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Impact of selected emulsifiers and different emulsification methods on the growth of Staphylococcus aureus and Escherichia coli.
KOLDOVÁ, Kamila
The main topic of this bachelor's thesis is to determine the minimum inhibitory and bactericidal concentration of selected types of emulsifiers and plant extracts on the growth of Escherichia coli and Staphylococcus aureus bacteria. The thesis also examines the effect of selected emulsifiers and their methods of emulsification on emulsion stability. The theoretical part provides basic information about the bacteria used, their morphological characteristics, pathogenesis, and the diseases they cause. It also describes emulsifiers and their general characteristics, including emulsion stability. The final theoretical part of the thesis focuses on plant extracts, their properties, and their applications. The methodology section of this thesis illustrates the procedures for preparing emulsifiers and plant extracts, as well as determining the minimum inhibitory and bactericidal concentrations. These determinations were performed in a 96-well microtiter plate and measured spectrophotometrically. The method for preparing emulsion stability was conducted in glass test tubes, Eppendorf tubes, and centrifuge tubes. Based on these methodts, the stability of emulsion of Mueller-Hinton broth and sugar solution was determined. The results of minimum inhibitory and bactericidal concentration were processed into tables and evaluated. Emulsion stability was processed and evaluated using obtained photographs.
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USE OF COMPLEX EXTRACTS FROM PLANT AND MICROORGANISMS IN FOODS AND COSMETICS
Uhlířová, Renata ; Kovalčík, Adriána (referee) ; Vorlová, Lenka (referee) ; Márová, Ivana (advisor)
The presented doctoral thesis is focused on optimalization of preparation of extracts of various origin (plant and microbial). The influence of extraction agent, disintegration of microbial cells as well as special techniques for enhancement of extraction process effectiveness (ultrasound and microwaves) were monitored. As part of the characterization, considering the source material, the composition of the prepared extracts was analyzed both spectrophotometrically and using liquid and gas chromatography. The most suitable extraction procedure varies depending on the type of material. For herbal samples, the highest content of active substances was achieved when the extraction facilitated by microwaves was used. Contrarily, the optimal method for cyanobacteria is the traditional extraction method, which is carried out for 24 hours at laboratory temperature. The most suitable method for extracting the active substances from yeast was a combination of DMSO, ethanol and hexane. Further, the effect of extracts on microbial cultures of bacteria (E. coli, S. epidermidis, C. acnes) and yeast (C. glabrata) was observed. For cytotoxicity evaluation MTT and LDH assays were performed on HaCaT and Caco2 human cell lines. The antimicrobial tests have demonstrated the antimicrobial, antibacterial and antifungal effect of a numerous extracts, especially the essential oils of Lavandula angustifolia and Mentha piperita. On the other hand, the cytotoxicity tests showed that the most of the extracts were safe for the use in the food industry and cosmetics within a range of up to 10% by volume. To improve stability, the prepared extracts were enclosed into liposomal particles enriched with PHB polymer (0–100%). Using a ZeteSizer NanoZS device, the particles' size, polydispersity index, and colloidal stability were assessed. The cryoTEM was used to examine the particle´s morphology. Liposomes and liposomes with the addition of 20 % PHB (PHB20) were evaluated as the most suitable particles for encapsulation of the prepared extracts. Excellent stability and high encapsulation efficiency were displayed by the produced particles. Genotoxicity testing with SOS chromotest on the model organism E. coli did not reveal any genotoxic effects. As an alternative method to achieve stabilization of active substances, nanofibers from PHB were prepared by three different techniques and gelatin nanofibers by electrospinning. The fibers were monitored for their diameter, uniformity, distribution on the collector using SEM and the gradual release of active substances from the fiber structure. The evaluations revealed that the forcespinning and electrospinning techniques were the most appropriate for further investigation. A continuous release of active ingredients was achieved by incorporating extracts into nanofibers. Several applications of prepared materials in the food industry and cosmetics were designed and developed as part of this work. The base emulsion's stability and general qualities were improved when green coffee and carotenogenic yeast extracts were added. The usage of emulsions containing coffee and yeast extracts would be suitable as sunscreens, and after the addition of Spirulina extract, as day or night anti-wrinkle creams. Energy drinks of the new type based on coffee with the addition of encapsulated plant extracts enable gradual release of caffeine and antioxidants during digestion.
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