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DNA isolation from probiotic lactic acid bacteria in food additives
Tvrdíková, Jana ; Vojtíšková, Marie (referee) ; Španová, Alena (advisor)
In this work the functionalised magnetic particles were tested with streptavidin to selective DNA isolation. The method of selective DNA isolation was tested by using DNA probiotic strain Lactobacillus paracasei subsp. paracasei CCDM 211/06. A test was done on the biotinyl oligonucleotic particles, which was immobilised by containing streptavidin and it was used like a DNA probe for isolation complementary DNA chain by means of DNA/DNA hybridization. The primer R 5´ bio and the biotinyl denatured specific PCR product were tested for species Lb. paracasei as a DNA probe. These following experimental conditions were optimized for selective DNA isolation: temperature and time of hybridization, amount of DNA and the release of DNA from microspheres. Isolation of DNA was verified by PCR with specific generic primers. The specific generic PCR product was amplified in extent 250 bp, which was detected by using electrophoresis in agarose gel. This optimized method was successfully used in selective isolation of DNA Lactobacillus from a complementary sample of supplementary food (BIFI pangamin).
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Comparison of microbial metabolic production from waste and powder whey
Elefantová, Petra ; Vítová, Eva (referee) ; Babák, Libor (advisor)
The master’s thesis discusses the comparison of microbial metabolic production from waste and powder whey. Whey is obtained as a by-product of cheese production. Lactose (preferably whey) using lactic acid bacteria (eg. Lactobacillus) under suitable temperature conditions is converted to lactic acid. Effect of temperature, effect of salts and effect of yeast extract on lactic acid production by L. casei were investigated. HPLC metod was determined lactid acid. In the practical part were used bacteria of the genus Lactobacillus. It was found that for dried and waste whey is the optimal temperature of 35 °C. At this temperature is the greatest gain of lactic acid. The highest concentration of lactic acid was obtained by using 20 g of yeast extract for dried whey and for waste whey were used 24 g of yeast extract. When monitoring the effect of salt concentration on the production of lactic acid, it was found that using only MnSO4·H2O gain most of lactic acid.
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Lactic acid bacteria and their utilization
Jiříková, Ivana ; Vránová, Dana (referee) ; Babák, Libor (advisor)
This bachelor thesis is aimed at the study of lactic acid bacteria. Within the frame of this thesis I have performed a literature research into findings about these bacteria. The first section is focused on the general bacteria identification and division into separate genus groups. It also describes the cytology, metabolism and industrial use of milk fermentation bacteria.
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Importance of probiotics in child's nutrition
Augustová, Kamila ; Vítová, Eva (referee) ; Vránová, Dana (advisor)
With great advances in science, medicine and knowledge of a man at the high level of treatment. However, even today the humanity seeking to return to its roots - the nature and use it to maintain its good health. The term probiotics is meant by living organisms that are capable of resettling in the human intestine and have a positive impact on maintaining healthy intestinal microorganisms, which also reflected the positive effects on human health in general. Company Valosun a.s. provided for testing probiotický food supplement for children, which was tested mothers with children preschool age and pregnant. The aim of our sociological survey was to determine what is the awareness of the issue of probiotics in this group, the perception of the queried the availability of probiotics and what are the most common reasons for their use.
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Testing of lactic acid bacteria Lactobacillus genus for bacteriocins production
Volecová, Veronika ; Němcová, Andrea (referee) ; Trachtová, Štěpánka (advisor)
Antimicrobial substances, or bacteriocins are substances produced by probiotic lactic acid bacteria. They have a positive effect on the gastrointestinal tract of humans and are especially suitable for the food, but also the pharmaceutical industry. The aim of the thesis was the molecular identification of lactic acid bacteria of the Lactobacillus genus, species and their subsequent inclusion PCR method. Using the PCR method were tested also genes responsible for the production of bacteriocins. To confirm the production of bacteriocins has been selected the microbiological method, agarose droplet spot-test. In the present study also included the bioinformatics part to assess the specificity and non-specificity of the primers using in Primer-BLAST program.
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Selected bioengineering characteristics of lactic acid bacteria
Šťásková, Lucie ; Vítová, Eva (referee) ; Babák, Libor (advisor)
The diploma thesis deals with the growth of biomass and production of selected metabolit–lactic acid by thermophilic bacteria Bacillus coagulans. The resulting selected metabolite was determined by HPLC method. Cultivations of this genus were performed on synthetic media, where the influence of carbohydrate used as carbon source was tested. Lactose was more suitable fot growth of biomass and glucose for production of lactic acid. On natural whey media the influence of different conditions were tested. The highest yields of biomass and production of lactic acid were observed on enriched whey medium. The last part deals with comparing the production of biomass and metabolites, depending on the volume of media. There were compared selected bioengineering characteristics of all cultivations.
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Molecular typisation of lactic acid bacteria
Jelínek, Zdeněk ; Ing. Špano Miroslav,PhD. (referee) ; Rittich, Bohuslav (advisor)
Lactic acid bacteria play important role both in food industry and health care. They are part of many fermented dairy products and gastrointestinal microflora. There is a whole variety of methods for bacteria identification, but we will focus on Rep-PCR and RAPD only. The aim of this work will be to discuss the impact of different correlation coefficients and cluster analysis algorithms on the results of taxonomic analysis.
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The analysis of DNA isolated from different types of probiotic products using real-time PCR and HRM analysis
Sedláková, Lucie ; Rittich, Bohuslav (referee) ; Trachtová, Štěpánka (advisor)
The aim of this diploma thesis was to introduce real-time PCR with high-resolution melting analysis for Bifidobacterium species. Currently a small number of publication, dealing with identification of Bifidobacterium species using high-resolution melting analysis, is available. According to publications dealing with identification of lactic acid bacteria were selected primers P1V1 and P2V1, LAC1 and LAC2, LsppUPF and LsppUPR, V3F and V3R, V6F and V6R. Using this primers bacterial DNA was amplified by real-time PCR with high-resolution melting analysis. After evaluation of the measured results efficiency of selected primers was verified on DNA izolated from complex sample of probiotic product. After further optimisation real-time PCR with high-resolution melting analysis could be suitable using selected primers for Bifidobacterium species.
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Identification of lactic acid bacteria in probiotic products (tablets) using amplification methods
Balogová, Petra ; Trachtová, Štěpánka (referee) ; Španová, Alena (advisor)
Lactobacillus have been playing a crucial role in the fermentation processes for millenia. Their probiotic effects have been studied deeply and nowadays they are used in the production of fermented products and food adjunkt including probiotic preparations. This thesis was focused on bakteria of genus Lactobacillus in probiotic preparations (tablets). Polymerase chain reaction method (PCR) based on amplification of DNA was used to identify lactobacillus in seven probiotic preparations. As DNA matrices whole DNA recovered from tablets with the help of magnetic microparticles (P(HEMA-co¬-GMA) coated by carboxylic groups was used into a PCR mixture. Genus specific PCR products (250 bp) were detected with the help of gel electrophoresis on agaróse. DNA of genus Lactobacillus was proved in all but one product. In this product only probiotic bacteria were declared presence without family identification was proved. It contained different bakteria then bakteria of Genus Lactobacillus probably.
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Methods for detection of antimicrobial properties of lactic acid bacteria
Vránová, Petra ; Němcová, Andrea (referee) ; Trachtová, Štěpánka (advisor)
Lactic acid bacteria are classified as probiotics producing substances that act against microorganisms. These antimicrobial substances include organic acids, carbon dioxide, hydrogen peroxide and bacteriocins. Currently, the focus is on bacteriocins, which are used in the food and pharmaceutical industries. The aim of this thesis is creating an overview of antimicrobial properties of lactic acid bacteria, methods of their determination and their applications in the food industry. The experimental part deals with identification of lactic acid bacteria present in selected samples of bacterial DNA. In addition, we tested for bacteriocin Gassericin K7A-producing gene. Molecular diagnostic methods such as real-time polymerase chain reaction (qPCR) and conventional polymerase chain reaction were used to distinguish bacterial DNA as well as the bacteriocin.
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