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Evaluation of Physical and Chemical Parameters of Still Less Exploited Species of Small Fruit and Proposal of a New Soft Drink from this Fruit
Cetkovská, Jitka ; Krška,, Boris (referee) ; Barták,, Petr (referee) ; Omelková, Jiřina (advisor)
This thesis deals with still less expolited species of small fruit, evaluation of its physical and chemical parameters and proposal of a new soft drink from one of the studied species of furit. Firstly, for the characterisation of basic nutrition parameters (dry matter, content of organic acids and saccharides) and biologicaly active substances (vitamin C, anthocyanins and phenolic compounds) suitable titration, spectrophotometric, chromatographic and electrophoretic methods were chosen, optimized and validated. The sample preparation procedure was optimized as well. For three years (2010–2012) the content of listed parameters was monitored in some cultivars of five species of small fruit: in seven cultivars of sea buckthorn, in ten cultivars of cornelian cherry, in twelve cultivars of rowanberry, in two cultivars of chokeberry and in seventeen cultivars of elderberry. By means of Duncan’s test, analysis of principal components (PCA) and cluster analysis (CLU) the main differences between studied cultivars were observed mainly in sea buckthorn and rowanberry, where single cultivars were easily distinguishable from others on the basis of genetic origin. On the basis of comparison of determined parameters in studied species of fruit and with collaboration with fruit processing company the suitable species of fruit for commercial utilization was chosen – elderberry, as a important source of anthocyanins and phenolic compounds. A new soft drink as a proposal of utilization of this fruit was created – fruit juice or nectar with portion of elderberry juice. Mixed juices and nectars with various composition were prepared from elderberry and grape juice and the sensory analysis of these drinks was performed. The best evaluated were 100% fruit juices prepared with 30–50% portion of elderberry juice. The composition was jurally protected. On the basis of this protection the product consisted from 100% fruit juice with 10% of elderberry component mixed with apple and grape juice was introduced to the market by the fruit processing company.
Properties and production of microbial lipases
Martinková, Patrícia ; Reichstädter, Marek (referee) ; Omelková, Jiřina (advisor)
Bachelor´s thesis is focused on testing the culture media for growing various strains of yeasts, producing lipolytic enzymes and to study the influence of culture media composition on the production of lipolytic enzymes. Theoretical part of this thesis states the characteristics of lipolytic enzymes, their properties, sources, conditions for their production and the possibilities of their application. Together 8 strains of yeasts were used in the experimental part of the thesis, namely 2 strains of Yarrowia lipolytica, 2 strains of Kluyveromyces lactis, Cryptococcus saitoi, Candida intermedia, Candida oleophila and Debaryomyces hansenii, which were cultivated on 4 media with different compounds. During the cultivation the growth of biomass was monitored and growth curves were formed based on these results. Selected strains of yeasts were tested during the process of cultivation for lipolytic activity of two types of enzymes – enzymes bonded to the cellular structure and extracellular enzymes, which were measured by spectrophotometer. Production of lipolytic enzymes varied depending on applied culture medium.
Identification of yeasts from interspecific varieties of grapes
Sadel, Peter ; Omelková, Jiřina (referee) ; Vránová, Dana (advisor)
The main goal of my diploma thesis was to identify and characterize yeasts from must Hibernal and also collection yeasts by using methods called RFLP-PCR (Restriction Fragment Length Polymorphism - Polymerase Chain Reaction). Theory was the first part of my diploma thesis which dealt with wine, yeasts and molecular methods. Theory section was followed by experimental section divided into two parts. The main goal of the first part was to characterize and identify yeasts from must Hibernal by using PCR and RFLP-PCR methods. In the samples there were found yeasts Saccharomyces and Pichia. The second experimental part of my diploma thesis had a goal to extend the database of new yeasts using the same methods mentioned in the first part of experimental section.
Fragrance Allergens in Foods and Everyday Use Products
Divišová, Radka ; Buňka, František (referee) ; Hojerova,, Jarmila (referee) ; Omelková, Jiřina (advisor)
Fragrances are added to food, cosmetics and other products primarily for enhancement of their sensory quality (taste and/or aroma). However, the usage of these compounds is associated with the wide range of various adverse biological effects. Some fragrances widely used in cosmetics are proved to cause e.g. skin sensitization, rashes, dermatitis, headache, cough etc. To protect the health of consumers, European union approved the cosmetic directive (ES/1223/2009) that requires the labeling of 26 allergen fragrances on the final product label, if concentration exceeds the given level (0,01 % for rinse-off and 0,001 % for leave-on product). However, some producers hide the presence of allergens in the final product under the general term „aroma“. For this reason it is very important to monitor the content of these substances in cosmetic products. Fragrances are found not only in cosmetics, but the various types of food are also aromatized by them. Unlike cosmetics, the food producers are not obliged to label the presence of fragrance allergens on the packaging. Therefore, the monitoring of these fragrances is highly desirable because of the potential health risks they pose. The aim of this study was to develop a method for simultaneous determination of regulated fragrance allergens in food and everyday use products. The method based on extraction of analytes by solid phase microextraction coupled to gas chromatography with FID detection (HS-SPME-GC-FID) was chosen on the basis of the literature review. The fiber CAR/PDMS provided the highest extraction efficiency among the SPME fibers tested. Univariate and multivariate data analysis were used to optimize the main parameters affecting microextraction process. The final method validation was performed in terms of linearity, repeatability, reproducibility, recovery, limits of detection and quantification. The optimized and validated method was applied to a wide range of products including cosmetics, aromatized food and fragranced toys. These products were also subjected to sensory evaluation especially in terms of taste and/or aroma (i.e. flavour), which may be associated with the content of the monitored fragrances. The profile test (EN ISO 13299) and seven-point category ordinal scale (ISO 4121) were used for sensory evaluation.
Monitoring of Phytoestrogens Expression to the Milk and Milk Products
Watzková, Jarmila ; Rittich, Bohuslav (referee) ; Doležal, Petr (referee) ; Omelková, Jiřina (advisor)
Phytoestrogens are very intensively studied in recent years because they show different pharmacological effects on human health. For example, it was demonstrated that phytoestrogens reduce the risk of certain cancers (breast, prostate or colon), act as antioxidants in the body and affect the activity of certain enzymes. In addition, also it was demonstrated number of positive effects of soybean phytoestrogens on the health status of women in the menopause, such as reducing the risk of atherosclerotic cardiovascular disease or osteoporosis. Various technologically processed soy beans are important part of the feed rations mainly for high-lactating dairy cows, because they are a valuable source of the energy and high-quality protein, despite their negative effect on reproductive performance. Phytoestrogens consumed from the feed rations are hydrolyzed in the rumen of dairy cow and together with their metabolites are absorbed in the small intestine into the blood. They can be excreted from the body in urine or milk. From this perspective, cow's milk may become a source of phytoestrogens substances, especially equol in the human nutrition. On the other hand, some research results suggest possible potential risks associated with phytoestrogens intake in the diet on human health. At the some age (infants and young children) and group (vegetarians) categories may be dietary intake of phytoestrogens undiserable. According to most experts, a beneficial effect on human health prevail the potential risks. However, the risks associated with phytoestrogens intake in food can not be ignored and therefore phytoestrogens are still actual problem in the field of scientific research.
Isolation, Identification and Characterisation of Microbial Communities of Wine and Selected Foods
Šuranská, Hana ; Španová, Alena (referee) ; Jarošová, Alžběta (referee) ; Omelková, Jiřina (advisor)
Proposed dissertation thesis deals with wine and artisanal cheeses microbiology. The first part is focused on identification of yeasts isolated from grapes and musts during production of white and red wines. The grape varieties were grown under the integrated and organic farming on Moravian vineyard. Yeasts were identified by ITS-PCR-RFLP method (amplifying internal transcribed spacer ITS: ITS1, ITS2 and 5.8S rDNA) and unknown species were subjected to partial sequencing of ITS rDNA region. In total, 524 isolates were divided into 14 different species belonging to six genus were identified from. The first stages of fermentation process were characterised by predominance of non-saccharomyces species especially H. uvarum. Due to increased ethanol concentration strains of S. cerevisiae prevailed in the later phases of the process. Further, partial aim of this study was to isolate and to apply selected autochthonous S. cerevisiae strains as starter culture during controlled industrial wine fermentation process. Genus Saccharomyces was distinguished from other non-saccharomyces species by ITS-PCR-RFLP. Further, in order to distinguish Saccharomyces genus at the species and the strain level, several molecular methods were applied including PCR-fingerprinting (rep- and RAPD-PCR), species-specific primers (multiplex and touchdown PCR), LSU-DGGE and interdelta PCR. Species-specific primers enabled us to distinguish some species of the Saccharomyces sensu stricto complex. Furthermore, interdelta PCR seems to be useful tool for S. cerevisiae strains identification. Among 120 isolated autochthonous strains belonging to Saccharomyces genus, 45 different strains were identified. Based on its sufficient technological properties (osmo- and ethanol tolerance, low H2S production etc.), S. cerevisiae 1-09 strain isolated from grape berries coming from moravian vineyard was chosen. Strain S. cerevisiae 1-09 was tested in small amount of must and after that also during industrial fermentation of red and white wine production. Based on the results of chemical and sensorial analysis, the strain seems to be suitable for application as the starter culture for winemaking process. The final part of this thesis is focused on quantification and identification of the yeasts isolated from artisanal cheeses and their by-products coming from Western Balkan Countries. Isolated species were identified by ITS-PCR-RFLP, partial sequencing and by physiological tests. Among the 20 yeast species found, D. hansenii, C. zeylanoides and Y. lipolytica were found to be predominant. Moreover, we developed culture-independent, semi-quantitative technique based on construction of ITS-clone library from metagenomic DNA to investigate complex fungal communities associated with artisanal cheeses and their by-products. Novel technique is based on direct extraction of total DNA from the sample. This was compared with culture-dependent ITS-PCR-RFLP and culture-independent LSU-DGGE methods. The results highlighted the discrepancies among these methods. Finally, the divergences among applied methods were confirmed by correlation analysis and by indices of general biodiversity and dominance of species. ITS-clone library approach combines the advantages of cultivation-based analysis and LSU-DGGE with semi-quantification of fungal species without the requirement of their cultivation. This study might open new perspectives in direct and complex analysis of yeasts and moulds in food matrices.
Prospects of Microbial Degradation and Waste Utilization from Food Processing Industry
Illková, Kateřina ; Karovičová, Jolana (referee) ; Jarošová, Alžběta (referee) ; Omelková, Jiřina (advisor)
This work deals with the problem of microbial degradation of the waste materials from food industry. This work is focused on the production of technological significant enzymes producing by microorganisms, which were able to use the waste as a sole carbon source. In the first part of this work, the attention was focused on the production of pectolytic enzymes. This part was made within study interships in Slovak Academy of Sciences, department of Glycomics in Bratislava. The grape pomace as the waste form winegrowing was used as a sole carbon source for microbial growth and enzymes production. The production of pectolytic enzymes was tested on this waste. After screening the most suitable microorganisms was chosen with the highest production of polygalacturonase activity. Produced enzymes were isolated by extraction techniques, purified and then identified proteomically. The aim of the second part of this work was the waste water treatment containing lipids and lipolytic enzymes. The reason was the cooperation with the company constructing grease traps. The characterization of supplied commercial preparations was the subject of this work and the other reason was the characterization of conditions for lipases secreting by microorganisms, identification of microorganisms present in the commercial preparation and testing of new microbial cultures for the development of new preparation for the grease traps.
Multi-experimental Characterization of Grape Skins' Extracts
Šťavíková, Lenka ; Brezová, Vlasta (referee) ; Šilhar,, Stanislav (referee) ; Omelková, Jiřina (advisor)
The determination of anthocyanins in red grapes and wines has been of increasing interest in the last years, as they play an important role in colour quality of red wines revealing also many human health beneficial effects. They contribute e.g. to the reduction of coronary heart disease, but exhibit also antimutagenic, anti-inflammatory, anti-carcinogenic and antioxidant properties. In this doctoral thesis, the complex study of grape skin alcoholic and water extracts, prepared from Alibernet and St. Laurent wine grape varieties is presented. Extracts were prepared from three different amounts (0,5; 1,0 and 1,5 g) of lyophilized grape skin powder using the Pressurized fluid extraction (PFE) and the Pressurized Hot Water Extraction (PHWE) at different temperatures ranging from 40 up to 120°C and pressure of 15 MPa. Methanol, ethanol and water were used as a solvents. Total phenolic compound content (TPC) of individual extracts was determined using the Folin-Ciocalteau assay and their tristimulus color values (CIE Lab) were estimated, using the UV-VIS spectrophotometer. The identification and quantification of anthocyanins by high-performance liquid chromatography with diode array detection (HPLC-DAD) was performed. In addition, pH values of all extracts were also measured using the combinated glass electrode. Antioxidant activity of extracts was tested by EPR spectroscopy in Fenton system (H2O2/Fe2+) generating reactive radicals (•OH, O2-•, •CH3) followed by spin trapping technique, using 5,5-dimethylpyrroline-N-oxide (DMPO) as spin trap. In addition, radical scavenging activity of extracts was assessed applying 2,2-diphenyl-1-picrylhydrazyl (•DPPH) free radical and 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) cation radical (ABTS•+) assays. All the experimental data were processed with principal component analysis (PCA) and canonical discriminant analysis (CDA) to specify the optimum extraction conditions for extract preparation from the perspective of the potential further application of the extracts as food supplements or food colour enhancers. The results indicated that grape skins of both varieties are a promising source of anthocyanins with prospective application in food industry.
Determination of Natural Compounds in Select Plant Materials and Less Obvious Kinds of Fruit
Hohnová, Barbora ; Márová, Ivana (referee) ; Ventura, Karel (referee) ; Omelková, Jiřina (advisor)
Flavonoids are natural compounds widely distributed in plant kingdom. They are inseparable from human diet because they showed a protective effect against cancer, stroke and coronary heart diseases related to their antioxidant properties. Therefore, rapid and efficient extraction procedure prior to chromatographic analysis is required. The liquid extraction at elevated temperature and pressure – Pressurized fluid extraction (PFE) and Pressurized hot water extraction (PHWE), present fast, effective and environmentally friendly extraction methods for the determination of flavonoids in plant materials. PFE and PHWE followed by reversed phase high-performance liquid chromatography with UV-visible detection have been utilized for the determination of a group of flavonoids (rutin, myricetin, quercetin, luteolin, apigenin and kaempferol) in the leaves and berries of less common plants. The matrices were extracted by methanol, ethanol and water at higher temperature 40-120 oC and pressure 15 MPa during 15 minutes. The obtained results were compared with conventional Soxhlet extraction and ultrasound-assisted extraction, the same solvents were used. The most effective extraction of selected flavonoids was achieved by PHWE. PFE showed the extraction yields comparable to those of the Soxhlet extraction, and the lowest extraction power was displayed by ultrasound-assisted extraction.

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