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Studium účinků plazmatem ošetřených kapalin na zvolené mikroorganismy
Klementová, Kamila ; Jirásek, Vít (oponent) ; Trebulová, Kristína (vedoucí práce)
Předmětem bakalářské práce je stanovení účinků plazmatem ošetřených kapalin na bakterie Staphylococcus aureus a Pseudomonas aeruginosa. Rezistence bakterií vůči antibiotikům je velmi diskutovaným tématem dnešní doby. Plazmatem ošetřené kapaliny mají potenciál k řešení tohoto problému díky svým antimikrobiálním účinkům. Tyto účinky však dosud nebyly úplně objasněné, a proto jsou předmětem této práce. Teoretická sekce je zaměřena hlavně na mikrobiologickou část, konkrétně na morfologii bakteriální buňky a jednotlivé bakterie použité v experimentální části. Dále je zde popsáno plazma, nízkoteplotní atmosférické plazma (CAP) a interakce plazmatu s kapalinou. Poslední část teoretické části se zabývá potenciálním využitím nízkoteplotního plazmatu a plazmatem ošetřených kapalin v oblasti potravinářského a zemědělského průmyslu a zdravotnictví. Plazmatem ošetřené kapaliny (PTL) byly v experimentální části připraveny z fyziologického roztoku dvěma plazmovými systémy. Byly také připraveny roztoky simulující složení připravených PTL a standardní roztoky dusitanů, dusičnanů a peroxidu vodíku. Účinky plazmatem ošetřených kapalin byly porovnány s účinky roztoků simulujících složení plazmatem ošetřených kapalin a se standardními roztoky. Testování účinků probíhalo na standardizovaných bakteriálních kulturách Staphylococcus aureus a Pseudomonas aeruginosa. Jejich suspenze byly vystaveny ošetřeným kapalinám, připraveným standardním roztokům a alternativám PTL. Ve zvolených expozičních časech bylo provedeno stanovení počtu životaschopných bakterií očkováním na živnou půdu a po 24-hodinové inkubaci byl vyhodnocen antimikrobiální účinek počítáním kolonie tvořících jednotek s využitím softwaru Aurora. Výsledky této práce dokázaly lepší antimikrobiální účinnost plazmatem ošetřených kapalin než roztoků simulujících jejich složení. Byl zjištěn rozdíl v účincích mezi dvěma různě připravenými plazmatem ošetřenými kapalinami. Bylo také dokázáno, že bakterie Pseudomonas aeruginosa je citlivější na expozici plazmatem ošetřených kapalin než bakterie Staphylococcus aureus.
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Study of plasma interaction with yeast Candida glabrata
Trebulová, Kristína ; Lehocký, Marián (oponent) ; Krčma, František (vedoucí práce)
The main subject of this diploma thesis is the study of low temperature discharges´ effects on the yeast Candida glabrata (C. glabrata). Cold atmospheric-pressure plasma (CAPP) technology has achieved a great deal of attention in recent years, due to its positive attributes, including high efficiency against a wide range of microorganisms, ease of operation, economic simplicity and also environmental friendliness. The research of CAPP focuses mainly on the applications involving the treatment of cancer and superficial infections as well as sterilization technologies in medicine, food and agriculture. Various effects and also some mechanisms of inactivation of bacteria and tumor cells have been explained over the last decade but the effects and mechanisms of fungi inactivation have not yet been sufficiently investigated. The aim of this work is to verify the effects of chosen low-temperature discharges on the model yeast example C. glabrata, in order to establish basic plasma set-up for further studies and experiments in vivo and to expand a global knowledge about the antifungal effects of CAPP. In this study, 5 different discharges, namely: microwave (unipolar and surface wave) in the form of plasma torch, transient spark, corona discharge in the form of corona pen and dielectric barriere discharge (DBD) in the form of DBD pen were tested. All discharges were tested by direct application onto a 24-hour inoculum of C. glabrata inoculated on agar plates. The results were evaluated based on the size and stability of the inhibition zones formed in the post treatment cultivation. From the individual measurements optimal conditions for testing the inhibitory effects of low temperature discharges on the selected yeast were determined. The microwave discharges (unipolar and surface wave) were chosen to be the most effective in the inhibition of C. glabrata. The measurements were therefore pursued with these discharges under selected conditions corresponding to chosen application of sterilization of surface infections in living organisms. The unipolar microwave discharge was chosen to be the most effective discharge for the potential treatment of superficial candidiasis. The indirect effect of plasma was also investigated using plasma activated liquid (PAL) on inoculated yeast culture. PAL was prepared using 3 different discharges: Dielectric Barrier Discharge (DBD), Unipolar Microwave Discharge and Electric Discharge in Liquid. The results of this work prove significant inhibitory effects of CAPP on the yeast C. glabrata, within the limits of in vitro cultivation and the so-called flat biology. The overall inhibitory effects are directly proportional to the irradiation time, the magnitude of applied power and also the area covered by the discharge. In terms of the initial cell concentration an indirect dependency can be observed, but it can also be stated that this factor is less significant compared to the other examined factors. Observations of the effects of plasma activated liquid (PAL) indicate that a single application of PAL under selected conditions is not alone enough for successful inhibition of C. glabrata.
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Study of plasma jet effects on yeast
Trebulová, Kristína ; Skoumalová, Petra (oponent) ; Krčma, František (vedoucí práce)
The purpose of this work is to study the effects of a 2.45 GHz microwave plasma jet on the yeast Candida glabrata. The microwave plasma was generated by a surfatron resonator and as a working gas pure argon at constant flow rate 5 Slm was used. Thanks to a high number of active particles generated in a low temperature plasma, this type of plasma has become highly popular, especially thanks to its bactericidal effects. However, its antifungal effects and mechanisms of fungal inactivation have still not been fully understood. Therefore, the study focuses on the antimycotic effects of the low temperature plasma in a form of a microwave discharge on Candida glabrata as a model yeast example. The principal focus is on the measurement and evaluation of changes in inactivation effects caused by varying the initial concentration of Candida glabrata cells, applied power, and the exposure time. The theoretical part presents the basic information concerning Candida glabrata, its pathogenicity, virulence and infectivity. Furthermore, plasma in general, various types of discharges and their applications in the food and biomedical industries are discussed. The experimental part focalizes on the determination of the inactivating effects of the microwave plasma jet on the yeast Candida glabrata. The experiment consists of applying the discharge on a freshly inoculated colony of Candida glabrata on agar plates and observing inhibitory effects in the form of inhibitory zones formed after subsequent cultivation. The inhibition of yeasts by the microwave discharge is interesting scientifically due to its promising results, which have also been proven in this work, but also economically, especially due to the use of argon. Argon as a carrier gas is much cheaper than helium and colder than common atmospheric pressure discharges in air such as DBDs, corona or glow discharges.
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Study of plasma interaction with yeast Candida glabrata
Trebulová, Kristína ; Lehocký, Marián (oponent) ; Krčma, František (vedoucí práce)
The main subject of this diploma thesis is the study of low temperature discharges´ effects on the yeast Candida glabrata (C. glabrata). Cold atmospheric-pressure plasma (CAPP) technology has achieved a great deal of attention in recent years, due to its positive attributes, including high efficiency against a wide range of microorganisms, ease of operation, economic simplicity and also environmental friendliness. The research of CAPP focuses mainly on the applications involving the treatment of cancer and superficial infections as well as sterilization technologies in medicine, food and agriculture. Various effects and also some mechanisms of inactivation of bacteria and tumor cells have been explained over the last decade but the effects and mechanisms of fungi inactivation have not yet been sufficiently investigated. The aim of this work is to verify the effects of chosen low-temperature discharges on the model yeast example C. glabrata, in order to establish basic plasma set-up for further studies and experiments in vivo and to expand a global knowledge about the antifungal effects of CAPP. In this study, 5 different discharges, namely: microwave (unipolar and surface wave) in the form of plasma torch, transient spark, corona discharge in the form of corona pen and dielectric barriere discharge (DBD) in the form of DBD pen were tested. All discharges were tested by direct application onto a 24-hour inoculum of C. glabrata inoculated on agar plates. The results were evaluated based on the size and stability of the inhibition zones formed in the post treatment cultivation. From the individual measurements optimal conditions for testing the inhibitory effects of low temperature discharges on the selected yeast were determined. The microwave discharges (unipolar and surface wave) were chosen to be the most effective in the inhibition of C. glabrata. The measurements were therefore pursued with these discharges under selected conditions corresponding to chosen application of sterilization of surface infections in living organisms. The unipolar microwave discharge was chosen to be the most effective discharge for the potential treatment of superficial candidiasis. The indirect effect of plasma was also investigated using plasma activated liquid (PAL) on inoculated yeast culture. PAL was prepared using 3 different discharges: Dielectric Barrier Discharge (DBD), Unipolar Microwave Discharge and Electric Discharge in Liquid. The results of this work prove significant inhibitory effects of CAPP on the yeast C. glabrata, within the limits of in vitro cultivation and the so-called flat biology. The overall inhibitory effects are directly proportional to the irradiation time, the magnitude of applied power and also the area covered by the discharge. In terms of the initial cell concentration an indirect dependency can be observed, but it can also be stated that this factor is less significant compared to the other examined factors. Observations of the effects of plasma activated liquid (PAL) indicate that a single application of PAL under selected conditions is not alone enough for successful inhibition of C. glabrata.
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Study of plasma jet effects on yeast
Trebulová, Kristína ; Skoumalová, Petra (oponent) ; Krčma, František (vedoucí práce)
The purpose of this work is to study the effects of a 2.45 GHz microwave plasma jet on the yeast Candida glabrata. The microwave plasma was generated by a surfatron resonator and as a working gas pure argon at constant flow rate 5 Slm was used. Thanks to a high number of active particles generated in a low temperature plasma, this type of plasma has become highly popular, especially thanks to its bactericidal effects. However, its antifungal effects and mechanisms of fungal inactivation have still not been fully understood. Therefore, the study focuses on the antimycotic effects of the low temperature plasma in a form of a microwave discharge on Candida glabrata as a model yeast example. The principal focus is on the measurement and evaluation of changes in inactivation effects caused by varying the initial concentration of Candida glabrata cells, applied power, and the exposure time. The theoretical part presents the basic information concerning Candida glabrata, its pathogenicity, virulence and infectivity. Furthermore, plasma in general, various types of discharges and their applications in the food and biomedical industries are discussed. The experimental part focalizes on the determination of the inactivating effects of the microwave plasma jet on the yeast Candida glabrata. The experiment consists of applying the discharge on a freshly inoculated colony of Candida glabrata on agar plates and observing inhibitory effects in the form of inhibitory zones formed after subsequent cultivation. The inhibition of yeasts by the microwave discharge is interesting scientifically due to its promising results, which have also been proven in this work, but also economically, especially due to the use of argon. Argon as a carrier gas is much cheaper than helium and colder than common atmospheric pressure discharges in air such as DBDs, corona or glow discharges.
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