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Identification of microorganisms in cosmetic products with probiotics
Langová, Denisa ; Španová, Alena (referee) ; Trachtová, Štěpánka (advisor)
Probiotics products are an integral part of the current market. Products containing probiotics cultures are also cosmetic products. The first part of the study focuses on testing of bacterial survival abilities in the environment of preservatives presented in cosmetic products. Collection strains of genus Lactobacillus were used for these tests. Another part of the study focuses on isolation of bacterial DNA from probiotic cosmetic products Ryor, Yoghurt of Bulgaria, FeminaMed and Lactovit Activit in PCR-ready quality. DNA was isolated by fenol extraction and with magnetic particles. Presence of bacteria was proved by genus and species specific PCRs Lactobacillus. Species specific PCR for identification of Lactobacillus pentosus was optimalized. Species identification was in accord with data declared by producers.
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Encapsulation of lactic acid bacteria
Vrtná, Monika ; Lichnová, Andrea (referee) ; Márová, Ivana (advisor)
This bachelor thesis is focused on encapsulation of two strains of probiotic bacteria. Lactic acid bacteria were encapsulated into several types of materials, mostly polysaccharides. The theoretical part describes the techniques of encapsulation applicable in the food industry. During the experimental part alginate and chitosan microparticles were prepared. These particles exhibited different sizes and some of them were modified by starch and carboxymethyl cellulose. Particles were used for encapsulation of two strains - Bifidobacterium breve and Lactobacillus acidophilus. Using flow cytometry cell viability was measured after 24-hours cultivation of bacteria. Particles were observed immediately after encapsulation by optical microscopy and then long-term stability in model foods was evaluated after two days, a week and after four weeks of incubation. Using the Bürker chamber the alived and death bacteria were counted inside and outside the capsule. The stability and viability of the cells were studied also in the artificial intestinal, stomach and bile juices. As the best material for encapsulation of lactic acid bacteria 2% alginate modified by carboxymethyl cellulose was found. The highest viability of bacteria was observed in milk as the real model food. Prepared particles are suitable for use in the food industry.
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Use of high resolution melting analysis for the study of lactic acid bacteria
Knápková, Monika ; Němcová, Andrea (referee) ; Brázda, Václav (advisor)
Currently, there is a growing interest in the use of probiotic products, and there are many of them in the market. With the growing interest, greater emphasis is placed on the identification of declared probiotic microorganisms. Precise identification of microbial composition is often a difficult task and it requires more advanced methods especially in the field of molecular diagnostics. The diploma thesis was focused on the verification of the presence od declared probiotic microorganisms in probiotic food supplements GS Laktobacily Forte 21, Biopron 9 Premium and Linex® Forte. DNA was isolated from the complex matrices by phenol extraction, commercial kit and magnetic carriers F79/L3-PLL in the quality suitable for PCR. Subsequently, the isolated DNA was amplified by real-time polymerase chain reaction using genus- and species-specific primers. The specific PCR product was subjected to agarose gel electrophoresis, whereas species identification was not always in compliance with the data declared by producers. The next part of the thesis was focused on polymerase chain reaction with high-resolution melting analysis to distinguish bacterial strains belonging to the Lactobacillus group and to identify probiotic microorganisms present in the complex matrices of the probiotic food supplements. Eight primer sets were tested (V1F HRM a V1R-HRM, CHAU-V3F a CHAU-V3R, CHAU-V6F a CHAU-V6R, LAC2 a LAC4, LAC1 a LAC2, P1V1 a P2V1, poxcDNAFw a poxPromRVC, poxcDNAFw a poxPromRVT). Three primer pairs (V1F HRM a V1R-HRM, poxcDNAFw a poxPromRVC, poxcDNAFw a poxPromRVT) were evaluated as the most suitable for distinguishing Lactobacillus bacterial strains.
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Current knowledge about the influence of the intestinal microbiome on the development, course and consequences of lactose intolerance
Černá, Pavlína ; Tomešová, Jitka (advisor) ; Kohout, Pavel (referee)
The bachelor thesis investigates the current knowledge about the influence of the gut microbiome on the development and severity of lactose intolerance, as well as whether and how it can be changed to influence the course of existing lactose intolerance and its immediate and long- term consequences. In the first part of this bachelor thesis I discuss lactose itself and its metabolism, which shows its direct link to the gastrointestinal tract. Lactose intolerance (LI) is also described, how it is diagnosed, what it manifests itself as, and what its degrees are. The next chapter discusses the relationship between LI, the gut, and the gut microbiome. First, I focus here on the possible adaptability of the gut, which is mainly determined by the presence of the microbiome. There is also a link between LI and other gastrointestinal disorders in which gut bacteria play a role. The microbiome is a very dynamic population of different types of bacteria, fungi and viruses that can influence the course of lactose intolerance. This can be exploated in the subsequent therapy of LI with probiotics, which are included in part three. The most commonly used probiotic bacteria are representatives of the genus Bifidobacterium and Lactobacillus. However, probiotics are not the only option for using the gut microbiome...
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Influence of experimental intestinal dysbiosis on behavioral, neuroendocrine and immune properties of mice and the possibility of modifying it by administration of probiotic strain Escherichia coli O83:K24:H31
Avramová, Pavla ; Černý, Viktor (advisor) ; Roubalová, Radka (referee)
The gut microbiota is a key factor influencing the function of many organs in the human body, including the nervous, immune, and endocrine systems. This dynamic ecosystem can be modulated by diet, age, environment, even medication. The balanced composition of the microbiota is crucial for individual health, and its disruption leads to dysbiosis. The connection between the gut and the central nervous system is mediated by the microbiota-gut- brain axis. This axis consists of several pathways interconnected to mutually regulate their functions. Among the major components of the axis are the neural pathway, led by the vagus nerve, the immune pathway, and the endocrine pathway, including the HPA axis. Recent studies suggest that probiotic bacteria have the ability to partially rectify dysbiosis and alleviate its consequences for the organism. This thesis focuses on researching the impact of gastrointestinal tract microbiota on the behaviour, immunity, and neuroendocrine aspects of mice. It delves into experimentally inducing dysbiosis using antibiotics and subsequently addressing it with the probiotic bacterium Escherichia coli O83:K24:H31. The aim is to determine whether these probiotics can mitigate the adverse effects of dysbiosis on the immune and neuroendocrine systems, as well as behavioural...
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Effect of early postnatal supplementation by probiotic bacteria Escherichia coli O83:K24:H31 on proportional and fucntional characteristics of selected cellular population
Věcek, Jan ; Hrdý, Jiří (advisor) ; Funda, David (referee)
The hygiene hypothesis proposes that exposure to microorganisms during the postnatal period is crucial for proper immune system development and may help to prevent development of autoimmune diseases and allergies. Probiotics, live microorganisms with beneficial health effects, could be a safe way to promote the appropriate maturation of the immune system. Early postnatal administration of a specific probiotic strain, Escherichia coli O83:K24:H31 (EcO83), reduces the incidence of allergies later in life. To understand the immunomodulatory features of EcO83, we conducted a bioinformatic analysis of its genome and compared it to two other strains, E. coli Nissle and E. coli K12. Our analysis identified unique genes in EcO83 related to propionate and galactose metabolism, as well as genes that may enhance its ability to thrive in the gastrointestinal tract. Moreover, we transformed EcO83 with luciferase enzymes and observed that it effectively colonizes the gastrointestinal tract of newborn mice but not adult mice. Further analysis of mice treated with EcO83 revealed that the probiotic promotes the expression of genes involved in tight junction formation and increases costimulatory molecules on dendritic cells in the mesenteric lymph nodes (MLN). Induced RORγt+ Tregs in MLN displayed increased...
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Interspecific and intraspecific variation in gastrointestinal microbiota composition of parrots and its association with incidence of selected disorders
Černá, Kateřina ; Vinkler, Michal (advisor) ; Rychlík, Ivan (referee)
Captive parrots are susceptible to various digestive, metabolic, and behavioural disorders. Recent research in other vertebrates has suggested that these disorders can be linked to the gut microbiota, directly or through the microbiota-gut-brain axis. Although many commercial probiotic products intended for the use in parrots advertise beneficial effects on digestion and mental well- being, these statements are not sufficiently supported with publicly available scientific evidence, but probably rather based on the extrapolation of the knowledge of microbiota-gut-brain interactions in poultry, mice, and humans. However, there are substantial differences in the gastrointestinal tract morphology and gut microbiota composition between parrots and these model vertebrates. This thesis aimed to describe the interspecific and intraspecific variation in the gut microbiota composition of parrots and to link the variation in microbial communities to the incidence of eight selected behavioural and physiological disorders. The gut microbiota composition of 491 individuals from 85 parrot species was analysed using 16S rRNA metabarcoding. The host species, type of feed, and housing environment were identified as the main factors influencing the diversity and composition of the gut microbiota of parrots. A...
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Alternative beverages of probiotic character
Moravec, Štěpán ; Langová, Denisa (referee) ; Trachtová, Štěpánka (advisor)
Bachelor thesis deals with explanation of therm probiotics, properties of each probiotic bacteria genuses. preparation of coffee and tea, and also deals with probiotic coffee and tea and molekular diagostic methods which were used during bachelor thesis. There were used mikrobiological and molecular diagnostic methods for analysis of probiotic coffee and teas. Namely for confirmation of bacterial genus Bacillus and for confirmation of survavibility of Bacillus during high temperatures. Presence of bacterial DNA in samples of coffee and teas was confirmed by conventional PCR method and by real-time PCR method. Survibility of genus Bacillus during high temperaturewas confirmed by PMA-PCR method.
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Paraprobiotics, postbiotics and other -biotics
Crha, Richard ; Šálek, Petr (referee) ; Trachtová, Štěpánka (advisor)
The theoretical part of the thesis deals with a wide range of microorganisms and substances derived from probiotics. Based on recent publications, probiotics, prebiotics, synbiotics, paraprobiotics, postbiotics, pharmabiotics, psychobiotics and oncobiotics are defined. Furthermore, the purpose and use of the mentioned -biotics is specified for the current understanding of their function. In the experimental part, DNA was isolated from a sample of probiotic capsules using a commercially available bacterial kit and magnetic particles. Its purity and concentration were then determined spectrophotometrically. Using the isolated DNA as a matrix for conventional PCR and detection on agarose gel, the composition of the probiotic product as declared by the manufacturer was demonstrated. Finally, the PMA-PCR method was optimized.
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Identification of probiotics in cometic product using PCR technique
Akmalova, Adelina ; Langová, Denisa (referee) ; Smetana, Jan (advisor)
This bachelor thesis deals with the identification of probiotic microorganisms in cosmetic products by polymerase chain reaction (PCR). The aim of this work was to confirm the presence of bacterial DNA in cosmetic products containing probiotic microorganisms. The theoretical part of the thesis focuses on the role of probiotic microorganisms in maintaining skin health and their effects on the human body. Different methods for the identification of probiotics are also highlighted with a focus on molecular genetic approaches. In the experimental part of the work, two cosmetic products with declared presence of probiotic microorganisms were tested. Two methods were used to extract DNA from the products: phenol extraction and commercial kit method. Furthermore, amplification of a specific region of the 16S rRNA gene was performed by PCR. The PCR products were subjected to agarose gel electrophoresis and visualized by ultraviolet light. All cosmetic products showed the presence of the Bacteria domain and the genus Lactobacillus. In addition, species-specific PCR confirmed the presence of the declared probiotic species Lactobacillus pentosus and Lactobacillus bulgaricus in the products.
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