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Bacterial biofilm formation
Sochorová, Zuzana ; Weiser, Jaroslav (advisor) ; Zikánová, Blanka (referee)
Biofilms are formed by microorganisms living together in a hydrated extracellular matrix. Formation of such clusters of bacteria brings many benefits. The increased resistance to antibiotics is the main one. Creating a biofilm is analogous to the development of multicellular organisms. Biofilm cells communicate with each other with signaling molecules. Signaling molecules make the biofilm more compact structure. Extracellular matrix, in which bacteria live, maintains biofilm structure, affects cell adhesion and protects cells against environmental influences. Bacteria also interact with the environment through the extracellular matrix. The matrix is composed of various biopolymers and proteins. Biofilms are a common cause of infections associated with implants. There are several ways to prevent bacteria in biofilm to antibiotics. These include a slow diffusion of substances, a slow growth or an adaptation to stress. The formation of persistent cells that are tolerant to the antibiotics is the cause of their survival as well as a new population may arise from them. The surface proteins are important elements for the formation of biofilms, they facilitate adhesion and subsequent establishment of biofilm. A protein Bap was the first identified protein. Later its homologues were found, such as BapA,...
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Carbapenemases and their molecular genetic detection in bacteria causing severe hospital acquired infections
Vavrová, Jolana ; Dřevínek, Pavel (advisor) ; Zikánová, Blanka (referee)
The aim of this study is to characterize β-lactamases that hydrolyze carbapenems. They belong to molecular classes A, B and D and their new variations are still being identified. The production of enzymes hydrolyzing β-lactam antibiotics is common among Gram-negative bacteria which include also serious nosocomial pathogens such as Pseudomonas aeruginosa, Acinetobacter baumannii or members of Enterobacteriaceae family. The importance of the carbapenemases in clinical practice underlines the fact that carbapenems are broad-spectrum antibiotics, which are considered the reserve antibiotics. An increase in the number of cases of the carbapenem resistance is caused mainly by spread of genes encoding carbapenemases. Detection of carbapenemases and surveillance of their occurrence are both essential not only for optimal management of patients' care, but also for control of the spread of bacterial strains producing carbapenemases.
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Fungi associated with decomposing wood in temperate forests
Štercová, Lucie ; Baldrian, Petr (advisor) ; Zikánová, Blanka (referee)
Wood decaying fungi are an essential part of all forest ecosystems. On their functioning depends a number of other organisms which use substances produced during decay of wood. Enzymes that degrade wood represent indispensable tool for fungi in converting structural compounds of wood to water and carbon dioxide. That makes them an essential part of the carbon cycle in nature. The dynamic of fungal communities on decaying wood is determined by a range of abiotic and biotic factors. The variability of microclimatic conditions, differences in the wood humidity and a gaseous mode are identifying stress factors which define the presence of species adapted to those conditions. An important factor, with which we can partially predict community composition, is the degree of decomposition of wood and the history of a species on the substrate. Another indicator, which may partly explain the composition of the community is a way of dying of a tree and a type of decaying substrate. Often, the development of a community follows from the primary colonizers, with high tolerance to unfavorable conditions, through the secondary colonizers, which have the ability to obtain the substrate over primary colonizers, but require more stable microclimatic conditions, to late colonizers, who are adapted to stress factors as...
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Lessons from nature - preparation of hybrid bioactive compounds
Vobruba, Šimon ; Janata, Jiří (advisor) ; Zikánová, Blanka (referee)
Secondary metabolites are biologically active compounds produced mainly by microorganisms. They are not essential for survival of producing strains, however, they significantly affect their physiology and ecology. They are frequently used in pharmacology, biology and chemistry. The present work describes the current state of knowledge concerning origin and evolution of secondary metabolites. The secondary metabolites biosynthetic genes are usually organised in clusters. The basic mechanisms of secondary metabolite gene clusters modification are gene mutations or intragenic rearrangements. These mechanisms are typically involved in natural evolution of gene clusters coding for secondary metabolites with modular type of biosynthesis. The subclusters of different origin can also fuse to form a new hybrid compound biosynthetic gene cluster. Similar evolutionary event probably occurred also in case of biosynthesis of two model groups of natural compounds - lincosamides and pyrrolobenzodiazepines. Analogous approaches are used in genetic engineering to construct producers of new more efficient bioactive compounds. Examples of such genetic modifications of gene clusters involved in the biosynthesis of compounds from nonribosomal peptides, polyketides and lincosamides groups are described. Possible future...
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Microbial community in sediments of a stream contaminated by pharmaceuticals
Brťková, Hana ; Cajthaml, Tomáš (advisor) ; Zikánová, Blanka (referee)
Pharmaceuticals are micropollutants, that enter the environment mainly through Wastewater Treatment Plants (WWTPs). In this work microbial community has been studied in sediments of a stream, which is located near a WWTP. This sediment is contaminated with pharmaceuticals. The subject of this thesis was to determine the presence of pharmaceuticals and microbial community in this study site and to point out possible relationships between these factors. Twelve pharmaceuticals were identified at concentrations reaching levels of ng/g. The concentrations of the compounds form a gradient that decreases with the distance from WWTP. Microbial biomass was estimated using the analysis of phospholipid fatty acids and microbial community was described using next-generation DNA sequencing. The analysis of phospholipid fatty acids pointed out, that with the increasing distance from WWTP the amount of microbial biomass decreases. DNA sequencing revealed large microbial diversity in the studied sediment. For evaluation of the relationship between the microbial community and pharmaceuticals in the stream sediment, Principal Component Analysis (PCA) was used. The result of PCA showed, that in the stream sediment (depth 10-30 cm), Betaproteobacteria negatively correlated with triclosan and Clostridia negatively...
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Colonization of human lung by non-pathogenic streptomycetes
Herbrík, Andrej ; Petříčková, Kateřina (advisor) ; Zikánová, Blanka (referee)
Streptomycetes, primary soil saprophytic microorganisms are at the center of interest in many research groups, mainly because of their ability to produce a wide range of biologically active substances useful in medicine, biotechnology and agriculture. The marginal, and little explored areas are the interactions that streptomycetes create with humans. Recent metagenomic studies have shown that streptomycetes colonize the skin, the respiratory and possibly the urogenital tract of humans. In addition to apparent pathogens such as S. somaliensis and S. sudanensis, the clinical impact of these streptomycetes on human health is unknown. For this reason, a unique collection of non-pathogenic streptomycetes isolated from human clinical specimens was developed. The isolates were collected by the National Reference Laboratory for Pathogenic Actinomycetes in Trutnov, Czech Republic. On the basis of pilot studies, an isolate labeled TR42 was selected from the Trutnov collection, showing a very broad spectrum of biological activities. This strain was isolated from sputum from a patient with unknown respiratory diagnosis. The TR42 strain exhibits considerable biotechnological potential and after following a thorough study, it could be a source of new biologically active substances with pharmaceutically...
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Antibacterial and antiadhesive properties of carbon nanomaterials
Budil, Jakub ; Lišková, Petra (advisor) ; Zikánová, Blanka (referee)
Increasing interest in industrial and medical applications of carbon nanomaterial leads to the need to examine its interactions with living systems. Nanocrystalline diamond (NCD) films possess high mechanical and chemical stability which, together with its biocompatibility with human cells, enables applications in human body. Some of carbon nanoparticles possess strong antibacterial activity. In this work the effects of NCD with hydrogen, oxygen and fluorine termination deposited on glass and silicone on adhesion of gram-negative bacteria Escherichia coli K-12 in mineral medium is described and the impact of cultivation medium on effects of NCD films is compared. Prior the growth of the E. coli biofilm on NCD films, the method for quantification of biofilm using crystal violet staining and the method for biofilm cultivation in mineral medium were optimised. The properties of NCD film are independent on the base substrate. Hydrogen and fluorine terminated NCD films show antiadhesive properties only in mineral medium but not in complex medium. This is explained by formation of a conditioning film on the surface of the NCD film during cultivation in complex medium. On the other hand, O-NCD film supports bacterial adhesion in both cultivation media. Second part of this thesis is dedicated to carbon...
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Role of autophagy in yeast cell adaptation
Brádlerová, Michaela ; Kuthan, Martin (advisor) ; Zikánová, Blanka (referee)
Autophagy is an evolutionarily conserved degradative pathway. Autophagy occurs constitutively at a basal level and it is involved in the recycling and turnover of damaged or superfluous organelles and proteins. It has a critical role in cellular homeostasis. Autophagy can be induced in response to starvation or other types of stress. Induction of autophagy during these conditions has a major role in protection and adaptation of the cell. Autophagy needs to be properly regulated. A wide range of diseases is associated with dysregulation of autophagy. Better understanding of autophagy mechanisms can help to develop strategies designed to modulate autophagic responses occuring in a number of diseases. This work is focused on current knowledge of main types of autophagy and how autophagy helps yeast cells to adapt. Key words: autophagy, yeast, degradative pathway, adaptation, TORC1
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Specific features of streptomycetes isolated from human tissues
Kodatová, Anežka ; Petříčková, Kateřina (advisor) ; Zikánová, Blanka (referee)
This bachelor thesis focuses on activities of the bacterial genus Streptomyces in relations to plants, animals and human. Many cases of their symbiotic relationships with plants are known, but there is also a common plant pathogen S. scabies. As a primary soil bacteria, their association with animals is infrequent, but not exceptional. Symbiotic relationships have been observed for example in several insect species. Streptomyces have been repeatedly isolated from the human microbiome, however only a little information about their influence on human body is available. Due to the wide production of secondary metabolites affecting other microorganisms, their role can be assumed as significant. The relationship between human Streptomyces and human could have a parallel in the Streptomyces symbiosis with plants and insects, but it can also have certain effects on pathogenesis. The ability of beta haemolysis was demonstrated in tested strains. In addition, there are cases of serious diseases caused by Streptomyces. Next to the relatively common actinomycosis in the regions of Sudan and India, diseases such as pneumonia or abscess were reported.
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