Hydromonas duriensis gen. nov., sp. nov., isolated from freshwater.

An aerobic, Gram-stain-negative rod, designated strain A2P5T, was isolated from the Douro river, in Porto, Portugal. Cells were catalase- and oxidase-positive. Growth occurred at 15-30 °C, at pH 6-8 and in the presence of 1 % (w/v) NaCl. The major respiratory quinone was Q8, the genomic DNA had a G+C content of 47 ± 1 mol%, and phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol were amongst the major polar lipids. On the basis of 16S rRNA gene sequence analysis, strain A2P5T was observed to be a member of the family Burkholderiaceae, but could not be identified as a member of any validly named genus. The low levels of 16S rRNA gene sequence similarity to other recognized taxa ( < 91 %), together with the comparative analysis of phenotypic and chemotaxonomic characteristics, supported the proposal of a novel species of a new genus within the family Burkholderiaceae. The name Hydromonas duriensis gen. nov., sp. nov. is proposed. The type strain of Hydromonas duriensis is A2P5T ( = LMG 28428T = CCUG 66137T).

Discrimination of white wine ageing based on untarget peak picking approach with multi-class target coupled with machine learning algorithms.

The complexity of the chemical reactions occurring during white wine storage, such as oxidation turns the capacity of prediction and consequently the capacity to avoid it extremely difficult. This study proposes an untarget methodology based on machine learning algorithms capable to classify wines according to their "oxidative-status". Instead of the most common approach in statistics using one class for classification, in this work eight classes were selected based on target oxidation markers for the extraction of relevant compounds. VIPS from OPLS-DA and mean decrease accuracy from random forest were used as feature selection parameters. Fifty-one molecules correlated with 5 classes, from which 23 were selected has having higher sensitivities (AUC > 0.85). For the first time to our knowledge hydroxy esters ethyl-2-hydroxy-3-methylbutanal and ethyl-2-hydroxy-4-methylpentanal were found to be correlated with oxidation markers and consequently to be discriminant of the wine oxidative status.

Impact of Phenolic Compounds in Strecker Aldehyde Formation in Wine Model Systems: Target and Untargeted Analysis.

The Strecker degradation of phenylalanine has been studied in a phenolic compound/phenylalanine wine model system. Six phenolic compounds (3,4-dihydroxybenzoic acid, gallic acid, caffeic acid, ferulic acid, catechin, and epicatechin) were compared in the formation of phenylacetaldehyde when in the presence of glucose or methylglyoxal (MG). The addition of glucose reduced the formation of Strecker aldehyde, independently of the phenolic compound. The addition of MG, on the other hand, increased phenylacetaldehyde formation for hydroxybenzoic acids and decreased phenylacetaldehyde formation for flavan-3-ols, confirming their capacity to trap the dicarbonyl compound. As a target phenolic compound, catechin was chosen to perform kinetic studies to further understand the reaction intermediates involved in the mechanism of phenylacetaldehyde formation, in particular, catechin o-quinone and catechin-MG adduct. The addition of glucose and MG increased the consumption of catechin, while a reduction in the respective o-quinone was observed, suggesting that these substrates have an impact in other reactions involving catechin. In that regard, for the first time, it was demonstrated that the catechin-MG adduct was capable of oxidizing and forming a new o-quinone, contributing to wine instability promoted by oxidation reactions.

Resistance to oxidation of white wines assessed by voltammetric means.

This work concerns the development of a methodology suited to measure the resistance to oxidation of white wines by cyclic voltammetry. The voltammetric responses of several white wines of different origin and age were analyzed in the oxidation potential range (0.2-1.2 V vs SCE). Currents measured at fixed potentials were correlated to the concentration of ascorbic acid, SO2, and total phenolics. A forced degradation study was monitored by cyclic voltammetry; from plots of current versus time, the consumption rates of oxidizable species in wine were estimated.

Chemiomics: network reconstruction and kinetics of port wine aging.

Network reconstruction (NR) has proven to be useful in the detection and visualization of relationships among the compounds present in a Port wine aging data set. This view of the data provides a considerable amount of information with which to understand the kinetic contexts of the molecules represented by peaks in each chromatogram. The aim of this study was to use NR together with the determination of kinetic parameters to extract more information about the mechanisms involved in Port wine aging. The volatile compounds present in samples of Port wines spanning 128 years in age were measured with the use of GC-MS. After chromatogram alignment, a peak matrix was created, and all peak vectors were compared to one another to determine their Pearson correlations over time. A correlation network was created and filtered on the basis of the resulting correlation values. Some nodes in the network were further studied in experiments on Port wines stored under different conditions of oxygen and temperature in order to determine their kinetic parameters. The resulting network can be divided into three main branches. The first branch is related to compounds that do not directly correlate to age, the second branch contains compounds affected by temperature, and the third branch contains compounds associated with oxygen. Compounds clustered in the same branch of the network have similar expression patterns over time as well as the same kinetic order, thus are likely to be dependent on the same technological parameters. Network construction and visualization provides more information with which to understand the probable kinetic contexts of the molecules represented by peaks in each chromatogram. The approach described here is a powerful tool for the study of mechanisms and kinetics in complex systems and should aid in the understanding and monitoring of wine quality.

3-Hydroxy-4,5-dimethyl-2(5H)-furanone: a key odorant of the typical aroma of oxidative aged Port wine.

Application of aroma extract dilution analysis (AEDA) on organic extracts from Port wines barrel-aged over 40 years revealed 5 odor-active compounds corresponding to descriptors used to qualify the characteristic old wine aroma. One of the compounds, described as "nutty" and "spicy-like", and present in at least 9 dilutions above the others, was perceived as particularly important. The compound responsible for this flavor was identified as 3-hydroxy-4,5-dimethyl-2(5H)-furanone (sotolon). The levels ranged from 5 to 958 microg/L for wines between 1 and 60 years old. It was also observed that during oxidative aging the concentration of this compound increased with time according to a linear trend (r > 0.95). Although the presence of 2-ketobutyric acid was verified, the constant rate of formation of sotolon with aging and its high correlation with sugar derivates (HMF, furfural) suggests other mechanisms, different from those reported for other wines. The flavor threshold of sotolon was evaluated in Port wine at 19 microg/L. Sensorial tests provided valuable information concerning sotolon impact on Port wine aroma. Samples supplemented with this substance were consistently ranked as older. In view of these results it can be expected that sotolon plays a pre-eminent role in the characteristic old Port wine aroma.

Relationship between potentiometric measurements, sensorial analysis, and some substances responsible for aroma degradation of white wines.

Oxidative degradation of white wines can be described sensorially as developing from a loss at positive aroma characteristics, through the development of negative aromas to a linel stage of chromatic alterations. This work attempts to relate the oxidation "status" evaluate by potentiometric titrations, with sensorial degradation and the levels of substances responsible for "off-flavors", such as methional and phenylacetaldehyde. The potentiometric titration employed measures the most powerful antioxidants of white wines (e.g., those which more rapidly consume oxygen). Considering that aromatic precedes chromatic degradation, resistance to oxidation (ROX) constitutes a useful indicator of resistance to oxidation. Sensorial degradation (ID), potentiometric measures, and volatiles were determined both in samples submitted to a "forced aging" protocol and normal aged white wines. High correlation values were observed between ROX and the ID, in both sets (r > 0.87). ID is better explained by ROX values than by the indicated wine age or by the "degree of browning" (Abs = 420 nm). It was also observed that in samples with ROX values higher than 10, the concentration of methional and phenylacetaldehyde were above their respective odor threshold. Finally, it was observed that there is a relationship between oxygen consumption and the respective ROX. Although these results seem very promising, they needed to be further complemented in order to estimate the shelf life of a white wine using potentiometric titrations.

Further insights into the role of methional and phenylacetaldehyde in lager beer flavor stability.

This work attempts to measure the importance of methional and phenylacetaldehyde on the flavor stability of beer, as direct participants or as indicators of aroma deterioration. A trained sensory panel identified the most important descriptors related to the typical aroma of aged beer: "malty", "honey-like", "cooked potato", and "metallic". By GC-olfactometry analysis, six aromatic zones related to the selected descriptors were highlighted, and by using GC-MS techniques it was possible to identify methional and phenylacetaldehyde as being responsible for two odor zones. The quantification of these molecules in samples submitted to forced aging treatments showed that the levels of methional and phenylacetaldehyde are dependent on the temperature of storage. Normal aged beers were also analyzed, and it was observed that these compounds accumulate with time of storage. Furthermore, these molecules were negatively correlated with the aroma quality of beer as evaluated by a sensorial panel. To validate the sensory impact of these substances, a fresh beer was spiked with these molecules and also with trans-2-nonenal, singly and in combination, and the similarity value between samples and the aged beer was then determined. The highest value from the similarity tests was 72% when the three compounds were added simultaneously. The combination of the two Strecker aldehydes increases by 54% the degree of similarity, indicating the key role played by these molecules in the aroma deterioration of beer. Finally, the kinetic parameters, Ea and k, were calculated, and it was observed that the Arrhenius equation described well the temperature dependence of the reaction rate constant. Measuring the concentration of methional and phenylacetaldehyde may provide information about the key steps along the process that most affect the flavor stability of beer, which may be useful in establishing the best storage conditions.

Carotenoid compounds in grapes and their relationship to plant water status.

The aim of this work was to study the relationship between carotenoid contents in grapevine berries and plant water status. For this purpose, a black grapevine variety, Vitis vinifera L. cv. Touriga Nacional, was studied. The experiments were carried out in the same Douro vineyards, with plants of the same age, in two different water retention soils. A higher water retention capacity soil, soil A, and a lower water retention capacity soil, soil B, were both in a 1.2 m deep silt-loam schist-derived soil. The training system was the double cordon trained and spur pruned. A first range was nonirrigated (NI) and a second one was irrigated (I), 60% of evapotranspiration (ET(0)). For soil B, a 30% of ET(0) treatment was also applied. The plant water status was estimated by predawn leaf water potential. The effects of plant water status on berry growth were studied by measurement of the berry weight and total soluble solids (degrees Brix). The carotenoid profile was quantitatively determined by high-performance liquid chromatography/diode array. Carotenoids determined were beta-carotene, lutein, neoxanthin, violaxanthin, and luteoxanthin. The comparison between irrigated and nonirrigated grapes was followed from 2 weeks before veraison until the ripe stage. Results showed that at harvest time, berries exposed to the NI had a lower weight than those exposed to the irrigated treatment (60% of ET(0)), 0.89 vs 1.36 g/berry and 0.94 vs 1.34 g/berry, for soils A and B, respectively. The irrigated treatment contributed to a higher sugar concentration in both soils. However, depending on the soil water retention capacity, the carotenoid contents were different in soils A and B. For soil A, the total carotenoid content was similar for both NI and I treatments. However, with regard to soil B, in irrigated treatment, levels of carotenoids were approximately 60% lower than those found for the NI. It seems to be possible to produce higher weight berries (with higher sugar levels) with similar carotenoid contents. On the other hand, soil characteristics had a larger influence than irrigation on the concentration of carotenoids in grapes, resulting in an important viticultural parameter to take into account in aroma precursor formation.

Determination of carotenoid profiles in grapes, musts, and fortified wines from Douro varieties of Vitis vinifera.

beta-Carotene and six xanthophylls (lutein, neoxanthin, violaxanthin, luteoxanthin, cryptoxanthin, and echinenone) have been identified and semiquantitatively or quantitatively determined in musts and port wines for the first time. An HPLC method was developed and compared with that of one based on thin layer cromatography with scanning densitometry. The most abundant carotenoids present in red grape varieties are beta-carotene and lutein. In wines, significant quantities of violaxanthin, luteoxanthin, and neoxanthin were found. This study was done with berries (skin and pulp), musts, and fortified wines. Some experiments were performed to follow carotenoid content from grapes to wines. Although the levels of beta-carotene and lutein found in fortified wines were lower than those found in musts, other xanthophylls, such as neoxanthin, violaxanthin, and luteoxanthin, exist in appreciable amounts in young ports.

Volatile free fatty acids as ripening indicators for Serra da Estrela cheese.

Serra da Estrela cheese is an artisanal cheese manufactured from raw ewe's milk coagulated with Cynara cardunculus; it is highly appreciated for its unique flavor and bouquet. This research effort focused on a search for the molecules responsible for those organoleptic characteristics. Eighty cheeses manufactured in 4 dairies located in the Appelation d'Origine Protegee region of Serra da Estrela, Portugal, were thus assayed for volatiles, in an attempt to characterize their odorous profile. Cheeses were analyzed from the time of manufacture up to 180 d of ripening. The volatile fraction was assayed by solid phase microextraction-gas chromatography/mass spectroscopy, and several compounds belonging to different chemical groups (e.g., fatty acids, esters, carbonyl compounds, pyrazines, and sulfur compounds) were detected in that traditional cheese. Among these, free fatty acids (FFA) were quantitatively the dominant family present. Furthermore, sensory descriptors for the typical aroma of this cheese included acidic, sweaty, and sheepy-like. Acetic, isobutyric, and isovaleric acids increased in concentration during the ripening process up to 90 d, and remained constant thereafter. Preliminary sensory analysis was performed by addition of the major FFA to an unripened cheese matrix; results showed that they could successfully be used as ripening indicators for this cheese. Such key molecules may thus be used to monitor ripening, and hence find the optimal consumption time for this gourmet dairy product.