilm Device system. The aim of the Experiment 1 was to evaluate incubation conditions (such as impact of a growth medium, incubation mode, optical density of the starting bacterial inoculum and type of surface) leading to maximal biofilm formation of two biofilm producer strains with unknown biofilm phenotype and one staphylococcal strain known as strong biofilm producer. The most advisable conditions were used in incubation process of Experiment 2. This work should propose the approach leading to in vitro formation of the most voluminous staphylococcal biofilms exploitable for candidate drug antimicrobial activity testing. Methods: Spectrophotometric measurement of the crystal violet colour extracted from wells with fixed and stained Staphylococci to evaluate the ability to...

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