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Interaction of non-thermal plasma at atmospheric pressure (CAPP) with non-pathogenic bacteria
Chobotská, Barbora ; Brázda, Václav (referee) ; Kozáková, Zdenka (advisor)
The subject of this bachelor thesis is the study of decontamination effects of cold atmospheric pressure plasma (CAP) on selected bacteria Escherichia coli and Staphylococcus epidermidis. A non-thermal microwave plasma torch was used. The plasma torch was connected to the software in the computer, which allowed its movement over the treated area. The used power fluctuated between 12–13 W and argon (gas purity 4.6) was chosen as the working gas with a constant flow rate of 5 l/min Inhibition efficacy was observed depending on the type of the selected treatment. The chosen treatment parameters included the effect of speed, direction, and treatment time for both studied bacteria. The aim was to achieve the highest decontamination of the treated area and to determine which of these parameters appeared to be the most significant. No obvious difference (between the treatment directions) was found in the overall decontamination of the treated area for studied bacteria Escherichia coli. The negative effect of speed was only observed for the fastest speed used in the case of gram-positive Staphylococcus epidermidis. The most significant parameter was found to be the treatment time, where a significant decrease in colony growth was observed with increasing treatment time. Furthermore, a repeated treatment was done, where the inoculum was prepared from the already treated bacteria. By this set of experiments, possible development of microbial resistance against the plasma treatment was tested. The results showed that there was no significant increase in the number of colonies even after the repeated treatment. It was also observed that the gram-positive bacteria Staphylococcus epidermidis showed lower decontamination effect evaluated via the number of colonies than the gram-negative bacteria Escherichia coli for all treatment types studied.
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Impact of honey enrichment on its antimicrobial activity by bioactive substances against Escherichia coli
ŠEBESTOVÁ, Lucie
The main aim of this bachelor's thesis is to determine the Minimum Inhibitory Concentration (MIC) and the Minimum Bactericidal Concentration (MBC) of both native honeys and honeys enriched with selected essential oils on the growth of Escherichia coli. In the theoretical part of the bachelor's thesis, bacteria Escherichia coli is briefly characterized, including its pathogenic strains and antibiotic resistance. Furthermore, honey is described, including its chemical composition and physiological effects. Additionally, essential oils are generally described, including their antimicrobial properties, along with a brief description of each selected oil. In the methodological part of the bachelor's thesis, procedures for preparing mediums, agar, inoculum, and the determination of their minimum inhibitory and bactericidal concentrations are described. MIC was measured spectrophotometrically in 96 well microtiter plates. MBC was determined by inoculation from the plate onto Mueller-Hinton agar followed by a 24hour incubation. A total of 3 native honeys were tested, which were then enriched with 8 selected essential oils. Essential oils with the addition of dimethyl sulfoxide (DMSO) were also tested. The results of the obtained data are statistically evaluated, graphically presented, and compared with scientific sources.
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Preparation of skin masks with the anti-acne effect
Geierová, Kateřina ; Bendová, Agáta (referee) ; Márová, Ivana (advisor)
This bachelor thesis was focused on the preparation and characterization of nanofibers containing active agents with the aim of using these materials in anti-acne facial masks. The nanofibers were prepared from polyhydroxybutyrate solution and gelatin solution by electrospinning method. Among the active ingredients, eugenol and piperine were used to functionalize the nanofibers. In the theoretical part, the types of face masks, the individual active ingredients, the materials for the preparation of nanofibers, their preparation methods and their characterization were characterized. In the practical part, the antioxidant and antimicrobial activity of the active ingredients themselves was investigated. Subsequently, the release rate of selected active ingredients from the prepared nanofibers was monitored in three model environments. The antimicrobial activity of the nanofibers was monitored on three microorganisms, namely Staphylococcus epidermidis, Cutibacterium acnes and Escherichia coli.
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Biological effects of substances isolated from Isoptera species
Dušková, Simona ; Hrstka, Miroslav (referee) ; Brázda, Václav (advisor)
This thesis was focused on monitoring the viability of eukaryotic and prokaryotic cells after exposure of termites-isolated chemicals. Recently, evidence of antibacterial and antifungal properties of these defense substances has grown, and they can find a wide range of uses not only in the pharmaceutical industry. In this work, three defensive substances from termites were studied: nerolidol, nitropentadecene and methylanthranilate. Their antibacterial effects, minimal inhibitory concentrations and minimal bactericidal concentrations against Escherichia coli STBL3 strain were monitored. Further, their cytotoxic effects on eukaryotic non-tumor (HEK293FT) and tumor cells (MCF7) as well as their effect on plasmid DNA were studied. Antibiotic ampicillin and cytostatic cisplatin were used as control substances for antibacterial and cytotoxic effects, respectively. In the case of the action of nerolidol, nitropentadecene and methylanthranilate on the STBL3 strain, antibacterial activity was not demonstrated. Cytotoxic effects were observed nerolidol and nitropentadecene. None of the examined substances modified the plasmid DNA.
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Study of elementary inactivation processes acting during sterilization of procaryotic systems in dielectric barrier discharge
Bittnerová, Zuzana ; Sťahel, Pavel (referee) ; Kozáková, Zdenka (advisor)
Presented master’s thesis is focused on the study of the elementary inactivation processes acting during sterilization of procaryotic systems in dielectric barrier discharge (DBD). Sterilization is an important biomedical and food-industry application and plasma sterilization is one of the methods, which are suitable for sterilization of heat and chemical sensitive materials. Biologically contaminated samples were treated in dielectric barrier discharge operated at atmospheric pressure. The discharge was generated in argon and in nitrogen. The plasma power density was 2725,93 mW cm–3 in argon and 2325,93 mW cm–3 in nitrogen. Gram positive bacteria Bacillus subtilis and gram negative bacteria Escherichia coli were used as a bioindicator. Bacteria were spread onto the surface of Whatman No.1 filtration paper. The influence of UV radiation, reactive species, heat and plasma discharge where the synergistic function of all of the agents was studied. Effects of UV radiation and temperature were studied separately. In order to separate the effect of UV radiation generated by DBD the quartz glass transmitting UV radiation was employed. During the plasma exposition selected samples were covered with the quartz window while other samples were directly exposed to the plasma. Covered samples were exposed to UV radiation and temperature (which cannot be eliminated), samples without quartz window were directly plasma exposed (treated). Results show that for covered samples the lower inactivation was reached than by the samples directly exposed to plasma. When studying the effect of temperature, the temperature between the DBD electrodes was measured by means of a thermocouple. Afterwards the samples were placed in an oven and exposed to the same temperature as was measured between the electrodes. By comparing the results of heat treated samples and plasma treated samples it can be assumed that the influence of the temperature during the sterilization process in DBD is very low. The discharge parameters were studied by means of the Optical Emission Spectroscopy. Plasma treated samples were assessed employing Scanning Electron Microscopy (SEM). Damage of Bacillus subtilis cell wall due to the effect of plasma was observed while no effect of plasma on the structure of filtration paper was detected.
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Preparation of edible antimicrobial packaging
Kozubová, Petra ; Dzurická, Lucia (referee) ; Skoumalová, Petra (advisor)
This bachelor’s thesis is focused on preparation and characterization of eatable packaging. Eatable packaging consists of two components, antimicrobial component and base of packaging. The task of eatable packaging is to protect product against antimicrobial attack and to extend durability of the product. As antimicrobial components were used water and oil extracts of several herbs (mint, nettle, lavender, cinnamon, cloves, vanilla, ginger and rhinoceros). Oil extracts were also encapsulated into liposomes for better application. Firstly, concentrations of polyphenoles and antioxidants of extracts were determined. Oil extract of clove contained the highest concentration of both determined substances. Encapsulation efficiency, stability and size of liposomes were tested too. All prepared liposomes were stable and relatively of the same size. Also high encapsulation efficiency was observed. Next, antimicrobial activity of prepared extracts and liposome particles against yeast Candida glabrata and two bacteria strains Escherichia coli and Staphycococcus epidermidis was tested and the high antimicrobial activity reported mainly all tested liposomes. Then alginate, chitosan and carboxymethylcellulose and their combinations respectively were used as bases of packaging. Created film had to meet several parameters, especially suitable sensory properties such as pleasant taste and smell, as well as invisibility, gloss, affordability and suitable manipulability. Combination of 2% carboxymethylcellulose and 1% alginate in a ration of 2:1 with liposomes of mint as antimicrobial component achieved the best results.
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Electrophotocatalytic wather disinfection
Štefancová, Eva ; Drbohlavová, Jana (referee) ; Veselá, Mária (advisor)
This work focuses on the photoelectrocatalytic water disinfection by applying electrical voltage onto an electrode system. Working electrode consist of electrical conductor covered by thin film TiO2, because of its very important photoactivity. Disinfection effect was monitored on Gram-negative bacteria E. coli under different conditions. Inactivation of bacterial cells in contamined aqueous solution, first experiment was observed only with presence of oxygen, later reactor was irradiated with UV-A compact blacklight with intensity of 4 mW• m-2 or electrical voltage of 1V was applied to electrode. By adding both effects at the same time I was verifying photoelectrocatalysic water disinfection on chosen microorganism.
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Study of plasma interaction with bacteria for wound healing
Šrámková, Sarah ; Krčma, František (referee) ; Kozáková, Zdenka (advisor)
This thesis focuses on the study of the interaction of plasma with bacteria to enhance the promotion of wound healing. Firstly, the wound healing process is described, followed by low temperature plasma, its effect on bacterial cells and its use in biomedicine. The experimental part deals with the application of low-temperature plasma generated by microwave jet on selected microorganisms and the influence of experimental conditions on the antimicrobial effect of the plasma. One representative of the microorganisms occurring in the wounds was selected as representatives of Gram-positive bacteria and Gram-negative bacteria, namely Staphylococcus epidermidis and Escherichia coli. Using the results obtained, the antimicrobial effect of plasma generated by microwave jet in argon was confirmed. The degree of antimicrobial effect is related to the concentration of the microorganisms, the time of treatment and whether the bacteria are Gram-positive or Gram-negative.
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Preparation of some application formula of antimicrobial substances
Krempaský, Kamil ; Němcová, Andrea (referee) ; Márová, Ivana (advisor)
The main aim of this study is to determine antimicrobial activity of herbal extracts and also encapsulate these extracts and lysozyme to liposomes. The extracts were prepared from above ground part of wormwood (Artemisia absinthium), aerial part of breckland thyme (Thymus serpyllum), the root of purple coneflower (Echinacea purpurea), liquorice (Glycyrrhiza glabra), corolla of pot marigold (Calendula officinalis) and berries of sea buckthorn (Hippophae rhamnoides). In all extracts antioxidant activity and the amount of total polyphenols and flavonoids was determined. All of these extracts and also lysozyme were used for encapsulation to liposomes. Prepared liposomes were then tested for their stability. Further, encapsulated as well as free extracts were tested for their antimicrobial activity on Escherichia coli, Serratia marcescens, Micrococcus luteus and Bacillus subtilis. Extract from Echinacea purpurea showed the highest antimicrobial activity.
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Study of Sterilization Effects Initiated by Dielectric Barriere Discharge
Slámová, Jitka ; Pekárek, Stanislav (referee) ; doc. RNDr. Karol Hensel, Ph.D., oponent (referee) ; Krčma, František (advisor)
The overall goal of the presented dissertation thesis was to study the sterilization efficiency of dielectric barrier discharge operated at atmospheric pressure. The fungi Aspergillus niger, gram-positive bacteria Bacillus subtilis and in some experiments also gram-negative bacteria Escherichia coli were used as a bio-indicator enabling to evaluate the effect of plasma assisted microbial inactivation. The samples of microorganism were placed on paper Whatman 1 or PET foil and exposed to plasma. The plasma was generated in argon, nitrogen, synthetic dry/humid air with frequency up to 10 kHz and plasma power density in the range of 1,2-2,9 W/cm3 (according to the process gas). The influence of process gas, plasma power density, plasma exposition time, type of microorganism and material of the substrate on the sterilization effect of dielectric barrier discharge was evaluated. Furthermore the contribution of each single mechanism (UV radiation, temperature and reactive species) to the sterilization effect of plasma and influence of gas humidity was evaluated. The DBD was analysed by means of optical emission spectroscopy, thermocouple was used to measure temperature during a sterilization process. In order to verify the mechanical damage of the microbial cell or the substrates during the plasma process the samples were studied by scanning electron microscopy. Generally, on the basis of experimental results, at increasing treatment times, the remaining number of spores (CFU) decreased. Similarly at increasing the plasma power input, the sterilization rate increased. When sterilising the spores of A. niger in plasma using different process gasses, the efficiency of plasma sterilization decreased as follows: argon, humid synthetic air, nitrogen and dry synthetic air. The results observed in argon plasma using different microorganism demonstrated that the sensitivity of vegetative cells resp. spores to DBD decreased as follows: A. niger spores, B. subtilis vegetative cells, E. coli vegetative cells and B. subtilis spores. Simultaneously results observed for sterilization of spores and vegetative cells of B. subtilis and A. niger demonstrated that the spores are generally more resistant to plasma than are the corresponding vegetative cells. Combining the results of contribution of each single mechanism, optical emission spectroscopy and inactivation characteristic it was found out that the reactive species significantly contribute to the plasma sterilization in all process gasses. Furthermore the inactivation process can be partly assisted by UV radiation and also the temperature can contribute in limited extent to inactivation process in some gasses. The contribution of UV radiation to the plasma sterilization decreased as follows: nitrogen, argon, dry syntetic air and humid syntetic air. Moreover it was found out that the contribution of each single mechanism can be species dependent, this is due to the different response of microorganism to the unfavorable external conditions. SEM analysis of the substrates prooved the etching actions of the plasma generated in all process gasses on the surface of the PET foil. The several minute plasma exposition of the PET foil resulted in the occurence of the „hole corrosion“ on the PET surface. Contrary to these there were no visible changes observed in the paper structure.
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