This diploma thesis is focused on determination the aroma profile of selected species of fruit juices (apple, grape, carrot, orange, blackcurrant and elderberry juice). The theoretical part deals with the composition and properties of selected species of fruits and their juices, then deals with juice‘s technological process and selected methods for the determination of aroma active compounds. The experimental part describes the instrumental and sensory analysis. The six pure fruit juices and five mixed fruit juices were used for instrumental evaluation. The aroma active compounds were extracted by SPME and subsequently identified and quantified by gas chromatography. The sensory evaluation was performed by ranking test and evaluation using hedonic scale.

This master thesis deals with the assessment of aroma active compounds in fruits of currant (Ribes spp.). The theoretical part was focused on basic characteristics and chemical composition of currants, as well as the basic principles of techniques used: solid phase micro extraction (SPME), gas chromatography (GC) and sensory analysis. The main aim of the experimental part was to determine aroma active compounds in currants using SPME-GC-FID method. The profile and scale tests were used for assessment of sensory quality and tastiness of fruits. In total 18 varieties, among them 6 white, 6 red and 6 black currants, were evaluated. Fifty aroma active compounds were identified including 21 alcohols, 10 aldehydes, 2 acids, 8 ketones and 9 esters. The results were statistically evaluated using principal component analysis (PCA) to investigate the influence of single compounds on sensory quality of fruits.

The thesis deals with monitoring of the yeast's impact to the chemical and sensorial characteristics of produced wine. The aim of experimental part was obtaining of the aroma-active substances in the fermenting must and wine Hibernal and wine Pinot Blanc. SPME-GC-MS methods were used for identify wines and must aroma. The same autochthonous yeasts were used for wine production. This yeast was isolated from surface of grapes in our laboratory. It was found that both wines have a similar aromatic profile. The next aim was the monitoring of chemical parameters of samples and their changes. For this measurement were used spectrophotometric methods and HPLC methods. The last goal of the thesis was the isolation and identification of yeasts from wine Pinot Blanc by PCR-RFLP. These yeasts were then compared with yeasts isolated from Hibernal must. Identification of yeasts were done by amplification 5,8S-ITS DNA stretches using primers ITS1 and ITS4. Restriction endonuclease HaeIII, HinfI, HhaI and TaqI. were used for restriction analysis then.

Wood contaminated by polycyclic aromatic hydrocarbons (PAH) represent a serious environmental risk. Composting seems to be an effective method for decontamination of such waste material. PAHs are degraded mainly under aerobic conditions the efficiency of composting could be strongly affected by the process conditions. This work is aimed at the comparison of PAH removal by a 340-days-long composting in laboratory scale composting pilots and a 240-days-long process in a pilot scale. To set up composting piles a "green substrate" commercially used for button-mushroom production and a grass substrate were used. The use of the grass substrate led to a higher PAHs removal (97% of the initial sum of PAHs) in comparison to the green substrate (81% of the initial sum of PAHs). Further, the effect of the size of wood particles on the PAHs removal efficiency was demonstrated. Analysis of phospholipid fatty acids showed an enhanced growth of gramm-negative bacteria and fungi during the composting processes. During the degradation process the enzyme activity of laccase was detected. The ecotoxicological test using a bioluminescent bacteria Vibrio fischeri and a seed germination test using seeds of Hordeum vulgare L. showed that probably no toxic products were formed during the PAHs degradation in these experiments.

Aromatic pollutants pose a serious environmental problem. Petroleum and its derivates belong to the most abundant contaminants in the Czech Republic and their sanation is a priority objective for improving the life quality of the population. Bioremediation is a technology taking advantage of the natural capacity of soil and water microbial community to degrade environmental pollutants. Deeper understanding and detailed knowledge on specialized bacterial species, pathways and genes is required for selection, optimization and application of targeted bioremediation approach and for monitoring of its results. Recent analysis of a metagenomic library constructed from highly contaminated soil of the former military air-base Hradčany has identified a novel group of catabolic genes encoding extradiol dioxygenase similar to DbtC of Burkholderia sp. DBT1. The DbtC-like enzymes are among the three priority groups of extradiol dioxygenases with biodegradation relevance for the locality. The present study of soil bacterial isolates and metagenomic fosmid clones harboring the genes of interest gained evidence about the dbtC-like genes as a part of highly mobile gene cassette. Transposon insertion mutagenesis identified the genes joined with the expression of the extradiol dioxygenase activity. The dbtC-like genes were...

Dissertation thesis Study of physical chemical features in fruits of peaches and plums was developed at the Institute of Post Harvest Technology of Horticulture Products at Faculty of Horticulture of Mendel University in Brno. This thesis concentrated on setting of material elements in fruits of peaches, the variety 'Red Haven' in the period of ripening and post-harvest storage in different types of atmosphere according to contents of CO2 and O2. In five analysed varieties of peaches 'Flamingo', 'Harbele', 'Harblaze', 'Harbrite', and 'Maria Marta' were set ripening values during ripening on tree. In varieties of plum fruits 'Valjevka', 'President', 'Stanley' was found their tolerance against gas mixtures. During ripening of peach varieties were set volatile elements (ethanol, acetaldehyde) and was stated the concentration of ethylene in the critical period of ripening. In literary review there are characterized material elements of pulp fruits, both chemical structure of fruits, and volatile aromatic substances and their biogenesis, there are described in detail material substances of fruits, amongst which belong biological, physical, sensory, technological signs of quality and post harvest influences on fruits. Further ethylene is characterized in post-harvest period and its physiological features and methods of its effect limitation, factors of plant reaction against ethylene, and chemical inhibition of ethylene reception. Next part deals with methods of quality evaluation and techniques of fruit storage, further on with fruits of plums, peaches and their general characteristics. Experimental part evaluates several fractional methodics. Assessment of peach varieties in three stages of ripening and setting of ripening index, influence of low content of oxygen on production of anaerobic metabolites and volatile materials stated in plum fruits, distribution of frequency of peach fruits ripening on tree in five stages of ripening, production of ethylene according to the grades of ripening in peach varieties, their sensory evaluation and setting of volatile aromatic compounds. In variety 'Red Haven' there were evaluated grades of ripening in the range: unripe to over ripe, which represented in dates five successive harvests. Each successive term was compiled of 40 fruits of average weight for the set grade of ripening. Production of ethylene was evaluated through distribution of fruits in settled harvest date, and was distinctively different in fruit distribution according to ethylene content pursuant to harvest date. In executed measurements of fruits of peach variety 'Red Haven' ripening on tree in grades of ripening unripe, ripe, over ripe, in their frequency five to seven there was stated concentration of ethanol and acetaldehyde expressed in mg*l-1 by direct spraying of juice into filling analytic colon. Concentration of acetaldehyde did not change during fruit ripening, and this metabolite can be assessed as interim which changed by alcohol dehydrogenase into ethanol. Concentration of ethanol in over ripe fruits is characteristic with higher content which does not represent multiple concentrations in over ripening in common oxygenated atmosphere. Evaluation was executed only in one variety, however, similar tendencies should be supposed in other varieties as well. Index of ripening (IZ = RS* strength/TK) was formulated in five peach varieties, which represents processes of fruit ripening on tree and important differences were found for ripe and unripe fruits, whereas index of ripening between ripe and over-ripe fruits differs only in several units. With following ripening rates in varieties there were determined differences in phases unripe to ripe for varieties 'Harblaze' and 'Maria Marta'. Furthermore these varieties were evaluated by sensory analysis as well. In analytical method HP/GC/MS 42 volatile aromatic compounds were specified in groups of alcohols, aldehydes, esters, terpenes, lactones, and organic acids in plum fruits in varieties 'Valjevka' and 'Stanley'. In Tukey´s test productions of each specified volatile compounds were evaluated. In variety 'Red Haven' treated with four different gas mixtures (ULO, CA, RA, FAN) according to percentage of content of set amount of CO2 and O2, and stored in these atmospheres during 29 days, there were specified several volatile aromatic materials through analytical method HP/GC/MS.

In the present dissertation thesis quality of plum cultivars were investigated in three successive experiments included measurement of ethylene, CO2, ethanol, acetaldehyde, firmness of the fruits and determination of volatile aromatic compounds by SPME/GC/MS Two plum fruit varieties (Stanley and Valjevka) were stored in prepared gas mixtures (FAN: 0,6 % O2 a 1 % CO2, CA: 1 % O2 a 10 % CO2, RA: 21% O2 a 0,03 % CO2) for 70 days. In terms of the 20th, 40th and 60th day followed shelf life for 10 days at 20 °C. Firmness of the skin was different for both varieties and the softening increased in the variety Valjevka. Softening of fruits significantly decreased in FAN and CA. Skin firmness of fruits stored in normal oxygen atmosphere was lower for about 0,59 MPa (Stanley) and 0,67 MPa (Valjevka) at the end of storage. Fruits stored in CA atmosphere were firm, with unbrowning flesh and there was no browning in stem area, too. The results showed that the variety 'Valjevka' has a high capacity (1705 mg/l) to produce ethanol.There was no microbial damage in 'Stanley' stored in CA and FAN at the end of the storage, in RA were 13,19 %. The highest microbial attack was observed in Valjevka stored in RA 41,85 %. Seventy-one volatile compounds were sampled by head space SPME in two plum fruit varieties (Stanley and Valjevka). Using PCA analysis 13 compounds are sufficient for distinguishing ripening of fruits variety Stanley treated by tree storage atmospheres ((Z)-hex-3-en-1-ol, 2-methyl butan-1-ol, (Z)-3-octen-1-ol, 2-heptanol, heptanal, (Z)-3-hexenal, (E)-2-decenal, n-octanal, ethyl acetate, (E)-2-hexenyl acetate, a-terpineol, b-cyclocitral, b-ionone) and 11 compounds for 'Valjevka' (2-octanol, n-octanal, (E)-2-hexenyl acetate, b-cyclocitral, b-ionone, (Z)-3-octen-1-ol, 2-methyl-1-pentanol, (Z)-3-hexenal, n-hexanal, ethyl benzoate a butyl 2-methylbutyrate). The fruits of 13 plum cultivars were analysed at two different stages of maturity: first when they were ready for picking according to conventional commercial criteria, and again after seven days of maturation during shelf life. In the period of over-ripening SSC increased, and no differences were found in relation to the cultivar. The fruits in this period were physiologically in a phase of reduced intensity of respiration, while production of ethylene increased and was associated with the earliness of the cultivar. Cultivars with a short vegetation period produce more ethylene while late-maturing cultivars have a low potential for ethylene production. Based on post-harvest changes in the tested quality factors, the late-ripening plum cultivars (cv.) Jojo, Topend, President, Tophit and Elena have higher storage potential. A high significant negative correlation was detected between levels of skin firmness and ethylene production. 69 of volatiles were determined by GC/MS/SPME. There are eight compounds (oct-1-en-3-ol, p-menth-1-en-4-ol-R, n-hexan-1-ol, n-pentan-1-ol, ethylhexan-1-ol, g-nonanolaktone, g-kaprolaktone and g-oktalaktone) which taken together can be used to distinguished the two different stages of maturity. The fruits of cv. 'Tophit' were untreated or treated with 1-MCP for 24 h at 20 °C and then kept at 1 °C for 50 days in the atmosphere CA (2 % O2 a 7 % CO2) and RA. Plums were evaluated after 10, 30, 50 days of storage at 1 °C followed by shelf life storage for 7 days. The effect of cold and 1-MCP treatment resulted in significant inhibition of ethylene production, respiration rate and retention of flesh firmness after transfer to room temperature. Sixty-four volatile compounds were sampled by head space SPME and identified by GC-MS, from which (E)-2-hexenal, (Z)-hex-3-en-1-ol, butyl-2-methylbutyrate and limonen were significant.

The former military air-base Hradcany is among the most contaminated with organic pollutants localities in Czech Republic. Main cleanup strategy in the area is the bioremediation taking advantage on the natural potential of the autochthonous soil microorganisms to evolve catabolic pathways for in situ degradation of the pollutant. The diversity and abundance of the pathways, as well as the specificity and activity of the encoded enzymes are priority biotic factors determining the bioremediation efficiency. Main task of this work was to analyze the bacterial diversity in jet fuel contaminated soils based on key catabolic genes encoding the Rieske non-haem iron dioxygenases of the toluene/ biphenyl oxygenase branch. High molecular soil DNA was extracted and the sequences encoding catabolic genes were selectively enriched by hybridization to biotinylated oligonucleotides on magnetic microbeads with covalently bound streptavidin. Fragments of the genes for the -subunits of Rieske non-haem iron oxygenases were amplified and analyzed by restriction analysis, cloning and sequencing. Their evolutionary histories were inferred using the Neighbour-Joining and the maximum likelihood methods. The catabolic genes diversity in the actively bioremediated and highly polluted soil HRB was compared with the diversity in the...

This thesis deals with the production and characterization of cheese analogues, ie products in which some of the dairy components are mutually or completely replaced by a non-dairy component. Model samples of fresh cheese and analogues of fresh cheese (standard) with the addition of linseed and rapeseed oil were produced by a standard technological procedure in MENDELU Brno. The content of bound and free fatty acids, volatile (aroma) substances and sensory quality was monitored in the samples. The main focus was to evaluate the effect of added vegetable oil on the monitored parameters. The fats from the sample were extracted with a mixture of diethyl ether and petroleum ether. Fatty acids were converted to methyl esters by boron trifluoride-catalyzed acid esterification and subsequently identified by GC-FID. The analysis found a higher content of fatty acids in cheese analogues in comparison with the control sample, also the content of unsaturated fatty acids increased. The HS-SPME-GC-MS method was used for the identification and semi-quantification of volatile substances (aromatic substances). 56 volatile compounds were identified in the samples; The percentage of individual chemical groups differed significantly, the highest content was found in the control sample. Sensory evaluation was performed using a scale and a serial test. In all of monitored indicators (apparence, color, consistency, smell and taste) the control sample was evaluated as best. As for the analogues the taste and smell of used oils were detected, which was unpleasant for the evaluators. The obtained results show that the addition of vegetable oils increases the nutritional value of cheeses (higher content of unsaturated fatty acids), but reduces their sensory quality and overall consumer acceptability.

This paper deals with the manufacture and characterisation of cheese analogues, which are products, where the milk components are totally or partially replaced by non-milk components. Model samples, edam type cheese and the analogue containing hazelnut oil, were manufactured using the standard way at Brno Mendelu. The main parameters chosen to describe the quality of samples were free and bound fatty acids, volatile compounds and sensory quality. The main aim was to judge the influence of added hazelnut oil on mentioned parameters. For the identification and semiquantification of volatile compounds HS-SPME-GC-MS method was used. Fat extraction from sample was made with a mixture of two solvents (diethylether, petrolether), fatty acids were identified and quantified by GC-FID, with their former acid transesterification using methanolic solution of BF3 as a catalyst. For sensory analysis of the appearance, colour, texture, aroma, taste and overall acceptance of the sample descriptive scales and comparison with standard were used, the structure was observed by electron microscopy. The results show that addition of hazelnut oil affects the sensory properties, ripening process (higher amount of volatile compounds, faster lipolysis), nutrition value (higher amount of unsaturated fatty acids), and the stability of the product.