Este trabalho teve como objetivo, determinar o rendimento dos extratos de folhas e cascas da espécie florestal Guazuma ulmifolia, identificar os principais componentes químicos dos extratos obtidos, e analisar a ação fungitóxica ao fitopatógeno Fusarium sp. Para a determinação da umidade, inseriu-se as amostras em formas de alumínio, posteriormente colocadas na estufa a 50ºC por 4 dias. Para avaliar o rendimento das amostras na extração com Soxhlet, utilizou-se os cones de filtros secos em estufa a 50ºC, e na maceração à frio avaliou-se o peso dos recipientes. Na representatividade das extrações, o álcool etílico apresentou maior rendimento para amostras de folhas e cascas. No bioensaio com o fitopatógeno, foram selecionados os extratos de folhas e cascas em hexano e acetato de etila, nas concentrações de 50.000μL/mL à 1.000μL/mL. As análises fitoquímicas foram realizadas em CG(M-S), utilizando o gás hélio (He), e os principais componentes detectados nos extratos em acetato de etila foram: Fitol, Esqualeno, Tetracontano, Lupeol e Tocoferol (Vitamina E) para as folhas, e Ácido Linolênico, Tocoferol, Esqualeno, Estigmasterol e o Fucosterol, para as cascas. / The objective of this work was to determine the yield of leaves and bark extracts of the Guazuma ulmifolia forest species, to identify the main chemical components of the extracts obtained, and to analyze the fungitoxic action of the phytopathogen Fusarium sp. For the determination of the humidity, the samples were inserted in aluminum forms, later placed in the oven at 50ºC for 4 days. In order to evaluate the yield of the samples in the Soxhlet extraction, the greenhouse filter cones were used at 50ºC, and in the cold maceration, the weight of the containers was evaluated. In the representativity of the extractions, ethyl alcohol presented higher yields for leaf and bark samples. In the phytopathogen bioassay, extracts of leaves and bark were selected in hexane and ethyl acetate at concentrations of 50,000 μL / mL at 1,000 μL / mL. The phytochemical analyzes were performed in CG (MS), using helium gas (He), and the main components detected in ethyl acetate extracts were: Fitol, Esqualeno, Tetracontano, Lupeol and Tocoferol (Vitamin E) for the leaves, and Linolenic Acid, Tocopherol, Squalene, Estigmasterol and Fucosterol, for bark

The effects of varying levels of sulphur dioxide (SO₂) on the cold maceration process was investigated with Pinot noir (Vitis vinifera L.) wine. The effects of these varying levels on the wines composition and colour parameters were examined. Cold maceration is a technique whereby grapes are crushed and placed at low temperatures (4 - lO°C) in the presence 50 - 150 mgL⁻¹ SO₂. This process is believed to provide a medium for the extraction of water soluble phenolic compounds, rather than the alcoholic extraction employed in normal fermentations. The extraction of these phenolic compounds was monitored from the juice through to six months of bottle age. The changes were measured using both Spectrophotometric and High Performance Liquid Chromatographic (HPLC) procedures. Cold maceration wines were found to be not significantly different to the control wine in all compositional parameters other than titrateable acidity which was found to be less than the control for all the cold maceration wines. The unsulphured cold maceration wine was not significantly different from the control wine in any of the spectral measurements except natural degree of ionisation, in which it was higher, and total phenolics, in which it was lower. These results indicate that the cold maceration process alone does not alter the extraction of phenolic compounds. The HPLC analysis of the wine confirmed the spectral results indicating that their were no significant differences in the levels of extraction of anthocyanins. The sulphured cold maceration wines were significantly greater than the control in visible colour, colour density, total anthocyanins, natural degree of ionisation, ionised anthocyanins and total phenolics. These results followed similar patterns with wine ageing, at six months these wines were still significantly greater in all the measurements apart from natural degree of ionisation. The results for the sulphured cold maceration wines indicates that SO₂ is acting as a solvent for the extraction of phenolic compounds including anthocyanins. The 50 mgL⁻¹ SO₂ cold maceration wine had similar colour and phenolic content to the 100 mgL⁻¹ SO₂ cold maceration wine at bottling, at six months the 50 mgL⁻¹ SO₂ cold maceration wine still retained a similar colour to the 100 mgL⁻¹ SO₂ cold maceration wine but had vastly reduced anthocyanin content. This indicates that for the grapes utilised in this study the most appropriate level of addition at cold maceration would be 50 mgL⁻¹ of SO₂. With grapes of differing phenolic content the level of addition required will vary.

Two pre-fermentation treatments were investigated in Pinot noir (Vitis vinifera L.) wines. The effects of cold maceration and carbonic maceration on the wines' composition, colour parameters and sensory properties were examined. Cold maceration is a winemaking technique used to increase non-alcoholic extraction in Pinot noir winemaking prior to fermentation. It involves holding crushed grapes with approximately 100-150 mg l⁻¹ SO₂ at low temperatures and is thought to increase the colour, aroma and flavour of the resulting wines. Carbonic maceration uses whole bunches that have undergone anaerobic metabolism to produce characteristically fruity and spicy wines. Pre-fermentation cold maceration produces wines that are higher in titratable acidity and monomeric anthocyanin content, but lower in colour density, hue and polymeric pigments. Reducing the maceration temperature below 10°C has little effect. Carbonic maceration produces wines that are lower in titratable acidity, monomeric anthocyanin content, and colour density but are higher in colour hue and amount of polymeric pigments. Quantitative descriptive analysis was used to define the effects of these pre-fermentation maceration treatments on the sensory characteristics of the resulting wine. Trained panel members found that there were no discernable sensory differences in the compositional parameters despite measurable chemical differences. Investigation into the aroma and flavour characteristics of the wines found that carbonic maceration produces wines that were lower in berry aroma and higher in acetate or ester-type aromas than the control wines. These wines were considered to have specific raspberry, floral, sugar, cherry and chemical aromas. This chemical note was also observed in the flavour of the carbonic maceration wines. The temperature of the cold maceration process has no major effect on the aroma and flavour of the resulting wines. However, the 10°C maceration was higher in woody/tobacco aroma than the 4°C maceration, and the 10°C treatment was also higher in bitter flavour than all the other treatments. Cold maceration wines were found to have specific mixed berry, dried fruit and sweet-oxidised aroma characters, together with a blackberry flavour note.

This research investigated yeast populations and diversity during pre-fermentation cold maceration and alcoholic fermentation of Vitis vinifera L. cv. Pinot noir grapes from a commercial vineyard (Dayton, OR). Fermentations were conducted at the Oregon State University research winery in 100 L tanks while grapes from the same vineyard lot were fermented at a commercial winery. Samples were taken daily during pre-fermentation maceration (9°C) and alcoholic fermentation (27°C) and plated on WL and lysine media to determine Saccharomyces and non-Saccharomyces populations and diversity. Total non-Saccharomyces populations increased from 1 x 10³ cfu/mL to 1 x 10⁵ cfu/mL during pre-fermentation cold maceration and reached a maximum of 1 x 10⁷ cfu/mL during alcoholic fermentation. Thirteen distinct yeast species were tentatively identified based on appearance on WL media and were initially screened for β-glucosidase activity using 4-methyllumbelliferyl-β-D-gluconopyranoside (4-MUG) plates. The identity of the isolates screening positive for β-glucosidase activity was determined by sequence analysis of the D1/D2 domain of the 26S rDNA gene. The five isolates identified were Metschnikowia pulcherrima, Hanseniaspora uvarum, Kluveromyces thermotolerans, and two Saccharomyces cerevisiae isolates. β-glucosidase activity was further characterized and quantified using a liquid media representing grape must conditions (pH 3.5, 20° Brix) at two temperatures (25°C and 8°C). While increasing sugar concentration suppressed the β-glucosidase activity of H. uvarum (-99%), β-glucosidase activity still remained relatively high for M. pulcherrima, S. cerevisiae isolate 1, and S. cerevisiae isolate 2. At 8°C, β-glucosidase activity was reduced for M. pulcherrima compared to activity at 25°C, but activity increased for K. thermotolerans, S. cerevisiae isolate 1, and S. cerevisiae isolate 2. The yeast isolates possessing β-glucosidase activity were used in fermentations of Vitis vinifera L. cv. Pinot Noir grapes. The grapes were treated with high hydrostatic pressure (HHP) to inactivate naturally occurring yeast and bacteria. All yeast isolates grew during pre-fermentation cold maceration (7 days at 9°C) and populations increased 3 to 4 logs. Following pre-fermentation cold maceration, all ferments were warmed to 27°C and inoculated with S. cerevisiae RC212. Alcoholic fermentations were all complete within eight days and after pressing wines were analyzed for volatile aroma compounds by SPME-GC-MS. The presence of different yeast isolates during pre-fermentation cold maceration resulted in wines with unique aroma profiles. Ethyl ester concentrations were highest in the wine that did not undergo a pre-fermentation cold maceration, while concentrations of branch-chained esters were higher in the treatments with yeast present during pre-fermentation cold maceration. Pre-fermentation cold maceration with yeast isolates demonstrating β-glucosidase did not affect the concentration of β-damascenone or β-ionone. Wines that had undergone pre-fermentation cold maceration with S. cerevisiae isolate 1, S. cerevisiae isolate 2, and a combination of all isolates resulted in over twice the concentration of β-citronellol over wines that did not undergo a pre-fermentation cold maceration. / Graduation date: 2013