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Detection of probiotic bacteria in diary food products using PCR technique
Klaška, Dominik ; Brázda, Václav (referee) ; Smetana, Jan (advisor)
In the bachelor thesis, DNA was isolated from commercially available white yogurt. The isolated DNA, which was gained by two different methods, was performer analyse by a spectrophotometer. Both methods provided sufficiently concentrated and high-quality DNA for further analysis by PCR. Precisely defined sections of isolated DNA were amplified using specific primers. The presence of the bacterium domain was detected, and in the case of genus specific amplification, the presence of bacteria of the genus Lactobacillus was detected too, by gel electrophoresis.
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Influence of some foods on growth and viability of probotic bacteria
Vajglová, Klára ; Němcová, Andrea (referee) ; Márová, Ivana (advisor)
The goal of this work was a study the influence of food and beverages on the viability and growth of probiotic bacteria. The influence of food and beverages was tested on monocultures of Lactobacillus acidophilus, Bifidobacterium breve and mixed culture of probiotic microorganisms. In the experimental part, probiotic cultures were incubated in selected foods and beverages. After that they were tested in a model conditions of digestive tract. In some probiotic cultures, growth of viable cells during incubation in the digestive tract was observed. The increase of probiotic cells was showed predominantly in foods that contained higher levels of sugars and fats or a suitable combination. Their increase was up to four times in some cases. Based on the results, mixed probiotic cultures aren’t surprisingly exhibited better survival and maintain sufficient amount of viable cells even during the digestive process. Moreover, probiotic microorganisms could be recommended to consumption during meals better than just with a beverage.
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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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Preparation of food supplements for children containing probiotic bacteria and fruit components
Vetchá, Vendula ; Bendová, Agáta (referee) ; Skoumalová, Petra (advisor)
This bachelor thesis is focused on the preparation of supplementary food with probiotic culture and fruit components. The aim of this work was to determine which of the selected extracts will affect the growth of probiotics and also will contain a certain proportion of phenolic compounds, flavonoids, vitamin C and show antioxidant effects. The theoretical part of the work is focused on the intestinal microbiota, the issue of probiotics and the characterization of encapsulation methods of probiotics. We used extracts of teas and fruit syrups in the experimental part. The antimicrobial activity against model microorganisms Escherichia coli, Micrococcus luteus, Staphylococcus epidermidis and Serratia marcescens was analyzed. Interaction of probiotic bacteria Lactobacillus acidophilus, Lactobacillus casei and Bifidobacterium breve in extracts were tested. After model digestion, cell viability of probiotic bacteria in combination with extracts was examined during and after digestion. In the last part, encapsulated particles containing probiotics and a mixture of tea and syrup were prepared.
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Viability of probiotic bacteria in some foods
Pokorná, Martina ; Bokrová, Jitka (referee) ; Márová, Ivana (advisor)
This bachelor thesis is focused on viability od probiotic bacteria in some foods and looking for optimal food and beverages, that exhibited a minimal negative effect on the growth and viability of probiotic bacteria. Lactobacillus acidophilus, Bifidobacterium breve and commercially complex of probiotics were tested. In the practical part, probiotics were incubated in the environment of selected types of model and real foods. Further, incubation in model gastrointestinal juices to simulate human digestion was performed. Model foods with a moderate value of concentrations of sugar, protein and alcohol were shown the best alternative for growth of bacteria and their lowest mortality. For salt, the smallest concentration was shown as the most suitable. The chicken broth, chicken and beef meat were evaluated as the best environment for the capsule with complex probiotics. The vegetables and vegetable broth were stated like the best for Lactobacillus acidophilus and Bifidobacterium breve. According to the preliminary results we can conclude that the probiotics should be preferably consumed during meals and with the sufficient amount of liquids for the activation of all cells.
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Using different methods of DNA isolation of lactic acid bacteria in molecular biological methods
Chvalkovská, Eva ; Skoumalová, Petra (referee) ; Brázda, Václav (advisor)
This thesis focused on the probiotic bacteria, DNA isolated from these bacteria by three different methods and the effect of isolation on DNA identification using molecular biological methods. Probiotic bacteria are an important part of human intestinal tract. They have an important role in the function of the immune system due to adhesion to the mucosa of the intestinal flora. They create a inhostile environment for pathogens. Probiotic bacteria are commonly taken in the food like dairy products or food supplements. However, overuse of antibiotics is at risk of passing on the intrinsic resistance that probiotic bacteria have to the pathogenic bacteria. The intrinsic resistence they have to maintain the natural homeostasis of the intestinal tract. It is important to effectively identify risky probiotic bacteria that have the ability to transmit resistance to eliminate their presence in food and dietary supplements. Three methods of DNA isolation like phenol extraction method, magnetic particle isolation and commercial kit isolation were used in the experimental part. DNA was isolated from three dietary supplements, namely Biopron 9 premium, Linex forte and GS Lactobacily forte 21. The purity and concentration of the isolated DNA was detected spectrophotometrically. The presence of individual DNA strains in dietary supplements was confirmed by real-time polymerase chain reaction. The best method of isolation in terms of purity and concentration of isolated DNA was evaluated by RT-PCR and spectrophotometry using a commercial kit isolation method.
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DEVELOPMENT AND TESTING OF NATURAL FOOD COMPOUNDS FOR INFANTS´ NUTRITION
Hoová, Julie ; Kráčmar, Stanislav (referee) ; Vítová, Eva (referee) ; Márová, Ivana (advisor)
The presented doctoral thesis is focused on development and testing of natural compounds for infant’s nutrition. Selected groups of plant-based food were examined from nutritional point of view, as well as regarding the content of biologically active compounds, especially compounds with antioxidant and antibacterial activity. As basic nutrients were measured following groups of compounds: total saccharides, reducing sugars, insoluble fiber, total nitrogen, and fatty acid profile. Moreover, content of specific vitamins and provitamins, total phenolic compounds, flavonoids, antioxidant capacity, natural pigments, fructans, -glucans and organic acids were determined in selected samples. The evaluation of antimicrobial effect was studied via interaction with appropriate types of microbial cells. The safety of tested materials was verified by cytotoxicity tests using human cell lines HaCaT and Caco-2. In next part of present work, encapsulation techniques were applied to enhance stability and consumer acceptance of natural extracts. Materials were encapsulated into nanoparticles, especially into liposomes of different composition. Size distribution, uniformity, zeta potential, encapsulation efficiency and release control were monitored. Sunflower and soya bean lecithin were determined as the most suitable for preparation of stable liposomes. Further part of the study was focused on the role of probiotics in infant´s nutrition. Selected probiotic strains were cultivated and the influence of the encapsulation techniques and presence of plant extracts on probiotic viability were examined in natural form and in model digestion conditions. Cultivation techniques and flow cytometry were performed to evaluate probiotic viability. Based on the results obtained, plant extracts rich in bioactive compounds and having potential effect on human organism and probiotic bacteria were selected. As a most suitable combination, green barley and some probiotics like lactobacilli and bifidobacteria could be recommended. Finally, the specific study focused on the transfer of selenium species from mother to child via colostrum was analyzed. Selenium is a biogenic element acting as a representative of early natural antioxidants in human body. The important role of colostrum in neonatal nutrition was confirmed by the detection of the presence of selenoprotein P in the colostrum. This protein is further taken up by the child immediately after birth and contributes as the earliest antioxidant to infant´s immunity. Overall, it can be concluded that, despite the undeniable fundamental importance of protein in childhood nutrition, further plant-based biologically active compounds and combinations of probiotics and prebiotics, represent a major benefit and contribute positively to the increase of the nutritional value and quality of foods for infant nutrition.
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Preparation of food supplements for children containing probiotic bacteria and fruit components
Vetchá, Vendula ; Bendová, Agáta (referee) ; Skoumalová, Petra (advisor)
This bachelor thesis is focused on the preparation of supplementary food with probiotic culture and fruit components. The aim of this work was to determine which of the selected extracts will affect the growth of probiotics and also will contain a certain proportion of phenolic compounds, flavonoids, vitamin C and show antioxidant effects. The theoretical part of the work is focused on the intestinal microbiota, the issue of probiotics and the characterization of encapsulation methods of probiotics. We used extracts of teas and fruit syrups in the experimental part. The antimicrobial activity against model microorganisms Escherichia coli, Micrococcus luteus, Staphylococcus epidermidis and Serratia marcescens was analyzed. Interaction of probiotic bacteria Lactobacillus acidophilus, Lactobacillus casei and Bifidobacterium breve in extracts were tested. After model digestion, cell viability of probiotic bacteria in combination with extracts was examined during and after digestion. In the last part, encapsulated particles containing probiotics and a mixture of tea and syrup were prepared.
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Detection of probiotic bacteria in diary food products using PCR technique
Klaška, Dominik ; Brázda, Václav (referee) ; Smetana, Jan (advisor)
In the bachelor thesis, DNA was isolated from commercially available white yogurt. The isolated DNA, which was gained by two different methods, was performer analyse by a spectrophotometer. Both methods provided sufficiently concentrated and high-quality DNA for further analysis by PCR. Precisely defined sections of isolated DNA were amplified using specific primers. The presence of the bacterium domain was detected, and in the case of genus specific amplification, the presence of bacteria of the genus Lactobacillus was detected too, by gel electrophoresis.
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