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Microbial characterization of yoghurts and probiotic foods.
Kocourková, Hana ; Burdychová, Radka (referee) ; Španová, Alena (advisor)
Zvýšená konzumace fermentovaných mléčných výrobků v posledních letech vede k zavádění nových výrobků na trh. Funkční a probiotické produkty jsou v dnešní době také hojně rozšířeny. Cílem diplomové práce bylo posoudit mikrobiální kvalitu komerčních jogurtů a nefarmakologických probiotických výrobků. Bylo zkoumáno 24 vzorků jogurtů a 8 vzorků probiotických mléčných výrobků. Pozornornost byla zaměřena zvláště na kvantifikaci mikroorganismů v jednotlivých výrobcích a na sledování viability buněk. Většina mléčných produktů obsahovala počty odpovídající minimální požadované hodnotě 1x107cfu/ml. Nicméně byly objeveny produkty, které neobsahovaly živé bakterie nebo obsahovaly jen jejich menší množství. To platí zejména pro běžné jogurty. Probiotické mléčné výrobky v zásadě obsahovaly větší množství bakterií než jogurty, protože je u nich navíc požadováno 1x106cfu/ml minimálního množství specifických bakterií jiných než jogurtových startovacích kultur. Isoláty z fermentovaných mléčných výrobků byly identifikovány jako Streptococcus thermophilus and Lactobacillus spp. Isoláty byly poté rozlišeny na jednotlivé kmeny pomocí RAPD použitím tří různých primerů. Byla zjištěna velká různorodost mezi kmeny rodu Lactobacillus spp. používaných různými podniky.
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Molecular characterization of selected clostridial strains isolated from cheeses
Chroboková, Maria ; Kvasničková, Eva (referee) ; Rittich, Bohuslav (advisor)
The study was focused on molecular characterization of 42 clostridial strains. DNA was isolated by fenol-chloroform extraction procedure and precipitated with ethanol. After DNA isolation, PCR amplifications with specific primer sets were used for genus and species identification. Finally 19 strains were clasified as Clostridium tyrobutyricum and 3 strains were clasified as Clostridium butyricum. Presence of hydrogenase gene hydA was tested by PCR amplification using specific primer set HGf and HGr. Presence of hydrogenase gene was detected within 21 strains. (GTG)5 primer (rep-PCR) and Pr1 and Pr6 primers (RAPD) were used for differentiation of clostridial strains. Next, the cultivation of Clostridium tyrobutyricum S5 was studied under different conditions. The cultivation was carried out in liquid Reinforced Clostridial Medium (RCM) with lactose and cheese whey instead of glucose under anaerobe conditions. Growth was observed at laboratory temperature (20 to 23 °C) and at 37 °C, pH values ranging from 4.0 to 8.0 with 0.5 unit.
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DNA markery rezistencie voči biotickému stresu u uhorky siatej
Zvalová, Monika
Thanks to ever-advancing biotechnology techniques, DNA markers have become an integral part of the study of the plant genome. Knowledge of ideal molecular markers is widely applicable and helpful in genome mapping, genetic map construction, systematic taxonomy or conventional breeding. Compared to conventional approaches, DNA markers have many advantages, such as high stability, reliability and they are not affected by the environment. The diploma thesis deals with the application of various types of genetic markers in the study of regions of the cucumber genome responsible for resistance or susceptibility to selected biotic stressors, especially bacteries and viruses. It compares the effectiveness of RAPD with SCAR and CAPS markers. Furthermore, suitable and unsatisfactory genetic markers against angular leaf spot and disease caused by Zucchini yellow mosaic virus have been defined, which contribute to the economic losses of many crops every year. The work includes optimization of the PCR protocol and sequence analysis of selected amplified regions of the cucumber genome and their subsequent comparison in the NCBI database. For the future, a practical use of the results is proposed for cooperation with the MORAVOSEED CZ a.s company. The conclusion of the laboratory work can be helpful in the perspective use of markers specific to PSyLA and ZYMV.
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Rezistence genes against fungi deseases in cereals and methods for their detection
Stuchlíková, Šárka ; Dumalasová, Veronika (advisor) ; Schwarzerová, Kateřina (referee)
The paper deals with the possibilities of detection fungal disease resistance genes in the original European cereals, such as wheat, barley, rye and oats. It provides an overview of the most important resistance genes for breeding. The significance of the resistance genes is evaluated on the basis of the harmfulness of the individual cereal fungal diseases, as well as the importance of the individual cereal species. It describes the possibilities of detecting resistance genes using molecular markers and compares various types of molecular markers. It seeks answers to the question of where the breeding on resistance and therefore the use of molecular markers brings the greatest benefit.
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Molecular characterization of selected clostridial strains isolated from cheeses
Chroboková, Maria ; Kvasničková, Eva (referee) ; Rittich, Bohuslav (advisor)
The study was focused on molecular characterization of 42 clostridial strains. DNA was isolated by fenol-chloroform extraction procedure and precipitated with ethanol. After DNA isolation, PCR amplifications with specific primer sets were used for genus and species identification. Finally 19 strains were clasified as Clostridium tyrobutyricum and 3 strains were clasified as Clostridium butyricum. Presence of hydrogenase gene hydA was tested by PCR amplification using specific primer set HGf and HGr. Presence of hydrogenase gene was detected within 21 strains. (GTG)5 primer (rep-PCR) and Pr1 and Pr6 primers (RAPD) were used for differentiation of clostridial strains. Next, the cultivation of Clostridium tyrobutyricum S5 was studied under different conditions. The cultivation was carried out in liquid Reinforced Clostridial Medium (RCM) with lactose and cheese whey instead of glucose under anaerobe conditions. Growth was observed at laboratory temperature (20 to 23 °C) and at 37 °C, pH values ranging from 4.0 to 8.0 with 0.5 unit.
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Microbial characterization of yoghurts and probiotic foods.
Kocourková, Hana ; Burdychová, Radka (referee) ; Španová, Alena (advisor)
Zvýšená konzumace fermentovaných mléčných výrobků v posledních letech vede k zavádění nových výrobků na trh. Funkční a probiotické produkty jsou v dnešní době také hojně rozšířeny. Cílem diplomové práce bylo posoudit mikrobiální kvalitu komerčních jogurtů a nefarmakologických probiotických výrobků. Bylo zkoumáno 24 vzorků jogurtů a 8 vzorků probiotických mléčných výrobků. Pozornornost byla zaměřena zvláště na kvantifikaci mikroorganismů v jednotlivých výrobcích a na sledování viability buněk. Většina mléčných produktů obsahovala počty odpovídající minimální požadované hodnotě 1x107cfu/ml. Nicméně byly objeveny produkty, které neobsahovaly živé bakterie nebo obsahovaly jen jejich menší množství. To platí zejména pro běžné jogurty. Probiotické mléčné výrobky v zásadě obsahovaly větší množství bakterií než jogurty, protože je u nich navíc požadováno 1x106cfu/ml minimálního množství specifických bakterií jiných než jogurtových startovacích kultur. Isoláty z fermentovaných mléčných výrobků byly identifikovány jako Streptococcus thermophilus and Lactobacillus spp. Isoláty byly poté rozlišeny na jednotlivé kmeny pomocí RAPD použitím tří různých primerů. Byla zjištěna velká různorodost mezi kmeny rodu Lactobacillus spp. používaných různými podniky.
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Genetic variation and reproduction strategy of Gentiana pannonica in different habitats
EKRTOVÁ, Ester
The reproduction strategy and genetic variability of Gentiana pannonica, an endangered species in the Czech Republic, were studied. The differences of vegetation composition, seedlings recruitment, seed set and genetic variation within and among six various populations G. pannonica in the Šumava Mts. and two populations in the Alps of this species were investigated in a field. The vegetation composition differs between primary and secondary habitat of G. pannonica and seedlings recruitment was recorded only in primary habitat. The reproduction output of the study species is probably influenced by sufficient amount of pollinators. No serious genetic diversity between populations in the Šumava Mts. and the Alps was found. The genetic differences were found only between populations on the primary habitats and populations on the secondary mountain meadows.
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Genetic variability of natural populations of the \kur{Diuraphis noxia} (Aphididae)
SÁBOVÁ, Michala
Genetic variability of natural populations of the species Diuraphis noxia (Aphididae) was studied with RAPD method. First goal was to establish the chosen method in the laboratory and to optimize conditions of RAPD-PCR for D. noxia. This was followed by the pilot study, in which 12 populations were analyzed. Expected variability among individual populations and within population was acknowledged. The results of this investigation came up to the noted progress of diffusion of this species around the World. Results will be used in further more complex study of D. noxia.
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