This diploma thesis deals with the change of production characteristics of model plants after application of vermicompost with addition of bacterial inoculum. In this work is elaborated a literature review focused on the process of vermicomposting, composting and stimulation of the root system of cultural crops using symbiotic or growth-promoting microorganisms. In addition, a practical experiment was carried out - an in-house vessel experiment with model plants was established, which was evaluated. The resulting data suggest that the most suitable variants in the formation of above-ground biomass were those containing vermicompost. The highest weight of aboveground biomass was achieved in vermicompost with the addition of Rizocore, where the production of aboveground biomass was increased 1.5 times compared to control soil.

Bacteria, the most dominant organisms on Earth, are an everyday presence in our lives. Symbiotic bacteria, which are present in the digestive tract of animals, usually have a beneficial effect on the body. On the opposite side of the spectrum are pathogenic species that cause more or less serious diseases around the world. In order to fight pathogens effectively, it is necessary to learn as much as possible about the molecular mechanisms by which bacteria respond to their environment, and also about the types of communication within bacterial populations that allow them to react to environmental changes as "multicellular" organisms. This Thesis consists of two main parts. In the first part, selected aspects of bacterial gene expression are characterized, using Bacillus subtilis and Mycobacterium smegmatis as model organisms. DNA-dependent RNA polymerase (RNAP) is the enzyme that is responsible for transcription of DNA into RNA, and thus plays a key role in gene expression. This Thesis deals with the structure of bacterial RNAP and important auxiliary factors (proteins and RNA) that associate with this enzyme and modulate its function. In the second part, the focus is on cell-to-cell communication, revealing which factors/mechanisms, including gene expression, affect this process in B. subtilis....

Respiratory system of the cystic fibrosis patients is affected by the defect in gene coding for protein transporter for chloride ions - CFTR ("Cystic fibrosis transmembrane conductance regulator"). The main complication of this disease is airways chronic inflammation, in particular caused by bacterium Pseudomonas aeruginosa. Due to asialylation of the lung surfaces the bacterial adhesion is facilitated, for example via lectin PAIIL. The ability of the chicken yolk antibodies to protect lung epithelial cells against Pseudomonas aeruginosa adhesion has been already proven. Therefore this thesis has mainly focused on the influence of the yolk antibodies specific against PAIIL on the development of infection in lungs of experimental animals. The objective was the optimization of the experimental model on which it would be possible to observe the infection development caused by luminescent bacteria strain in vivo using the optical tomography. At first the experiments have been performed on Wistar rats. Since the bacteria colonies in the rat lungs were not detectable in vivo on the available equipment, the rat experimental model showed up as not suitable. Further on only the mouse models were used. Experiments for the inhalation of the antibodies and intratracheal instillation of the bacteria suspension...

The main goal of this thesis was to evaluate the influence of additives on the qualitative indicators of grass silage. The evaluation concentrated in particular on lactic acid, pH and the degree of proteolysis. The selected qualitative indicators were monitored in 36 samples of grass silage from the region of South Bohemia. Silage samples were divided into three groups. The first group was a guide sample without any additives. The second group was treated with bacterial additives and the third group with bacteria-enzyme additives. Classification of silage samples was done according to "Norma 2004". Each sample was evaluated in terms of quality of fermentation and the content of nutrients. Statistically significant differences (p <0.05) were found in the content of lactic acid, pH and the degree of proteolysis between the group of silages without additives and the group treated with silage additives.

In the first part of my thesis I focus on mapping problems associated with antibiotic therapy and subsequent development of antibiotic resistance. Tracking resistance is based primarily on data collection and evaluation of the results set sensitivity from around the world. Antibiotic resistance is a natural phenomenon that can be observed in the evolution of microbes as one of the mechanisms of adaptation to new conditions in the environment. For this work I have chosen the following research questions. Do the incidence of ESBL strains in the České Budějovice Hospital a.s. increase over time? Are these values comparable to those achieved in another region, namely in Moravian hospitals the University Hospital of Olomouc, Ostrava University Hospital and Regional University Hospital of T. Bata in Zlin? The data collection I made in collaboration with the laboratory technicians and doctors at Hospital?s Bacteriology Laboratory in České Budějovice. Bacteries tested for the detection of ESBL production originated from biological materials, witch came from patients of hospital in České Budějovice. The first objective was to compare the results achieved in the České Budějovice Hospital in the period of 2007 to 2012. If we look at the total number of ESBL strains that have been isolated since 2007, values have upward trend. While in 2007 there were only 64 strains a year later, the number more than doubled. In 2010, the value soared to 281 tribes and in the year 2012, the number was 321 tribes. The incidence of ESBL strains in 2007 increased about five times. In the long term we can say the numbers have increasing tendency and the range of each species in the production of ESBL has significantly changed. In 2007, it was K. pneumoniae strains that dominated the statistics, but over time the strains of E. coli came forefront. Values of 2012 suggest that the presence of ESBL strains of K. pneumoniae is again almost equal to the number of E. coli strains. The second objective was to compare the results of the 2012 with study of the Prevalence of ESBL-positive Enterobacteriaceae in large Moravian hospitals. In the general overview of ESBL producers values in Hospital České Budějovice (5.23%) are comparable to those in Ostrava (4.9%) and in Zlín (4.3%). Number of strains in the Hospital in Olomouc (11.8%) is about twice as high as the numbers in České Budějovice. In this comparison the České Budějovice Hospital is one of the hospitals with a lower incidence of ESBL producers. The České Budějovice Hospital is below the national average, which originate from an elaborate system of care for patients with colonization or infection with ESBL strains, and from therapy control system using antibiotic center. These results may serve to the Hospital in České Budějovice for statistical purposes, and also for proposals for improving patient care. In the discussion, I pointed out the danger of the spread of resistant strains of bacteria in the community and also the associated risks that mentioned bacteria mean for patients injured in mass accidents or disasters. In these cases, number of infections including ESBL producers can penetrate through open wounds into the affected body. Unlike conventional sensitive bacteria those strains are resistant to commonly used antibiotics and thereby endanger the lives of people affected by the accident.

In the present study, the W sex-chromosome of the codling moth was studied by means of fluorescence in situ hybridization (FISH) with probes prepared from bacterial artificial chromosome (BAC), which were isolated from the codling moth BAC library. The BAC library was screened for clones derived from both the W and Z sex chromosomes using three sets of molecular markers of codling moth sex chromosomes. A total of 54 BAC clones have been obtained. In this work, only 3 W-derived BAC clones and 1 Z-derived BAC clone were further characterized by BAC-FISH mapping on chromosome preparations of pachytene oocytes; the other BAC clones have been retained for next studies. Whereas the Z-BAC probe provided a discrete hybridization signal on the Z chromosome, and surprisingly on the W chromosome, the W-BAC probes showed multiple hybridization signals distributed on the whole W chromosome, suggesting that they are mainly composed of repetitive sequences, which occur in multiple clusters on the W chromosome. The specific pattern of W-BAC hybridization signals along with the discrete signal of the Z-BAC enabled us to discriminate left/right orientation of both the W and Z chromosomes and examine specificity of W-Z pairing during meiotic prophase I.

Ľudské telo je prostredím pre život neuveriteľného množstva mikróbov. Niektoré z nich môžu spôsobovať rôzne choroby, ale ďalšie, napríklad črevný mikrobióm, sú pre život a zdravie človeka nepostrádateľné. Nanešťastie, črevný mikrobióm nie je detailne preštudovaný, pretože obsahuje tisíce rôznych druhov baktérií, z ktorých väčšina sa nedá kultivovať v laboratórnych podmienkach. Riešením tohto problému sú nové rýchle metódy sekvenovania v kombináciou s bioinformatickými nástrojmi na výpočet funkčného profilu baktérií vo vzorke. V tejto práci si predstavíme existujúce nástroje predpovedajúce funkčný profil, a následne navrhneme nový nástroj, ktorý môže implementovať konsenzus nad výsledkami existujúcich nástrojov, alebo sa môže jednať o úplne nový nástroj.

The aim of the work was testing of two types of magnetic mikrosheres functionalised with –COOH groups for the isolation of bacterial DNA. Isolation of DNA was carried out from crude lysates of cells prepared from pure culture of Lactobacillus paracassei RL-10 in the presence of binding buffer with 2 M NaCl and 16% PEG 6000. The influence of RNA degradation by enzyme RNase A on the amount of isolated DNA was investigated. It was estimated that RNA degradation affects the amount of DNA isolated. The amount of DNA depended on the type of microparticles. Higher amounts of DNA were isolated using particles with higher content of carboxyl groups. DNA applicable in PCR was isolated using both types of microsheres. In next part of the work, microparticles functionalised with –NH2 groups were used to DNA isolation using electrostatic forces. It was shown that buffer with lower pH is suitable for DNA adsorption onto magnetic microparticles.

Recently much attention is given to lanthanides and their complexes as excellent catalysts for cleavage of nucleic acids. The thesis has been focused on the cleavage of plasmid and bacterial DNA by ions Nd3+ and Y3+ and by different carriers containing the lanthanide compounds. The creation of single-stranded nicks and double-stranded ones in the plasmid DNA molecules was studied by agarose gel electrophoresis. Verification of the cleavage of bacterial DNA was made by polymerase chain reaction using primers specific for the domain Bacteria and genus and species-specific primers. The results will be used in the development of the method that will allow perfect carriers`s coverage verification with the magnetic perovskit nucleus and other carriers with the lanthanide compounds.

This diploma thesis deals with testing, optimazing and comparing methods for the identification of bacteria producing polyhydroxyalkanoates. Work included cultivation and microscopy methods, wherein the bacterial cells were stained with lipophilic dyes Nile red and Sudan black. Further, we also used flow cytometry and spectroscopic methods - Raman spectroscopy and infrared spectroscopy with Fourier transformation, and molecular biological methods, which analyzed the presence of a gene encoding PHA synthase (phaC) by polymerase chain reaction (PCR). PCR assay consist of two reactions, the firt on eis based on amplification of phaC gene along with 16S rRNDA gene, which is common for all the bacteria (multiplex PCR). The second reaction is focused on specific amplification of PHA synthase catalyzing biosynthesis of mcl-PHA. In order to overcome false positive results typical for methods analyzing genotype and also to avoid false negative results occuring in fenotype analyzing methods, the best strategy is to combine both aproaches. According to our results, analysis of presence of phaC gene by PCR can be combined with methods capable of determining presence of PHA in bacterial cells. For this purpose, Raman microspectroscopy seems to be very promising tool, since it is able to detect low content of PHA in cells and PHA can not be confused with other lipid metabolites. The results provide an overview of test methods, their advantages and disadvantages and also to compare different criteria according to which it is possible to choose the method of identification in depending on the adjustable requirements.