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Biofilm formation in probiotic cultures and its application in pharmacy
Ryšávka, Petr ; Obruča, Stanislav (referee) ; Vorlová, Lenka (referee) ; Márová, Ivana (advisor)
The work was comprehensively focused on the development of adhesive forms of probiotics in the form of a biofilm on combined carriers with a prebiotic component. The second part dealed with the influence of food on the multiplication and survival of selected types of probiotic bacteria. Subsequently, the effect of individualized probiotic supplements on changes in the human intestinal microbiome was monitored. Suitable adherent probiotic strains for biofilm formation were selected and tested. Methods have been introduced and different variants of carriers for culturing and binding bacteria have been tested. In vitro experiments verified the stability of biofilm stucture and its resistance to low pH, bile and antibiotics in comparison with the planktonic cell form. The antimicrobial effect of probiotic strains in the form of a biofilm was studied. The cultivation of the multispecies biofilm on the combined carrier was optimized and the stability of the biofilm and the final viability of probiotic bacteria were confirmed. Furthermore, the influence of various foods and beverages on the viability of probiotic bacteria was evaluated with emphasis on the simulation of passage through the gastrointestinal tract. Both models, solutions with standardised concentrations of alcohol, sugar, salts, proteins or different pH and different types of real foods and beverages were tested. The effect of food and beverages was tested on monocultures of Lactobacillus acidophilus, Bifidobacterium breve and on probiotic capsules containing a mixed culture of probiotic microorganisms. The survival of probiotics in various food matrices in the simulated gastrointestinal tract was quantitatively different. We managed to define foods suitable for supporting the multiplication of probiotic bacteria. A separate part of the work was focused on the targeted modulation of the intestinal microbiome by individualized probiotics that were prepared on the basis of molecular biological analyzes of the intestinal microbiome aimed at detecting the percentage of lactobacilli, bifidobacteria and phylum Firmicutes and Bacteroidetes. Personalized probiotic supplementation confirmed the positive effect of this approach on microbiome changes, especially on the increase of the content of lactobacilli, bifidobacteria and the overall diversity of the microbiome.
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Influence of selected types of food on changes in the composition of the intestinal microbiome
Šantavý, Tomáš ; Němcová, Andrea (referee) ; Ryšávka, Petr (advisor)
The master's thesis focuses on determining the influence of selected types of food on changes in the intestinal microbiome composition. Dietary composition plays an important role in controlling intestinal microbial populations, including the prevention and treatment of certain diseases. In order to evaluate the effect of daily consumption of fruit-vegetable smoothie on the human intestinal microbiome, changes in the intestinal microbiome at the beginning and after 14 days of intervention were compared. The experimental part was attended by 10 men and 10 women aged 35-45 years, who were evenly divided into 2 groups. The first group of participants consumed a combination of orange, dried turmeric and ginger. The second group consumed a combination of apple, carrot and lemon juice. DNA isolation from stool samples followed by a quantitative PCR method was used to detect differences in the intestinal microbiome. After 14 days of consumption of the fruit-vegetable smoothie, specific changes in the phylum Firmicutes, Bacteroidetes and the genus Lactobacillus, Bifidobacterium were observed. After statistical evaluation of the obtained results, the differences were not detectable at a statistically significant level (p-value > 0.05).
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Screening of probiotics bacterial strains aimed at proof of gluten degradation
Korvasová, Lucie ; Obruča, Stanislav (referee) ; Ryšávka, Petr (advisor)
No other treatment for celiac disease and other gluten-related disorders has been found so far except for a lifelong diet, which is significantly restrictive for the patient. Moreover, it is not always possible to adhere strictly to this diet, mainly due to random gluten ingestion, which leads to health complications associated with these diseases. Currently, vaccination is being tested as an alternative treatment, and there is also increasing attention paid to enzymes that could break down the immunogenic part of gluten, specifically in its gliadin component. This thesis examined bacteria that could potentially break down this sequence in gliadin and thereby reduce its concentration during detection. A considerable proportion of the tested bacteria belong to probiotics to ensure their future use for humans is safe. To obtain data, the fact was utilized that as bacteria grow, the turbidity increases proportionally, which was subsequently detected to determine the amount of bacterial growth. The RIDASCREEN® Gliadin competitive kit was used for the gliadin detection test, which works on the principle of a competitive ELISA method with R5 antibodies. The result of the study was the finding that only the bacteria Bacillus subtilis was able to break down gliadin among the examined bacteria. It was also measured how much gliadin the bacteria could break down at different concentrations, and according to the data, it was evaluated that it is appropriate to adapt the culture to the presence of gliadin for a greater amount of broken gliadin.
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Biofilm formation in probiotic cultures and its application in pharmacy
Ryšávka, Petr ; Obruča, Stanislav (referee) ; Vorlová, Lenka (referee) ; Márová, Ivana (advisor)
The work was comprehensively focused on the development of adhesive forms of probiotics in the form of a biofilm on combined carriers with a prebiotic component. The second part dealed with the influence of food on the multiplication and survival of selected types of probiotic bacteria. Subsequently, the effect of individualized probiotic supplements on changes in the human intestinal microbiome was monitored. Suitable adherent probiotic strains for biofilm formation were selected and tested. Methods have been introduced and different variants of carriers for culturing and binding bacteria have been tested. In vitro experiments verified the stability of biofilm stucture and its resistance to low pH, bile and antibiotics in comparison with the planktonic cell form. The antimicrobial effect of probiotic strains in the form of a biofilm was studied. The cultivation of the multispecies biofilm on the combined carrier was optimized and the stability of the biofilm and the final viability of probiotic bacteria were confirmed. Furthermore, the influence of various foods and beverages on the viability of probiotic bacteria was evaluated with emphasis on the simulation of passage through the gastrointestinal tract. Both models, solutions with standardised concentrations of alcohol, sugar, salts, proteins or different pH and different types of real foods and beverages were tested. The effect of food and beverages was tested on monocultures of Lactobacillus acidophilus, Bifidobacterium breve and on probiotic capsules containing a mixed culture of probiotic microorganisms. The survival of probiotics in various food matrices in the simulated gastrointestinal tract was quantitatively different. We managed to define foods suitable for supporting the multiplication of probiotic bacteria. A separate part of the work was focused on the targeted modulation of the intestinal microbiome by individualized probiotics that were prepared on the basis of molecular biological analyzes of the intestinal microbiome aimed at detecting the percentage of lactobacilli, bifidobacteria and phylum Firmicutes and Bacteroidetes. Personalized probiotic supplementation confirmed the positive effect of this approach on microbiome changes, especially on the increase of the content of lactobacilli, bifidobacteria and the overall diversity of the microbiome.
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