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Photodynamic inactivation of microorganisms using (aza)phthalocyanine photosensitisers
Holmanová, Pavlína ; Macháček, Miloslav (advisor) ; Kollár, Jan (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Author: Bc. Pavlína Holmanová Supervisor: RNDr. Miloslav Macháček, Ph.D. Consultant: RNDr. Klára Konečná, Ph.D. Title of diploma thesis: Photodynamic inactivation of microorganisms using (aza)phthalocyanine photosensitisers Background: This work is aimed to optimize methodological approaches for the evaluation of the antimicrobial activity of selected representative (aza)phthalocyanine photosensitizers (PSs) in vitro. In addition, the in vitro antimicrobial activity of candidate PSs was evaluated against three clinically important pathogens. Namely, gram positive methicillin resistant bacterium Staphylococcus aureus (MRSA), gram negative bacterium Pseudomonas aeruginosa (PA), and yeast Candida albicans (CA) were included in the study. Methods: Microorganisms were incubated with the selected PSs, which were subsequently washed away, and microorganisms with bound PSs were irradiated for 15 or 30 min. At the same time, the intrinsic antimicrobial activities of the studied compounds without irradiation were determined as well. Three different methodological approaches were used to evaluate the minimum bactericidal concentration (MBC)/minimum fungicidal concentration (MFC). The Bioscreen C instrument was used for...
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Photodynamic inactivation of microorganisms using (aza)phthalocyanine photosensitisers
Holmanová, Pavlína ; Macháček, Miloslav (advisor) ; Kollár, Jan (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Author: Bc. Pavlína Holmanová Supervisor: RNDr. Miloslav Macháček, Ph.D. Consultant: RNDr. Klára Konečná, Ph.D. Title of diploma thesis: Photodynamic inactivation of microorganisms using (aza)phthalocyanine photosensitisers Background: This work is aimed to optimize methodological approaches for the evaluation of the antimicrobial activity of selected representative (aza)phthalocyanine photosensitizers (PSs) in vitro. In addition, the in vitro antimicrobial activity of candidate PSs was evaluated against three clinically important pathogens. Namely, gram positive methicillin resistant bacterium Staphylococcus aureus (MRSA), gram negative bacterium Pseudomonas aeruginosa (PA), and yeast Candida albicans (CA) were included in the study. Methods: Microorganisms were incubated with the selected PSs, which were subsequently washed away, and microorganisms with bound PSs were irradiated for 15 or 30 min. At the same time, the intrinsic antimicrobial activities of the studied compounds without irradiation were determined as well. Three different methodological approaches were used to evaluate the minimum bactericidal concentration (MBC)/minimum fungicidal concentration (MFC). The Bioscreen C instrument was used for...
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Overview of methodological approaches leading to qualitative anad quantitative microbial biofilm evaluation
Holmanová, Pavlína ; Konečná, Klára (advisor) ; Vejsová, Marcela (referee)
Charles Univerzity Faculty of Pharmacy in Hradec Králové Study program: Medical Laboratory Technician Author: Pavlína Holmanová Supervisor: RNDr. Klára Konečná, Ph.D. Title of bachelor thesis: Overview of methodological approaches leading to qualitative and quantitative microbial biofilm evaluation Background: The aim of this work was to compose an overview of methodical approaches leading to qualitative and quantitative evaluation of microbial biofilms. To optimize the methods leading to preparation of mounts with biofilm-forming microorganisms evaluated by epifluorescence or confocal laser scanning microscopy. Methods: Optimization of microbial biofilm staining procedures for subsequent evaluation by epifluorescence microscopy. Formation of microbial biofilms - cultivation of microorganisms in various cultivation media in a microtiter plate, preparation of solutions with fluorescent dyes. Observation of coloured, dispersed biofilms by fluorescence microscope. Results: Using fluorescence microscopy, optimal target concentrations of fluorescent dyes in various combinations were verified. The combination of Syto 9 (SYTO™ 9 Green Fluorescent Nucleic Acid Stain) and propidium iodide was confirmed to be effective for the determination of viability of biofilm-forming microbial cells. The combination of...
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