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Molecular identification of probiotic bacteria in milk products form commercial yoghurt cultures
Horňan, Samuel ; Fialová, Lenka (referee) ; Smetana, Jan (advisor)
Lactic acid bacteria are considered as an important group of bacteria with probiotic effects, which are being widely used in the food industry or pharmacology. Identification and characterization of important probiotic strains play an essential role in the validation of probiotic products for commercial purposes. Their identification using molecular-biology techniques (most commonly PCR method) is one of the standard tools in commercial operations and services. The aim of this bachelor thesis is a literature review of probiotics and probiotic strains as well as a summary of current knowledge about the use of molecular biology techniques for identification of these bacteria with probiotic properties in dairy products. The experimental part of this work verifies the presence of probiotic bacteria declared on selected commercial dairy products using the polymerase chain reaction (PCR) method.
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Authenticity of natural plant component in cosmetics products
Kubalová, Michaela ; Fialová, Lenka (referee) ; Němcová, Andrea (advisor)
The purpose of this thesis was to study the authenticity of selected natural ingredients in cosmetic products. These were specifically cosmetic products that contained citruses, mint or lavender. Commercially available isolation kits were used for DNA isolation. The presence of plant origin DNA was verified by PCR method using primers specific for the ITS2 region of plants. The presence of limonene, a significant allergen contained in said plants, was determined in the samples by PCR method using primers for limonene synthase. At the same time, its presence was verified by HPLC method. In addition, two primers were tested for lavender and monitored for their efficacy, with no significant difference in the usage.
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Detection of probiotic bacteria in milk diary food products using PCR technique
Krempaská, Vladimíra ; Fialová, Lenka (referee) ; Smetana, Jan (advisor)
Probiotic bacteria play an important role in a healthy lifestyle. They help the consumer maintain the balance of intestinal microflora and prevent the overgrowth of harmful bacteria. Accurate identification and characterization of these probiotic strains is essential for research and the food industry. For exact identification, the use of molecular biological methods is necessary, thanks to which it is possible to validate probiotic products for commercial use. In this bachelor thesis, the DNA of probiotic bacteria was isolated from available dairy products. Two methods of isolation were used to isolate bacterial DNA, both of them provided sufficiently concentrated and high-quality DNA for further analysis by polymerase chain reaction (PCR). The presence of the Bacteria domain, genera Lactobacillus and Bifidobacterium were proved. Finally, the presence of Lactobacillus acidophilus species was also detected in the products.
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Influence of matrix type on the authentication of foodstuffs containing fruits
Kopková, Pavlína ; Strečanská, Paulína (referee) ; Fialová, Lenka (advisor)
Certain types of food, mainly the more expensive ones, are often adulterated to reduce their manufacturing price. However, this reduces their quality and can also have a negative impact on the health of the consumer. Children's fruit products are also targeted by fraudulent producers, where the declared fruit is most often replaced by a cheaper version. This work focuses on the detection of adulterated foods using various analytical methods, in particular PCR. The theoretical part focuses on the issue of food adulteration, the analytical methods used for detecting adulteration, and also on mango and banana which are determined in this work. The aim of this thesis was to determine what effect the type of matrix has on the determination of fruit components in food by PCR. Three types of matrix were used for this purpose - fruit puree, smoothie, and bars. An important task was to optimize the DNA isolation to achieve adequate purity and concentration of DNA. Then, the amplifiability of the obtained DNA was verified. The DNA isolates were then analyzed by multiplex PCR with primers specific for mango and banana. The results were verified by agarose gel electrophoresis. Subsequently, it was possible to determine that the fruit component in bars and fresh smoothies was the most easily analyzed by PCR and, on the contrary, the determination was problematic for puree. The instrumental part was focused on the determination of phenolic compounds in the products by HPLC. For this purpose, optimization of the extraction of phenolic compounds was necessary. This method was able to detect the presence of mangoes in all samples.
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Development and optimization of methods for detection of selected fruit species in foodstuffs
Šlosárová, Katarína ; Wikarská, Monika (referee) ; Fialová, Lenka (advisor)
Various types of adulteration of food products are becoming more frequent in the food industry than ever. That is no different for fruit products. This bachelor thesis deals with the detection of mango in fruit purées for infants through the use of qPCR and HPLC in three selected commercial products and their prepared replicas. Within the framework of molecular biology, different primers specific for mango were tested. Several primers, which were the most suitable for the demonstration of mango in fruit purées, were selected based on the course of the qPCR reaction, the size of the amplified product, specificity, and sensitivity. Using these primers, the presence of mango was proven in all six analysed samples. Regarding the instrumental part of the work, a rapid and simple method of extraction of phenolic compounds for their analysis by HPLC was tested, which has been previously successfully used on another plant matrix. This method was proved to be unsuitable for the extraction of phenolic compounds from fruit purées for the purpose of their authenticity verification using HPLC [1].
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Validation of probotic bacteria presence in fermeted plant food products
Péčiová, Bianka ; Fialová, Lenka (referee) ; Smetana, Jan (advisor)
Probiotic bacteria are an essential part of human nourishment which provide many health benefits. We consume them not only in traditional dairy fermented foodstuff, but also in the ever-expanding fermented plant products. The theoretical part summarized general knowledge about probiotic microorganisms, individual species used in technology of processing plant and dairy fermented products, and their influence on human health. The following chapters are devoted to various methods of identification of probiotic bacteria with emphasis on molecular diagnostics, specifically by PCR method. The experimental part is focused on the identification of lactic acid bacteria in a plant fermented product – in a fermented cabbage using 2 methods of isolation bacterial DNA and subsequent analysis by PCR method.
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Determination of selected red fruit species in plant-based food using multiplex PCR and instrumental methods
Vybíralová, Natálie ; Langová, Denisa (referee) ; Fialová, Lenka (advisor)
In many cases food is becoming the subject of adulteration, including fruit products that contain raspberries, strawberries and blueberries. This bachelor thesis is focused on the detection of strawberries, rapsberries and blueberries in model and commercial baby food products using multiplex PCR and HLPC. The theoretical part of this thesis is focused on composition of red fruits, their importance in human nutrition and especially about isolation of DNA from plant material. The aim of the experimental part of work was the analysis of selected commercial and model mixtures fruit purees containg raspeberries, strawberries and blueberries using instrumental and molecular biological methods. The results of these metods are compared. Commercial purees were bought in retail grocery shop. Model mixtures of these purees were prepared in our laboratory. DNA was isolated from fruit purees after and it’s amplifiability was comfirmed, it was successfully used in multiplex PCR to confirm the presence of raspberries, strawberries and blueberries in fruit purees. In the instrumental part, certain phenolic substences which are specific to red fruits were detected by HPLC in model and commercial mixtures.
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Detection of selected fruit species in plant-based foodstuffs through instrumental analysis and methods of molecular biology
Ondruch, Petr ; Langová, Denisa (referee) ; Fialová, Lenka (advisor)
Food is often subject to adulteration, including fruit products containing strawberries. This work deals with the detection of strawberries in model and real fruit products, so that possible falsification can be detected. The aim of the work was the preparation of model and real fruit products, their analysis using selected instrumental and molecular-biological methods and mutual comparison of the results of these methods. Model mixtures of fruit products were prepared and commercial ones were purchased. The content of polyphenols and flavonoids was determined by UV-VIS spectrophotometry, which were characterized in more detail by HPLC. DNA was isolated from the fruit purees, suitable primers confirmed the amplifiability and formation of a specific product for strawberry, and this product was verified by electrophoretic control. In the instrumental part, certain substances were detected by extraction from model and commercial mixtures and the HPLC method, and the amount of selected substances was determined. The lowest amount of strawberry DNA that could be detected by selected primers in the PCR method was 0.7 ng /µl, in commercial mixtures this DNA could not be detected. In the HPLC method, the strawberry was detected in all samples by focusing on a particular fingerprint and the amount of pelargonidin-3-glucoside in the samples. The instrumental method can be considered more suitable in this case.
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Revision of the electronic diabetic file at Clinic of Diabetology IKEM
Fialová, Lenka ; Blaha, Milan (referee) ; Provazník, Ivo (advisor)
Electronic diabetic record (EDR) is an electronic medical record used in Diabetology Clinics, Instittute for Clinical and Experimental Medicine (IKEM), Prague. EDR is complex record of patient statistical and clinical data. It´s complexity and large time demands on filling in all items make EDR not user friendly. Thus, benefits of electronic medical record are not fully utilized. The paper describes solution that leads to simpler user interface as component of outpatient message allowing easy entering data. The new interface has been designed and created in the form of program module. There are described other procedures to implement module into IKEM information system Zlatokop. At the end, there are discussed the advantages and shortcomings of this revised vision and current status of project of EDR. The first part of the work is about the basic concepts and problems regarding EDR. It is about e-health, specially about the electronic health record, databases, specially about Caché, which is used in Zlatokop, about diabetes mellitus and about user experience.
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Non-traditional sources of proteins and their potential applications
Kubisová, Petra ; Fialová, Lenka (referee) ; Skoumalová, Petra (advisor)
This bachelor thesis is focused on choosing the non-traditional sources of proteins, their characterization and subsequently its potential applications in diet of sportsmen. The theoretical part is focused on specification of importance of proteins in nutrition, description of different available protein supplements for sportsmen and essential characterization of chosen samples. The practical part of this bachelor thesis is focused on analysis of proteins, sacharides and lipids in samples. Nutritional components were analysed by different methods such as spectrophotometric methods, electrophoresis, gas chromatography etc.
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