A indústria do processamento de frutas gera um grande volume de resíduos. Entretanto, muitos destes resíduos podem apresentar elevado valor nutricional, ou ainda, apresentarem teores importantes de compostos ativos com atividades antioxidantes, podendo, em alguns casos serem superiores às concentrações encontradas na polpa da fruta. Entretanto, o descarte de tais resíduos pode provocar sérios problemas ambientais. Diversos estudos relatam que os resíduos agroindustriais provenientes do processamento (produção de sucos, polpas congeladas, extratos secos, etc) de frutas podem ser usados na produção de diversos produtos, tais como: produtos de panificação, ração animal, fertilizantes, produção de biogás, dentre outros. Por outro lado, ainda há poucos estudos e aplicações frente ao grande potencial apresentado por estes resíduos. Diversos estudos relatam que resíduos agroindustriais provenientes do processamento do camu-camu apresentam teores significativas de compostos ativos e elevado valor nutricional. Assim, o objetivo deste projeto foi produzir e caracterizar farinhas obtidas em diferentes temperaturas (50, 60 e 70°C) do resíduo do despolpamento do camu-camu (FRCC) e avaliar o potencial para aplicação das mesmas em cookies. O resíduo foi submetido a secagem em três diferentes temperaturas (50, 60 e 70°C) em estufa de circulação forçada de ar, trituradas em moinho de facas e a granulometria padronizada em 16 mesh. As farinhas obtidas, assim como o resíduo úmido do processamento do camu-camu foram caracterizados quanto: ao conteúdo de água, atividade de água, pH, cor, composição centesimal (proteína, fibras, lipídeos, cinzas e carboidratos), composição mineral (nitrogênio, fósforo, potássio, cálcio, magnésio, enxofre, boro, cobre, ferro, manganês e zinco), fenólicos totais, potencial antioxidante (métodos do DPPH•, FRAP e ORAC), carotenóides, taninos, vitamina C, capacidade de absorção de água e óleo, capacidade emulsificante e capacidade espumante. Após a caracterização, a farinha com potencial antioxidante superior e maior concentração de fenólicos totais, foi utilizada para a produção de cookies com as seguintes porcentagens de substituição da farinha de trigo: 5%(C-5FRCC), 10% (C-10FRCC), 15% (C-15FRCC) e 20% (C-20FRCC), além do controle (sem substituição).Por sua vez, os cookies foram avaliados em relação ao conteúdo de água, pH, cor, perda de água, altura, diâmetro, fator de espalhamento, volume específico, dureza, fenólicos totais e potencial antioxidante (métodos do DPPH•, FRAP e ORAC). Verificou-se, de um modo geral, que a temperatura de secagem afetou as propriedades das farinhas, sendo que as farinhas obtidas após secagem a 70°C (FRCC70) apresentou valores superiores de concentração de compostos ativos e potencial antioxidante, em relação às farinhas obtidas após secagem nas demais temperaturas. Desta forma, a FRCC70 utilizada na produção dos cookies. A substituição parcial da farinha de trigo pela FRCC70 provocou pequenas alterações nas características físicas (altura, diâmetro, fator de espalhamento, volume específico) dos cookies, principalmente para as porcentagens de substituição mais elevadas desta farinha. De um modo geral, os cookies com substituição pela FRCC apresentaram dureza superior. Verificou-se ainda que o aumento da concentração da FRCC70 nos cookies provocou aumento da concentração dos compostos fenólicos. Da mesma forma, verificou-se que o aumento da porcentagem de substituição da FRCC70 nos cookies aumentou o potencial antioxidante medido pelos métodos do FRAP e ORAC. Por outro lado, não verificou-se efeito do aumento da porcentagem de substituição no potencial antioxidante medido pelo método do DPPH• dos cookies. Assim, verificou-se que a farinha do resíduo do processamento do camu-camu mostrou potencial para aplicação em cookies por poder agregar valor nutricional, em função do aumento do potencial antioxidante e da concentração de fenólicos totais desses produtos. / Fruit processing industry generates a high volume of by-products. However, many of these byproducts may present high nutritional value, or they may present important concentrations of active compounds with antioxidant activities, and in some cases they could have higher concentrations than thosefound in the fruit pulp. However, the discard of such by-prodcuts can cause serious environmental problems. Several studies report that the agroindustrial by-products from fruit process (production of juices, frozen pulp, dried extracts, etc.) can be used in the production of several products, such as: bakery products, animal feed, fertilizers, biogas production, among others. On the other hand, there are still few studies and applications in view of the great potential presented by these by-products. Several studies reported that agroindustrial by-products from camu-camu processing have significant concentrations of active compounds and high nutritional value. Thus, the aim of this project was to produce and characterize flours obtained at different temperatures (50, 60 and 70 ° C) of the camu-camu pulp by-product (FRCC) and to evaluate the potential for application of the same in cookies. By-product was dried in three different temperatures (50, 60 and 70 ° C) in a forced air circulation oven, milled with a knife mill and the standard granulometry in 16 mesh. Flours and the wet by-product were evaluated in relation to water content, water activity, pH, color, centesimal composition (protein, fiber, lipids, ashes and carbohydrates), mineral composition (nitrogen, phosphate, potassium, calcium, magnesium, sulfur, boron, copper, iron, manganese and zinc), total phenolics, antioxidant potential (DPPH•, FRAP and ORAC methods), carotenoids, tannins, vitamin C, water absorption capacity and oil, emulsifying capacity and foaming capacity. After the characterization, the flour with higher antioxidant potential and higher total phenolic concentration was used to produce cookies with the following percentages of wheat flour substitution: 5% (C-5FRCC), 10% (C-10FRCC), 15% (C-15FRCC) and 20% (C-20FRCC), in addition to the control (without substitution). Cookies were evaluated in relation towater content, pH, color, water loss, height, diameter, spreading factor, specific volume, hardness, total phenolics and antioxidant potential (DPPH•, FRAP and ORAC methods). It was generally found that the drying temperature affected the properties of the flour. The flour obtained after drying at 70 ° C (FRCC70) showed higher concentration of active compounds and antioxidant potential in relation to the flour obtained after drying in other temperatures. In this way, the FRCC70 used in the production of cookies. The partial replacement of wheat flour by FRCC70 caused small changes in the physical characteristics (height, diameter, spreading factor and specific volume) of the cookies, mainly for the higher substitution percentages of this flour. In general, cookies with FRCC70 substitution showed higher hardness. It was also verified that the increase of FRCC70 concentration in cookies caused an increase in the concentration of phenolic compounds, likewise, in the antioxidant potential measured by the FRAP and ORAC methods. On the other hand, no effect was observed in the antioxidant potential measured by the DPPH• method. Thus, it can be concluded that the flour of the camu-camu processing byproducts showed potential for application in cookies because it can add nutritional value, due to the increase of the antioxidant potential and the total phenolic concentration of these products.

Scytalidium thermophilum is a termophilic fungus that effectively produces the extracellular enzyme catalase phenol oxidase (CATPO). The enzyme is distinct among catalases with its bifunctionality of oxidising phenolic compounds in the absence of H2O2. CATPO is capable of oxidizing catechol, chlorogenic acid, caffeic acid and catechin which are ortho &ndash / diphenolic compounds. Diphenolic compounds are known as strong antioxidants. Catalase is one of the important antioxidant enzymes. Therefore, in this thesis the effect of CATPO on the final antioxidant capacity of the oxidized products was analysed. Antioxidant capacity measurements of oxidized and unreacted phenolic compounds were done using the two widely used methods TEAC and FRAP. CATPO oxidized catechol showed 2.4 fold increase when compared to its nonoxidized form, which was highest among others. Catechol was followed by caffeic acid, chlorogenic acid, and catechin. This finding is new to the literature and may be of importance to the antioxidant mechanism of organisms. Results have also shown that the most well known phenol oxidases, laccase and tyrosinase, do not result in such high increases in antioxidant capacity upon oxidation of the substrates tested. Due to this finding, as a possible means of applying CATPO to increase the antioxidant capacity of products daily consumed, tea was selected. Tea is the second most consumed beverage after water and it is known to possess high amounts of flavanols. Green tea is rich in catechins whereas black tea is a rich source of theaflavins and thearubigins. Fermentation is a critical process for production of good quality tea and is the key step differing between green and black tea production. During this process phenol oxidases catalyze the oxidation of polyphenolic compounds present in tea leaves to their corresponding o-quinones. Utilization of CATPO in tea samples resulted in an increase in antioxidant capacity and its effect was enhanced by an increase in brewing time. Interestingly, the addition of sugar decreased antioxidant capacity. Laccase and tyrosinase were ineffective in increasing the antioxidant capacity of tea samples.

This study investigated the association between flavonoid profiles of different cowpea (Vigna Unguiculata) varieties with anti-inflammatory properties as a possible benefit against inflammatory bowel disease. Cowpea, a drought tolerant annual herbaceous legume that originated in Africa, is known to possess high levels of polyphenolics, which have demonstrated anti-inflammatory, immunoregulatory and antioxidant properties. Black, red, white, brown and light brown cowpeas were investigated for phenolic content and composition using UV-Visible Spectroscopy and HPLC; antioxidant activation mechanism (AOX) by oxygen radical absorbance capacity (ORAC). Anti-inflammatory activity was measured via NF-κB activation in Raw 264.7 macrophages challenged with a lipo-polysaccharide. Phenols, tannins and AOX activity were generally similar within phenotypes; however among light brown varieties, 09FCV-CC-27M, had among the highest phenols, tannins and AOX, whereas IAR-48 had among the lowest. White cowpea (EARLY ACRE) variety showed the least amount of total phenol content (78.2 mg GAE/g) and condensed tannin content (4.1 mg CE/g); whereas the red varieties (IT82D-889, IT97K-1042-3) contained the highest amounts of tannins (242 and 132 mg CE/g), and phenols (431 and 454 mg GAE/g) respectively. Antioxidant activity correlated with phenol content data. Anthocyanins were only found in the black cowpea. The red varieties had the highest levels of flavonols, which were mostly quercetin derivatives; the white and light brown (IAR-48) varieties had quercetin-3-O-diglucoside as the dominantcompound. The light brown variety (09FCV-CC-27M) had the highest amount of flavan-3-ols while in the white variety no flavan-3-ols were detected. Unexpectedly, the cowpea extracts with lower phenolic and tannins content, the white and light brown (IAR-48) varieties, showed significant (p<0.05) anti-inflammatory properties in the LPS induced macrophages, inhibiting the activation of NF-κB at different concentrations (0.33, 1.67 and 3.33 μg extract/mL). Conversely, extracts with higher phenolic and tannin content did not induce anti-inflammatory response at similar concentrations, suggesting that tannins or other phenolics interfered with anti-inflammatory response at these concentrations. These results suggest that cowpea composition is an important determinant of anti-inflammatory response in inflammatory bowel disease.

No description available.

Verticillium wilt is a serious disease in a wide range of economic crops worldwide. Verticillium wilt of potato is caused, primarily, by the fungus Verticillium dahliae. Disease management requires understanding of V. dahliae pathogenesis and interactions with potato, which was the main objective of this study. A differential potato-V. dahliae pathosystem was established where pathogenicity of four V. dahliae isolates with different levels of aggressiveness was evaluated on two potato cultivars, Kennebec (susceptible) and Ranger Russet (moderately resistant). External and internal symptoms and growth measurements revealed that isolates Vd1396-9 and Vs06-14 are highly and weakly aggressive, respectively. These two isolates were selected for transcriptomics and proteomics investigations to identify pathogenicity-related factors. Transciptomics analysis was conducted in both isolates after elicitation by root extracts from either Kennebec or Ranger Russet using a combinational approach involving subtractive hybridization and cDNA-AFLP. A total of 573 differentially expressed transcripts were detected in one or the other isolate. Among them, 185 transcripts of interest were recovered, re-amplified, sequenced and searched against NCBI and the Broad Institute V. dahliae genome databases for identification. The two contrasting-aggressiveness isolates were used for a comparative proteomics investigation. The first proteomic map of V. dahliae was established. The proteomics analysis was carried out using 2-Dimentional electrophoresis and mass spectrometry. Twenty five proteins were differentially expressed and identified in one or the other isolate. Many of the identified genes/proteins showed potential involvement in pathogenesis of V. dahliae or other fungi. Genes of stress response regulator A (oxidative stress tolerance factor), isochorismatase hydrolase (potential plant defense suppressor) and tetrahydroxynaphthalene reductase (involved in melanin and microsclerotia formation) were isolated from both isolates and cloned. Sequence analysis of these genes showed many differences that may explain their differential expression in the two isolates. Given that some of the identified genes/proteins are potentially involved in overcoming and suppressing plant defense, phenolics were profiled in Kennebec-inoculated with Vd1396-9 or Vs06-14 isolate. Chlorogenic, caffeic, ferulic acids, cis-jasmone and rutin accumulation showed variations after inoculation. The results obtained from this study will help understanding the V. dahliae-potato interactions and develop efficient strategies to control Verticillium wilt disease.

Sinapic acid derivatives and tocopherols in refining byproducts of commercially produced expeller-pressed canola oils were characterized and isolated. Additionally, the antioxidant activities of the phenolics were examined by three systems including an in vitro non-biological related assay, a cellular assay and a deep-fat frying model. Sinapic acid (SA： 42.9 µg/g), Sinapine (SP: 199 µg/g), and Canolol (CAN: 344 µg/g) were found in different byproducts of canola oil refining, namely, soapstock, spent bleaching clay, and wash-water, respectively. Tocopherols (3.75 mg/g) and other non-identified phenolic compounds (2.7 mg /g) were found in deodistillates (DDL). CAN and DDL revealed significant protection effect (p<0.05) against hydrogen peroxide induced oxidation in two mammalian cell lines. The results of deep-fat frying studies indicated positive effects of CAN and DDL in preventing lipid oxidation. The canola oils fortified with DDL and CAN showed a considerable reduction (p < 0.05) in oxidation products of lipid after frying.

Verticillium wilt is a serious disease in a wide range of economic crops worldwide. Verticillium wilt of potato is caused, primarily, by the fungus Verticillium dahliae. Disease management requires understanding of V. dahliae pathogenesis and interactions with potato, which was the main objective of this study. A differential potato-V. dahliae pathosystem was established where pathogenicity of four V. dahliae isolates with different levels of aggressiveness was evaluated on two potato cultivars, Kennebec (susceptible) and Ranger Russet (moderately resistant). External and internal symptoms and growth measurements revealed that isolates Vd1396-9 and Vs06-14 are highly and weakly aggressive, respectively. These two isolates were selected for transcriptomics and proteomics investigations to identify pathogenicity-related factors. Transciptomics analysis was conducted in both isolates after elicitation by root extracts from either Kennebec or Ranger Russet using a combinational approach involving subtractive hybridization and cDNA-AFLP. A total of 573 differentially expressed transcripts were detected in one or the other isolate. Among them, 185 transcripts of interest were recovered, re-amplified, sequenced and searched against NCBI and the Broad Institute V. dahliae genome databases for identification. The two contrasting-aggressiveness isolates were used for a comparative proteomics investigation. The first proteomic map of V. dahliae was established. The proteomics analysis was carried out using 2-Dimentional electrophoresis and mass spectrometry. Twenty five proteins were differentially expressed and identified in one or the other isolate. Many of the identified genes/proteins showed potential involvement in pathogenesis of V. dahliae or other fungi. Genes of stress response regulator A (oxidative stress tolerance factor), isochorismatase hydrolase (potential plant defense suppressor) and tetrahydroxynaphthalene reductase (involved in melanin and microsclerotia formation) were isolated from both isolates and cloned. Sequence analysis of these genes showed many differences that may explain their differential expression in the two isolates. Given that some of the identified genes/proteins are potentially involved in overcoming and suppressing plant defense, phenolics were profiled in Kennebec-inoculated with Vd1396-9 or Vs06-14 isolate. Chlorogenic, caffeic, ferulic acids, cis-jasmone and rutin accumulation showed variations after inoculation. The results obtained from this study will help understanding the V. dahliae-potato interactions and develop efficient strategies to control Verticillium wilt disease.

Grape pomace is a by-product of wineries. It is one of the most potent antioxidant sources due to its high phenolic content. In this thesis study, ultrasound assisted extraction of phenolic compounds from Merlot grape pomace has been studied. The effects of sonication time, subsequent extraction time in shaking water bath at 45&deg / C and composition of the solvent on extraction efficiency and recovery of phenolics were studied by response surface methodology. Folin-Ciocalteu colorimetric method was used to analyze effects of process parameters on the total phenolic content of the extracts. The best recovery (47.2 mg gallic acid equivalents of total phenolics per g of dried grape pomace) was obtained using 30 % aqueous ethanol and applying 6 minutes of sonication followed by 12 minutes of shaking in water bath at 45&deg / C.

The consequences of pre-fermentation and post-fermentation extended maceration (EM) on colour, mouthfeel and phenolic composition were investigated in Pinot noir (2004), Grenache (2006) and Shiraz (2007) (Vitis vinifera L.) wines. Experimental wines were made using cold soak, post-fermentation EM, and standard fermentation treatments (C). Cold soak treatments included a 3-day cold soak at 10°C (CS), with an additional comparison of plunging effects for the 2004 wines (CSP). Post-fermentation extended maceration treatments were 1- or 3-weeks on skins in 2004 (PS1 and PS3 respectively), and 3-weeks in 2006 and 2007 (PS3). A variety of chemical and sensory test methods were used to determine changes in phenolic components and organoleptic properties between treatments of all 3 varietals as they aged in the bottle. Among other results, it was determined if wines made with a period of cold soak had increased colour intensity, and increased concentrations of monomeric anthocyanins and pigmented polymers compared to control wines. It was also determined if wines made with a period of post-fermentation EM had increased concentrations of the flavan-3-ols (+)-catechin and (−)-epicatechin, and tannin, decreased colour intensity and modified mouthfeel compared to the other wine treatments. A greater understanding of Australian red winemakers’ opinions on EM regimes and their use in Australian wineries was obtained by survey. Survey results confirmed that EM is used extensively in Australian wineries but that winemakers have poor understanding of the consequences of EM regimes for red wine properties. The survey confirmed that winemakers are concerned about the economic cost and logistic pressures associated with the use of EM regimes during vintage. Wines made using EM need to spend longer in fermentation vessels, which are in high demand during this time. Findings from this study provide winemakers with more information to consider before making decisions about their use of EM regimes. Survey findings showed more winemakers would use EM regimes if logistic and economic pressures did not apply. However, results suggest that even if winemakers did adopt EM practices, some may not achieve what they believe to be the outcome of these regimes, such as improved colour or mouthfeel properties. For instance, results showed that cold soaking did not make a difference to wine colour compared to conventional fermentation maceration. Even without cold soaking red must, winemakers may be able to achieve the same or very similar wine organoleptic characteristics at a reduced cost. Similarly, no significant effects of plunging during cold soak were observed. Post-fermentation EM visibly reduced wine colour intensity and imparted a browner hue to the wine compared to red wine that was pressed off skins upon reaching dryness. This EM regime is therefore unlikely to benefit winemakers who are seeking to produce highly coloured wines. However, prolonged maceration post-fermentation did increase the intensity of perceived bitterness and increased the concentration of wine flavan-3-ols and tannins. Winemakers may therefore influence the desired balance between the extraction of these wine phenolics (and the associated outcome for taste and mouthfeel properties) and economic considerations by varying the duration of maceration post-fermentation. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1362928 / Thesis (Ph.D.) - University of Adelaide, School of Agriculture, Food and Wine, 2009.

The antioxidant capacity of phenolic compounds in red wine is suggested to be responsible for their health-promoting effects. Compared to other wines, little information is available on phenolic compositions and antioxidant capacity of Australian wine. Information related to the fate of these phenolics in the body once consumed is also very limited. The overall aim of this research was to investigate the relevance of red wine consumption as a source of health-giving antioxidants in humans. The phenolic composition of wine was determined using the Folin-Ciocalteu (total phenolic), aluminium chloride (total flavonols), methyl cellulose precipitation (MCP) (total tannins), pH differential (total monomeric anthocyanins), bisulfite bleaching (total polymeric anthocyanin fractions), and liquid chromatographic (LC-MS) (individual phenolics) methods. Antioxidant activities were measured using 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2’-azino-bis-93-ethylbenzthiazoline-6-sulfonic acid (ABTS) and oxygen radical absorbance capacity (ORAC) assays. The phenolic and antioxidant data were then used to establish the relationship between these two parameters in wines from different varieties (Shiraz, Cabernet Sauvignon and Merlot) and winemaking stages (crushing, fermentations, oaking and bottling). By using an in vitro digestion model that mimics the upper gastrointestinal tract (GIT) digestion, the stability of the wine phenolics during digestion was examined. Finally, to gain a better understanding of the post-digestion absorption of wine phenolics, their permeability across Caco-2 cell monolayers was evaluated. A total of 8 monomeric anthocyanins and 17 other phenolic compounds were positively identified in the red wines using LC-MS analysis. Most of the phenolic categories showed some positive correlations with the antioxidant activities but none of the individual phenolic compounds showed a strong correlation with the total antioxidant activity of the wine, implying a combined contribution of many wine phenolics to antioxidant effects. The phenolic compositions and antioxidant activities of three of Australia’s most common red wines varieties - Shiraz, Cabernet Sauvignon and Merlot were not different from each other, possibly due to the variability within each grape cultivar. During the winemaking process, the total phenolic content and the associated antioxidant activity of the wine increased during the fermentation process, as more phenolics are being extracted from grape skin, seeds and stems into the wine. During oak and bottle ageing, the total phenolic contents were stabilised. Most of the wine phenolics were more stable under acidic conditions (pH 2 and 5.5) than neutral or alkaline conditions (pH 7.4 and 9). This may partly explain the stability of the wine phenolics subjected to the acidic (pH 2) gastric digestion and their loss following simulated pancreatic digestion (pH 7.4). In addition, sample pre-treatment procedures prior to LC-MS analysis may have removed some antioxidants in the form of degradation products and/or new polymeric compounds following the in vitro gastric and pancreatic digestion processes. The missing products appeared to be detected by both the Folin-Ciocalteu method and ORAC assay, which measured the phenolic compounds and their antioxidant activity, after the pancreatic digestion. This suggests that the instability of phenolic compounds at pH 7.4, results in the transformation of most of the oral phenolic antioxidants into more stable forms in the GIT, which in turn contribute positively to the overall antioxidant activities of the ingested wine. All of the original wine phenolics had very low permeabilities across Caco-2 cell monolayers, except for syringic acid, p-coumaric acid and an unknown phenolic acid. Limited surface area for absorption (0.33 cm2) and the limited peak detection sensitivity in the LC method may have contributed towards the difficulty in detecting and identifying compounds with low permeability. In addition, extensive metabolism of absorbed phenolics by the Caco-2 cells may occur based on the appearance of several new peaks. However, due to their low concentrations and lack of reference, the identities of the new products and metabolites remain unknown. The present in vitro study suggests that upon ingestion, most of the original phenolic compounds in red wine are lost either through degradation to new compounds and/or complexation with other compounds. However, these products seem to possess some antioxidant activity and may be the key compounds responsible for the health-promoting effects of red wine. The limitation of the present study in detecting and fully identifying these breakdown products and metabolites should be addressed in future studies.