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Adsorption of microbial cells on model biological surfaces
Mikušová, Janka ; Krouská, Jitka (referee) ; Sedláček, Petr (advisor)
The aim of this bachelor thesis is creating sorption surface suitable for studying adsorption bacterial strain Lactobacillus rhamnosus, as representative probiotic bacterial strain. Included in the experimental part of this thesis was the process of creating sorption surface, according to the designed and especially optimized method. Optimization method comprised of preparing sorbent with sorption surface including mucin, on which was the bacterial strain Lactobacillus rhamnosus bound to, using specific interaction. UV-VIS spectrophotometry was utilized in the optimization of the sorption surface preparation for monitoring losses of mucin in solution that happens during the bondation on used substrate. Designed sorption surfaces were subjected to various physical-chemical analyses, which roles were displaying and characterisation of surface and confirming the presence of mucin. One of the major components, in the process of observing and characterization of changes in surface structure of the sorption surface, was infrared spectroscopy with Fourier transformation. Scanning electron microscopy was used for more detailed observation of the surface structure, created by adhesive interactions, which participate in probiotic bacteria adhesion. Finally, the created surfaces were used in pilot experiment, during which, the kinetics in growing bacterial culture with and without the presence of sorbent using microcalorimetry were observed. Designed sorbent appears to be a suitable mediator for detailed characterisation of bacterial adhesion, which is the first step in creating bacterial biofilm.
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Characterization of adsorption properties of probiotic bacteria
Černá, Klára ; Sedláček, Petr (referee) ; Obruča, Stanislav (advisor)
The aim of the diploma thesis is to characterize the adsorption properties of probiotic bacteria. Characterization was performed using of designed and optimized spectrophotometric method of determination of adhesion of probiotic bacteria to mucin as the main viscoelastic component of mucus. In order to provide a more detailed description of the adhesion interactions involved in the adhesion of probiotic bacteria to mucin, the viability of selected probiotic bacterial strains Lactobacillus rhamnosus CCM 1825, Lactobacillus plantarum CCM 7039, Lactobacillus delbrueckii subsp. bulgaricus CCM 7190, Lactobacillus acidophilus CCM 4833, Lactobacillus casei CCM 4798, Bifidobacterium breve CCM 7825, Bacillus coagulans CCM 2658 and a potential probiotic strain of Lactobacillus zeae CCM 7069. Moreover, bacterial adhesion to carbohydrates was also determined for all the bacteria tested. The last proposed and optimized technique was dynamic and electrophoretic light scattering characterizing surface properties such as -potential, average size distribution and isoelectric point. From this method, information was obtained on aggregation of bacterial cells and on electrostatic interactions. The combination of these methods was used as a complex tool to characterize adhesion of the tested bacterial cultures as a very specific, sensitive and key parameter of a successful probiotic microorganism that is influence by the multiple effects.
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Adsorption of microbial cells on model biological surfaces
Mikušová, Janka ; Krouská, Jitka (referee) ; Sedláček, Petr (advisor)
The aim of this bachelor thesis is creating sorption surface suitable for studying adsorption bacterial strain Lactobacillus rhamnosus, as representative probiotic bacterial strain. Included in the experimental part of this thesis was the process of creating sorption surface, according to the designed and especially optimized method. Optimization method comprised of preparing sorbent with sorption surface including mucin, on which was the bacterial strain Lactobacillus rhamnosus bound to, using specific interaction. UV-VIS spectrophotometry was utilized in the optimization of the sorption surface preparation for monitoring losses of mucin in solution that happens during the bondation on used substrate. Designed sorption surfaces were subjected to various physical-chemical analyses, which roles were displaying and characterisation of surface and confirming the presence of mucin. One of the major components, in the process of observing and characterization of changes in surface structure of the sorption surface, was infrared spectroscopy with Fourier transformation. Scanning electron microscopy was used for more detailed observation of the surface structure, created by adhesive interactions, which participate in probiotic bacteria adhesion. Finally, the created surfaces were used in pilot experiment, during which, the kinetics in growing bacterial culture with and without the presence of sorbent using microcalorimetry were observed. Designed sorbent appears to be a suitable mediator for detailed characterisation of bacterial adhesion, which is the first step in creating bacterial biofilm.
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Characterization of adsorption properties of probiotic bacteria
Černá, Klára ; Sedláček, Petr (referee) ; Obruča, Stanislav (advisor)
The aim of the diploma thesis is to characterize the adsorption properties of probiotic bacteria. Characterization was performed using of designed and optimized spectrophotometric method of determination of adhesion of probiotic bacteria to mucin as the main viscoelastic component of mucus. In order to provide a more detailed description of the adhesion interactions involved in the adhesion of probiotic bacteria to mucin, the viability of selected probiotic bacterial strains Lactobacillus rhamnosus CCM 1825, Lactobacillus plantarum CCM 7039, Lactobacillus delbrueckii subsp. bulgaricus CCM 7190, Lactobacillus acidophilus CCM 4833, Lactobacillus casei CCM 4798, Bifidobacterium breve CCM 7825, Bacillus coagulans CCM 2658 and a potential probiotic strain of Lactobacillus zeae CCM 7069. Moreover, bacterial adhesion to carbohydrates was also determined for all the bacteria tested. The last proposed and optimized technique was dynamic and electrophoretic light scattering characterizing surface properties such as -potential, average size distribution and isoelectric point. From this method, information was obtained on aggregation of bacterial cells and on electrostatic interactions. The combination of these methods was used as a complex tool to characterize adhesion of the tested bacterial cultures as a very specific, sensitive and key parameter of a successful probiotic microorganism that is influence by the multiple effects.
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