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Metodika odběru a izolace bakterií, kvasinek a plísní z fotografických materiálů
Demnerová, Kateřina ; Kadavá, Jana ; Purkrtová, Sabina ; Savická, Dana ; Sýkorová, Hana ; Ďurovič, Michal ; Benetková, Barbora ; Hricková, Kateřina ; Koukalová, Lucie ; Nováková, Martina
Metodika odběru a izolace bakterií, kvasinek a plísní z fotografických materiálů navrhuje optimální odběrový materiál a techniku odběru vzorku pro mikrobiologickou analýzu fotografických materiálů, která by v první řadě nepoškodila jejich povrch citlivý především na mechanické poškození, a zároveň vykazuje dostatečný záchyt mikroorganismů z povrchu. Je určena pro konzervátory, restaurátory a výzkumné pracovníky, kteří se věnují preventivní péči o fotografické sbírky v archivech, knihovnách, muzeích a galeriích.
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Soil microbial communities in agroecosystems and natural habitats contributing to resistance and resilience of the soil environment
Sarikhani, Ensyeh ; Kopecký, Jan (advisor) ; Demnerová, Kateřina (referee) ; Větrovský, Tomáš (referee)
Ensyeh Sarikhani Soil microbial communities in agroecosystems and natural habitats contributing to resistance and resilience of the soil environment. Summary The control of common scab of potatoes (CS) includes resistant varieties (cultivars), precise fertilization, increase of soil moisture, and chemical treatments. Yet, these management practices do not have common or reproducible results at differing sites. A monitoring study was done in 32 sites to evaluate the relation between CS and biological/chemical soil parameters. Correlations were observed between scab severity and content of nutrients such as Fe, N, and Ca in soil and periderm, and between disease severity and abundance of actinobacteria and total bacteria, together with the pathogenicity determinant, txtB gene (biosynthetic gene of thaxtomin) in both soil and periderm of potatoes. The findings led to novel conclusions, which can help to understand relationships applicable in scab control. Peat and DTPA chelated iron were supplemented to pots filled with soil conducive for CS in order to determine the effects of soil organic matter, iron and pH on CS development. The results were compared with data obtained for a suppressive soil from a nearby field with naturally low CS severity. Both peat and iron supplements decreased CS and the combination...
The genetic basis of Czech garlic "paličák" (A. sativum L. ssp. ophioscorodon) focusing on the biosynthetic pathways of the secondary metabolites
Čermák, Vladimír ; Ovesná, Jaroslava (advisor) ; Demnerová, Kateřina (referee)
Czech bolting garlic (Allium sativum L.) is the name for Czech varieties, which are categorized into subspecies sativum ssp. An inflorescence production and specific onion morphology is typical for this type of garlic. The genetic basis, that has been described by the analysis of microsatellites, are discussed in this thesis. Transcription analysis has revealed a large number of unigenes that have been assigned by genetic ontology to individual functions in the organism. Therefore, genetic differences from other varieties were confirmed. The impact of the environment and cultivation practices, including large- scale production, is being explored. The results can be used for further research or breeding. Consumers and experts have rated the Czech bolting garlic as a variety with a distinctly pungent taste and aroma. Substances causing this strong characteristic taste are secondary metabolites, alk(en)ylcysteine-S-oxides (ACSO), especially alliin and methiin. Chemical analysis did not confirm the exceptionally high value of these taste precursors or other substances that should distinguish Czech bolting garlic. Secondary metabolism includes other important substances that are used mainly in pharmacy and healthcare, where garlic has been used for thousands of years due to antibacterial effects,...
Study of biofilm formation of bacterial isolates from milk and dairy products
Nováková, Veronika ; Demnerová, Kateřina (advisor) ; Seydlová, Gabriela (referee)
Bacterial biofilm is a complex community of microbial cells, which are embed into the extracellular pollysacharide matrix. Typical ability of the biofilm is the ability to adhere to either abiotic, or biotic surfaces. The formation of biofilm is a dynamic process, which finally formes a 3D multicellular complex. The initial phase is called the adhesion. After that process is done, the extracellular polymeric matrix is produced to create living conditions of bacteria in biofilm. Biofilm has hight level of antimicrobial resistance. This resistance consists of physical and chemical barriers, which effectively block diffusion and penetration of antimicrobial substances inside the biofilm. Biofilms cause problems in food industry, where it could be origin of food contaminations. Thefore it is necessary to understand the relationship between microorganisms and materials used in food industry. Key words: biofilm, milk and dairy products, milk and dairy industry, disifectanc, Escherichia coli, Staphylococcus spp.
Differentiation in Saccharomyces cerevisiae colonies
Čáp, Michal ; Palková, Zdena (advisor) ; Pichová, Iva (referee) ; Demnerová, Kateřina (referee)
Yeast colonies are, alongside to bacterial biofilms, multicellular communities formed by unicellular microorganisms. These specific communities differ in many ways from populations cultivated in planktonic cultivations. Gradients of nutrients, metabolic by- products and other factors are formed and preserved within these structures, which provides a basis for cellular differentiation. Current literature concerning these issues with emphasis on yeast colonies and biofilms is summarised in the Introduction of this work. Section Results then describes my contribution to the knowledge on the differentiation of the colonies of Saccharomyces cerevisiae as a model system for studying microbial multicellular structures. Previously described horizontal differentiation, i.e. differentiation between colony centre and margin, is further characterised with respect to ammonia signalling and stress resistance. The importance of genes conferring the cell`s oxidative stress resistance in colony differentiation was studied and it was concluded that not stress resistance but rather metabolic and other changes promoted by ammonia signal are important for colony differentiation and survival. A new type of colony differentiation - the horizontal, i.e. differentiation between upper and lower part of the colony, is...
Differentiation of yeast colonies and development of new approaches to monitor oxygen and nutrient availability
Vopálenská, Irena ; Janderová, Blanka (advisor) ; Demnerová, Kateřina (referee) ; Pichová, Iva (referee)
Yeast Saccharomyces cerevisiae as an unicellular organism is one of the best-studied experimental organisms. It is an important model organism for the study of intracellular processes of eukaryotic cells. Yeasts are also social organisms with cell-to-cell communication able to form organized multicellular structures (colonies and biofilms). Yeast and other microorganisms in nature prefer to form colonies on solid substrates rather than to grow as "planktonic" single cells (Palková, 2004; Wimpenny, 2009). The yeast S. cerevisiae typically forms colonies, biofilms were described only rarely. Yeast colonies exhibit an organized morphological pattern characteristic of each particular yeast strain (Kocková-Kratochvílová, 1982). This work is focusing on morphology and differentiation of the S. cerevisiae colonies of common laboratory strains forming less structured colonies, and strains of the Σ1278b genetic background forming highly structured "fluffy" colonies. It shows that polarized budding pattern and especially cell ability to form aggregates enable development of structured morphology. During development of "fluffy" colonies two differently regulated events of dimorphic switch from yeast form to filamentous growth occur. One of these events is dependent on the surface glycoprotein, Flo11p flocculin. This...
The Introduction of Contact Ecotoxicity Tests for the Assessment of Terrestrial Ecosystems
Modlitbová, Pavlína ; Kráčmar, Stanislav (referee) ; Beklová, Miroslava (referee) ; Demnerová, Kateřina (referee) ; Vávrová, Milada (advisor)
This dissertation thesis is focus on using toxicity tests with testing organism - terrestrial isopod Porcellio scaber. This organism is use for toxicity assessment of selected anorganic compunds which could contaminate terrestrial ecosystem. This organism was selected for several reasons - sophisticated test methodology; well known biology of these animals and their ecological relevance. Besides classical endpoints such as mortality, change in mass of test specimens and the effect of the presence and concentration of contaminants onto food consumption; we monitored changes in the morphometric characteristics at the level of tissues and cells, the cytotoxicity and possibly bioaccumulation in various parts of the body of the organism. Selected test substance in this thesis were nanoparticles and inorganic salts. Wherein for chosen gold nanoparticles was carried out a complete study presents several levels of organization of living matter (organism, tissue, cell). For selected salts compound, this study was carried out on the behavior of organisms and classical endpoints.
Application of Separation Techniques Connected with Mass Spectrometry for the Analysis of Environmentally Important Compounds
Mácová, Daniela ; Čelechovská, Olga (referee) ; Márová, Ivana (referee) ; Demnerová, Kateřina (referee) ; Čáslavský, Josef (advisor)
The identification of the hydrolysis and photodegradation products of flexible polyurethane foams (PUFs) with addition of biooriginated and biodegradable additive was the first topic of this dissertation work. Separation of polyurethane foam hydrolysis degradation products, designed for ecotoxicological tests, was managed by high-performance liquid chromatography coupled with mass spectrometry (HPLC/MS). The degradations product structure was elucidated by tandem mass spectrometry (MSn). PUF photodegradation products were obtained by exposure of materials by irradiation at 254 nm. Semi-volatile degradation products were isolated from the exposed polyurethane by n-hexane extraction; volatile compounds were collected by solid phase microextraction (SPME). Gas chromatography with mass spectrometry (GC/MS) and complete orthogonal tandem gas chromatography with mass spectrometry (GCxGC/TOF MS) was used for separation and identification of photodegradation products. The influence of the bio-filler on the character of degradation products and the possible effect of PUF degradation products on the environment was discussed at the end of this section. The determination of isoprostanes – markers of oxidative stress in tissues of beadlet anemone (Actinia equina) was the subject of the second topic. F2-isoprostanes were synthesized from the arachidonic acid. With thereby prepared isoprostanes the method of determination by liquid chromatography with tandem mass spectrometry (HPLC/MS/MS) was developed and optimized. The isoprostane isolation process from the Actinia equina tissues was optimized with solid phase extraction (SPE). The resulting methodology was used to quantify isoprostanes in tissues of anemones, which were exposed to both moderate and high temperature changes. The temperature changes were used to initiate the oxidative stress in organisms. In addition, concentration levels of unknown compounds were also monitored. These unknown compounds were extracted from tissues together with F2-isoprostanes and their identity is discussed in this dissertation work too. The possibility of using isoprostane levels in the Anthozoa tissues for the oxidative stress monitoring is discussed in the conclusion of this work.
Use of Waste Substrates to Production of Enriched Yeast Biomass
Starečková, Terezie ; Demnerová, Kateřina (referee) ; Vávrová, Milada (referee) ; doc.PharmDr.Petr Babula, Ph.D. (referee) ; Márová, Ivana (advisor)
Yeasts are like other organisms constantly exposed to environmental influences. Their survival depends on the skills to adapt to environmental changes, including the ability to use various alternative sources of nutrients. In presented PhD thesis carotenogenic yeast belonging to the genera Rhodotorula, Sporobolomyces and Cystofilobasidium were tested for ability to use of selected waste substrates, and also subjected to several types of exogenous stress effects and mutations in order to increase the production of microbial biomass enriched with specific metabolites. As alternative nutrient sources derived from waste substrates from agricultural and farm production apple peel, pulp, corn germ and more were tested. Yeasts were also exposed to osmotic, oxidative and combined stress (benefits of various concentrations of NaCl and H2O2 to the culture media), followed by metal ions of selenium and chromium in concentrations of 0.01 mM, 0.1 mM and 1 mM. The effect of mutagen methanesulfonic acid ethyl ester was tested too. In all experiments the adaptivity of cells, morphological changes, color pigments produced by the media while some important fungal metabolites production and changes in chromosomal DNA fragmentation were analyzed. In order to evaluate potential changes in the yeast genome after treatment with mutagen and stress factors methods for isolation of intact chromosomal DNA and DNA analysis by pulsed field gel electrophoresis was optimized. The amount of produced metabolites was mainly analyzed by RP-HPLC with UV/VIS and MS detection. The work has been shown that most strains are able to use waste substrates and produced selected target metabolites. Biomass, for example, in R. aurantiaca on apple fiber was about 7 g/l and in C. capitatum cultivated on modified whey reached to 9 g/l. Amount of produced carotenoids by R. aurantiaca cultivated on wheat germ and maize after enzymatic hydrolysis by F. solani was 1.01 mg/g and S. roseus on pasta 4.3 mg/g. The values of ergosterol synthesis in R. aurantiaca are on the apple shells around 4.8 mg/g, in S. roseus on pasta with the enzymatic hydrolysis of P. chrysosporium 8.9 mg/g. The best substrate for biomass production and induction of carotenoids are waste substartes containing a mixture of simple and complex carbohydrates enriched with the addition of nitrogen compounds. Potential cytotoxic effect of stress factors of low concentrations was demonstrated. Red yeast genome was able to distribute by optimized PFGE, the karyotype of tested yeasts contain 11 or more chromosomes with visible differences between yeast species and genera. During exchange internship the ability of recombinant yeast S. cerevisiae to convert xylose to xylitol, which would be achieved by increasing the production of bioethanol as alternative fuel sources was studied. It turned out that both ligninocellulose materials to bioethanol production, as well as various waste substrates for microbial synthesis of carotenoids would reduce costs for industrial production of yeast metabolites, as well as to reduce the negative burden on the environment.
Controlled Production and Degradation of Selected Biomaterials
Obruča, Stanislav ; Demnerová, Kateřina (referee) ; Vávrová, Milada (referee) ; Němec, Miroslav (referee) ; Márová, Ivana (advisor)
Předložená disertační práce se zabývá studiem produkce a degradace polymerních materiálů s využitím mikroorganismů. Hlavní pozornost je upřena ke studiu produkce polyesterů bakteriálního původu - polyhydroxyalkanoátů. Tyto materiály jsou akumulovány celou řadou bakterií jako zásobní zdroj uhlíku, energie a redukční síly. Díky svým mechanickým vlastnostem, kterými silně připomínají tradiční syntetické polymery jako jsou polyetylén nebo polypropylén, a také díky své snadné odbouratelnosti v přírodním prostředí, jsou polyhydroxyalkanoáty považovány za ekologickou alternativu k tradičním plastům vyráběným z ropy. Polyhydroxyalkanoáty mají potenciál najít řadu aplikací v průmyslu, zemědělství ale také v medicíně. Významná část předložené práce je zaměřena na produkci polyhydroxyalkanoátů z odpadních substrátů pocházejících především z potravinářských výrob. Testována byla odpadní syrovátka nebo odpadní oleje z různých zdrojů. Právě využití levných odpadních substrátů je strategií, která by mohla přispět ke snížení ceny polyhydroxyalkanoátů a tím usnadnit jejich masové rozšíření. Podle výsledků dosažených v této práci jsou právě odpadní olejové substráty velice perspektivní cestou k ekonomicky rentabilní biotechnologické produkci polyhydroxyalkanoátů. Další část předložené práce se zabývá studiu spojení metabolické role polyhydroxyalkanoátů a stresové odpovědi bakterií. V této práci bylo zjištěno, že expozice bakteriální kultury řízené dávce etanolu nebo peroxidu vodíku významně navýší dosažené výtěžky a to o přibližně 30 %. Po aplikaci výše zmíněných stresových faktorů došlo k aktivaci metabolických drah vedoucí k odbourání stresového faktoru z média. Výsledkem bylo navýšení poměru NAD(P)H/NAD(P)+, což vedlo k částečné inhibici Krebsova cyklu a naopak aktivaci biosyntetické dráhy polyhydroxyalkanoátů. Mimoto došlo k významnému navýšení molekulové hmotnosti výsledných materiálů. Podle těchto výsledků se regulovaná aplikace vhodně zvolených stresových podmínek zdá být zajímavou strategií, která vede nejen k navýšení celkových výtěžků, ale také významnému zlepšení vlastností polymeru. Poslední část disertační práce se zabývala studiem procesu biodegradace polyuretanových materiálů. Polyuretanové eleastomery byly modifikovány rozličnými biopolymery za účelem navýšení jejich biodegradability. Tyto materiály byly posléze vystaveny působení směsné termofilní kultury jako modelového systému, který simuluje přirozené konsorcium bakterií. Přítomnost testovaných materiálů v kultivačním médiu vedla k neobvyklým růstovým charakteristikám bakteriální kultury. V průběhu prvních několika dní byl růst kultury silně inhibován, nicméně po překonání této neobvykle dlouhé lag-fáze došlo k intenzivnímu nárůstu kultury. Hlavní podíl na hmotnostním úbytku testovaných materiálů během experimentů měl samovolný rozpad materiálů, nicméně byl pozorován i vliv bakteriální kultury, kdy míra biotické degradace závisela na použitém modifikačním činidle. Nejvyšší míra biotické degradace byla pozorována u polyuretanového materiálu modifikovaného acetylovanou celulózou. Lag-fáze byla způsobena uvolněním nezreagovaného katalyzátoru (dibutylcínlaurát) a polyolu do kultivačního média. Bakteriální kultura se však po čase dokázala na přítomnost toxických látek v médiu adaptovat nebo je dokázala eliminovat.

National Repository of Grey Literature : 35 records found   1 - 10nextend  jump to record:
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