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Effect of dysbiosis on proportion of particular neutrophil subsets and their functional capacities
Sklenářová, Lydie ; Hrdý, Jiří (advisor) ; Dobeš, Jan (referee)
The gut microbiota is crucial for maintaining physiological balance and influences metabolic processes, immune responses, and intestinal barrier function. Dysbiosis, or the imbalance of microbial composition, is associated with a range of health complications, including chronic inflammatory conditions such as non-specific intestinal inflammations. Inflammatory processes associated with dysbiosis and changes in microbial metabolites can directly affect the activation of neutrophils, impacting the pathogenesis of various diseases. Probiotics, defined as live microorganisms which, when administered in adequate amounts, confer a health benefit on the host, offer the potential for positive modulation of these inflammatory conditions The aim of this thesis was to explore how experimentally induced intestinal dysbiosis affects the heterogeneity of neutrophils in the bone marrow. Dysbiosis was induced by administering antibiotics to mice, which were subsequently treated with the probiotic strain Escherichia coli O83:K24:H31 (EcO83). Neutrophil phenotypes were assessed using flow cytometry based on the expression of surface markers CD11b, Ly6G, CD62L, and CXCR2. Meanwhile, gene expression related to their antimicrobial functions and the inflammatory environment was analyzed by quantitative PCR. The results...
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The potencial of next-generation probiotics
Kharkevich, Viktoryia ; Langová, Denisa (referee) ; Trachtová, Štěpánka (advisor)
The aim of the bachelor thesis was to examine the possibilities of identifying probiotic microorganisms using molecular diagnostic techniques such as PCR and HRM-PCR in a commercially available product – Probiolact Forte 12. In the theoretical part, a literature review focused on next-generation probiotics was conducted. In the practical part, cultivation of microorganisms contained in one capsule in liquid medium and solid media was performed. DNA isolation was carried out using the NucleoSpin Microbial DNA commercial kit according to the manufacturer's instructions. The purity and concentration of isolated DNA were verified using spectrophotometric analysis. Primers for specific PCR analysis of DNA domains Bacteria and individual genera and species, whose presence the manufacturer declares in the preparation, were used for DNA identification. The results of the PCR analysis confirmed the presence of the target DNA, i.e., bacterial genera and species in the preparation, as claimed by the manufacturer.
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Development of active supplement with probiotics
Cveček, Jan ; Dzurická, Lucia (referee) ; Márová, Ivana (advisor)
Subject of this bachelor thesis was to investigate the antimicrobial activity of selected probiotic strains and determine how the probiotic growth is affected probiotic growth by prebiotics and essential oils. Probiotics are bacteria, which positively affect human health. They are mainly used in food industry as fermentation starter cultures, where they produce antimicrobial compounds that support resistance to pathogenetic bacteria during the fermentation process. In general, fermentation process is used to prolong the shelf-life of foodstuff. However, the applications of probiotics are very limited in food or other industry because probiotics are living organisms, so at least the industry tries to use their metabolic products. The method of encapsulation could support the usage of probiotics in other unusual ways. In experimental part of this thesis, the growth of probiotics was studied in both commercial MRS broth and in minimal broth, which could help with isolation of bioactive metabolic products such as lactic acid and bacteriocins. Additionally, the effect of prebiotic-inulin and essential oils from oregano and thyme on probiotics growth was investigated. In the second part of thesis, the content of lactic acid was analysed and antimicrobial activity of selected probiotic strains and effect of additives on activity was tested. Antimicrobial test discovered that prebiotics and essential oil from thyme enhanced the antimicrobial activity of tested samples. Also, the minimal broth was shown to be unsustainable for cultivation of used probiotics because selected strains exhibited small production of biomass and very low antimicrobial activity. In the third part, the probiotic strain was encapsulated with prebiotic and essential oil into alginate particles. Bacterial growth from the particles in MRS broth was investigated over period of 15 days. Results of this measurement were that probiotics were gradually released from particles over the period. Subsequently the potential use of these particles was proposed for further investigation.
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Influence of some medicinal fungi and other prebiotics on growth and metabolism of probiotic bacteria
Husová, Martina ; Bendová, Agáta (referee) ; Márová, Ivana (advisor)
The bachelor thesis deals with the influence of medicinal mushrooms and other prebiotics on the growth and biological activity of probiotic bacteria. Selected genera of lactic acid bacteria Bifidobacterium and Lactobacillus can produce antimicrobial substances bacteriocins and their production depends on cultivation conditions and available nutrient sources. The theoretical review describes and characterizes probiotic bacteria, their metabolism and importance for human health. Similarly, the theoretical section focuses on prebiotics and their selected representatives, which are four species of medicinal fungi, the microscopic algae Chlorella, resistant starch, the cell biomass of the yeast Saccharomyces cerevisiae and dietary supplements such as inulin and psyllium. In the experimental part, extracts were prepared from these prebiotics by three different methods - boiling, macerating and hydrolysis. The obtained extracts were characterized in terms of macronutrient content such as carbohydrates and proteins. Based on the growth of four species of lactic acid bacteria cultured in standard medium and their production of antimicrobial substances, a strain of B. bifidum was selected for cultivation in the presence of prebiotic extracts. After determining the increase in probiotic biomass in the prebiotic extracts, the cultures were centrifuged, and the protein content of the bacterial supernatant was determined. The lactic acid content of the lyophilized probiotic extracts was determined by high-performance liquid chromatography, and finally, the antimicrobial activity was determined by broth dilution method and resazurin viability assay. The results of the antimicrobial tests were compared with the antimicrobial activity of lactic acid at the same concentration as that contained in the probiotic extracts. For the highest macronutrient content of the prebiotic extracts, extraction by boiling was the most suitable method. For insoluble polysaccharides such as resistant starch, hydrolysis might be a better option, but this requires subsequent treatment of the extract to provide suitable conditions for cultivation. The boiled extracts were the best for the growth of probiotic bacteria as well as their production of antimicrobial substances. The highest concentrations of protein and lactic acid were measured in these samples. The likely presence of bacteriocins was confirmed in all culture types. Almost all probiotic extracts showed antimicrobial activity against Staphylococcus epidermidis and Escherichia coli. The likely presence of bacteriocins was confirmed by the broth dilution method, for example in extracts containing chlorella algae or S. cerevisiae yeast, because these extracts were antimicrobial at lower concentrations than the lactic acid present.
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Evaluation of the presence of probiotic bacteria in a food supplement using molecular biological methods
Dvornyi, Nikolai ; Brázda, Václav (referee) ; Smetana, Jan (advisor)
Probiotics are beneficial microorganisms that, when administered properly, can provide health benefits to the host, such as aiding digestion and boosting the immune system. Their effectiveness and benefits depend on the specific strain of microorganism. These strains are often from the genera Lactobacillus, Bifidobacterium, and represent natural hosts of the gastrointestinal tract. This bachelor thesis focuses on the identification of probiotic bacteria in a commercially available dietary supplement Linex® Forte using molecular biological methods, mainly polymerase chain reaction (PCR). The aim was to verify the presence of the claimed probiotic cultures of Lactobacillus acidophilus, Bifidobacterium animalis, and to compare the results with the manufacturer's claims. Therefore, two DNA isolation methods were used: phenolchloroform extraction and the commercial purification kit OMNI International. The isolated DNA was then analyzed by domain, genus and species-specific PCR. The results show that the methods used are suitable for the identification of probiotic bacteria and confirm their presence in the selected product.
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Utilization of PCR technique for identification of probiotic bacteria in daily hygiene product
Horobets, Yuliia ; Fialová, Lenka (referee) ; Smetana, Jan (advisor)
Probiotic bacteria have traditionally been used in the food industry, but their applications have now expanded to the prevention and treatment of various diseases. Increasing evidence supports the efficacy of bacteria of the genus Lactobacillus in the oral cavity has led to the application of probiotic strains of this genus in mouthwashes and other personal care products. In this bachelor thesis, DNA was isolated by two methods, then quantified spectrophotometrically and amplified via conventional PCR. The results of the polymerase chain reaction were detected by gel electrophoresis. The presence of probiotic bacteria species Lactobacillus delbrueckii, Lactobacillus plantarum and Lactobacillus pentosus was confirmed.
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Verification of the presence of probiotic bacteria in a cosmetic product using the polymerase chain reaction (PCR) technique
Raevskaya, Vera ; Kroupová, Zuzana (referee) ; Smetana, Jan (advisor)
This bachelor thesis focuses on the verification of the presence of probiotic bacteria in a cosmetic product using the polymerase chain reaction (PCR) technique. The theoretical part provides an overview of probiotics, their application in cosmetics and their effect on skin condition. Furthermore, the most commonly used molecular biological methods for the identification of probiotic organisms are described. In the practical part, the work focuses on the isolation of bacterial DNA from cosmetic products and its subsequent amplification by PCR. The amplified products were analyzed by agarose gel electrophoresis and detected by UV irradiation. The results showed the presence of bacteria of the genera Lactobacillus and Bifidobacterium, but the presence of bacteria of the genus Lactococcus, which were declared by the manufacturer together with the above genera, was not confirmed.
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Použití alternativních aditivních látek v odchovu telat
VALTOŠOVÁ, Gabriela
The diploma thesis consists of a literature research, which is then used for compari-son with the results of the experiment and a practical part investigating the effect of various prebiotics and probiotic preparations on the health and growth of calves in the period of colostrum and milk nutrition.
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Optimization of Hericium erinaceus Cultivation and Subsequent Fermentation with Probiotics
Radvanová, Tereza ; Vršanská, Martina (referee) ; Hudečková, Helena (advisor)
The focus of this diploma thesis is to optimize cultivation of the hericium erinaceus mushroom on cereal substrates with subsequent fermentation with probiotic bacteria. Theoretical part of this thesis focuses on description of the mushroom hericium erinaceus and its nutritional value, biological properties, other uses and cultivation. Furthermore, an overview of the nutritional value of the used cereal substrates is provided, along with the characterization of probiotics, their biological effects, and metabolism. Finally, a chapter dedicated to the description of the traditional koji fermentation process was included. In the experimental part, the cultivation of hericium erinaceus was optimized on seven cereal substrates – barley groats, cornmeal, wheat bran, buckwheat, amaranth, white and red quinoa. Using spectrophotometric methods, the content of water soluble proteins, total carbohydrates, total phenolic compounds, flavonoids, and antioxidant activity was determined in the cereal substrates themselves and substrates enriched with mycelium. Furthermore, the content of fatty acids was determined in the samples using the gass chromatography method and lipophilic compounds were analyzed using high performance liquid chromatography. Cereal substrates enriched with mycelium showed increased antioxidant activity and concentrations of water soluble proteins, phenolic compounds, flavonoids, and lipids. Moreover, elevated concentrations of tocopherols, ubiquinone, and ergosterol were detected through high performance liquid chromatography analysis. Substrates with abundant mycelial growth, such as barley groats, cornmeal, red quinoa, and amaranth, were selected for fermentation with probiotic bacteria Lactobacillus rhamnosus. After the fermentation process, spectrophotometric methods were repeated, and the concentration of lactic acid was determined using high performance liquid chromatography. Samples with probiotics showed a slight increase in their antioxidant activity, concentration of phenolic compounds, flavonoids, and water soluble proteins. The concentration of lactic acid in control samples and samples with probiotics was almost unchanged. The obtained results from this study suggest that cereals enriched with mycelium and the fermentation product of probiotics could be utilized in the development of dietary supplements or functional foods.
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