Analytical and sensorial characterization of the aroma of wines produced with sour rotten grapes using GC-O and GC-MS: identification of key aroma compounds

In the present work, the aroma profiles of wines elaborated from sound and sour rot-infected grapes as raw material have been studied by sensory analysis, gas chromatography-olfactometry (GC-O), and gas chromatography-mass spectrometry (GC-MS), with the aim of determining the odor volatiles most likely associated with this disease. The effect of sour rot was tested in monovarietal wines produced with the Portuguese red grape variety Trincadeira and in blends of Cabernet Sauvignon and sour rotten Trincadeira grapes. Wines produced from damaged berries exhibited clear honey-like notes not evoked by healthy samples. Ethyl phenylacetate (EPhA) and phenylacetic acid (PAA), both exhibiting sweet honey-like aromas, emerged as key aroma compounds of sour rotten wines. Their levels were 1 order of magnitude above those found in controls and reached 304 and 1668 μg L(-1) of EPhA and PAA, respectively, well above the corresponding odor thresholds. Levels of γ-nonalactone also increased by a factor 3 in sour rot samples. Results also suggest that sour rot exerts a great effect on the secondary metabolism of yeast, decreasing the levels of volatiles related to fatty acids and amino acid synthesis. The highest levels of γ-decalactone of up to 405 μg L(-1) were also found in all of the samples, suggesting that this could be a relevant aroma compound in Trincadeira wine aroma.     

Heterocyclic acetals from glycerol and acetaldehyde in Port wines: evolution with aging

In Port wine, isomers of glycerol and acetaldehyde acetals have been found at total contents ranging from 9.4 to 175.3 mg/L. During oxidative aging, the concentrations of the 5-hydroxy-2-methyl-1,3-dioxane and 4-hydroxymethyl-2-methyl-1,3-dioxolane isomers increased with time showing a linear correlation (r > 0.95). The flavor threshold for the mixture of the four isomers was evaluated in wine at 100 mg/L. Thus, it is expected that they contribute to "old Port wine" aroma in wines older than 30 years. Experiments with model solutions and wine clearly demonstrated that SO(2) combines with acetaldehyde and blocks the acetalization reaction.     

Gas chromatography-olfactometry and chemical quantitative study of the aroma of six premium quality spanish aged red wines

The aroma of six premium quality Spanish red wines has been studied by quantitative gas chromatography-olfactometry (GC-O) and techniques of quantitative chemical analysis. The GC-O study revealed the presence of 85 aromatic notes in which 78 odorants were identified, two of which-1-nonen-3-one (temptatively) and 2-acetylpyrazine-are reported in wine for the first time. Forty out of the 82 quantified odorants may be present at concentrations above their odor threshold. The components with the greatest capacity to introduce differences between these wines are ethyl phenols produced by Brettanomyces yeasts (4-ethylphenol, 4-ethyl-2-methoxyphenol, and 4-propyl-2-methoxyphenol), 2,5-dimethyl-4-hydroxy-3(2H)-furanone (furaneol), (Z)-3-hexenol, thiols derived from cysteinic precursors (4-methyl-4-mercaptopentan-2-one, 3-mercaptohexyl acetate, and 3-mercaptohexanol), some components yielded by the wood [(E)-isoeugenol, 4-allyl-2-methoxyphenol, vanillin, 2-methoxyphenol (guaiacol), and (Z)-whiskylactone], and compounds related to the metabolism (2-phenylethanol, ethyl esters of isoacids, 3-methylbutyl acetate) or oxidative degradation of amino acids [phenylacetaldehyde and 4,5-dimethyl-3-hydroxy-2(5H)-furanone (sotolon)]. The correlation between the olfactometric intensities and the quantitative data is, in general, satisfactory if olfactometric differences between the samples are high. However, GC-O fails in detecting quantitative differences in those cases in which the olfactive intensity is very high or if odors elute in areas in which the odor chromatogram is too complex.     

Aroma extract dilution analysis. Precision and optimal experimental design

The odor thresholds of 12 different compounds have been determined in an AEDA experiment using a panel composed of 8 individuals. Only in one case is the distribution of thresholds among judges significantly different from the log-normal. The cause of that departure from normality seems to be a cross adaptation rather than anosmia. The standard deviations (SD) range from 2(0.7) to 2(4.1), with 2(1.8) as average. If the AEDA is carried out at a dilution rate, R, and dilution R(P) (where P = 0, 1, 2.) is the last one at which a compound was detected by a judge, the flavor dilution (FD) factor that should be provided for that judge is R(P+0.5). In the case where several judges carry out the AEDA, the FD for a given compound should be the geometric mean of the FD provided by all the judges. The SD of the distribution of FDs is related to that of the distribution of odor thresholds, but is strongly influenced by the dilution rate, R, being higher with higher R values. The relationship between both SDs can be used to determine the expected precision (measured as the geometric length of the 95% confidence interval) of the result of an AEDA experiment as a function of the dilution rate, the number of judges, and the SD of the distribution of thresholds. Different simulations have shown that in most cases, a dilution rate of 10 is the best option, and that lower dilution rates are advantageous only if the analyzed solution contains compounds with a very narrow distribution of thresholds.     

Role of Azospirillum brasilense nitrate reductase in nitrate assimilation by wheat plants

Summary The nitrogen metabolism of wheat plants inoculated with various Azospirillum brasilense strains and nitrate reductase negative (NR^–) mutants was studied in two monoxenic test tube experiments. The spontaneous mutants selected with chlorate under anaerobic conditions with nitrite as terminal electron acceptor fixed N₂ in the presence of 10 mM NO₃ ^– and were stable after the plant passage. One strain (Sp 245) isolated from surface-sterilized wheat roots produced significant increases in plant weight at both NO₃ levels (1 and 10 mM) which were not observed with the NR^– mutants or with the two other strains. Similar effects were observed in a pot experiment with soil on dry weight and total N incorporation but only at the higher N fertilizer level. In the monoxenic test tube experiments plants inoculated with the mutants showed lower nitrogenase activities than NR⁺ strains at the low NO₃ ^– level (1 = mM) but maintained the same level of activity with 10 mM NO₃ ^– where the activity of all NR⁺ strains was completely repressed. The nitrate reductase activity of roots increased with the inoculation of the homologous strains and with the mutants at both NO₃ ^– levels. At the low NO₃ ^– level this also resulted in increased activity in the shoots, but at the high NO₃ ^– level the two homologous strains produced significantly lower nitrate reductase activity in shoots while the mutants more than doubled it. The possible role of the bacterial nitrate reductase in NO₃ ^– assimilation by the wheat plant is discussed.     

Heavy metal and PCB spatial distribution pattern in sediments within an urban catchment—contribution of historical pollution sources

Purpose

This work analyzes polychlorinated biphenyl (PCB) and heavy metal contamination in fluvial sediments and soils in an urban catchment, according to the geo-accumulation index and to soil and sediment quality guidelines. The catchment is located in Coimbra, Portugal, being affected by frequent flooding, and its main stream is a tributary to one of the major rivers in Portugal (Mondego). Given the presence of industrial activities over time, some inputs of pollutants are expected, but so far, the legacy of historic pollution in this catchment has not yet been investigated.

Materials and methods

Twenty-five samples were collected from nine sampling sites at the depths of 0–20, 20–40 and 40–60 cm (to provide a historic perspective) along longitudinal profiles (streamlines) and in soils downstream of pollution sources. These samples were analyzed for six heavy metals (Cu, Cr, Pb, Cd, Zn, Ni), organic carbon, pH and ten PCBs (IUPAC numbers 28, 30, 52, 101, 138, 153, 166, 180, 204, 209).

Results and discussion

Total PCB concentrations ranged 0.47–5.3 ng g^?1 dry weight (dw), and levels increase from the bottom to the top layers, suggesting an increased input over the last 100 years. PCB congener distribution shows the dominance of hexachlorobiphenyls, especially PCB138, suggesting the existence of local sources. PCB levels did not exceed sediment quality levels, placing sediments/soil under class 1 (not contaminated) or class 2 (trace contamination) with respect to PCB. All six metals exceeded the lowest effect level for sediment quality criteria, and three (Cd, Pb and Zn) largely exceeded the clean levels for dredged materials, placing sediments in class 5 (heavily contaminated). Sampling site S1 presented the highest concentrations of Zn, Pb and Cd, and historic vehicle traffic was identified as the most likely source, given the vertical and horizontal profiles.

Conclusions

High levels of Pb, Cd and Zn were found in fluvial sediments at some locations of the Loreto catchment, likely from historic traffic sources. This urban area is frequently affected by flooding events and is currently being subject to urban redevelopment. During these events/actions, historic pollutants in the sediments might surface and be redistributed, impacting the downstream ecosystem of the major Mondego River or increasing the risk of exposure of the urban population.

     

Leukocyte-hepatocyte interaction in calicivirus infection: differences between rabbits that are resistant or susceptible to rabbit haemorrhagic disease (RHD)

Calicivirus infection is lethal for adult rabbits, whereas young rabbits (less than 8-weeks-old) are resistant to the same infectious agent. The virus replicates in the liver and causes a fulminant hepatitis in adult rabbits leading to rabbit haemorrhagic disease (RHD); this is in contrast with the mild and transient hepatitis observed in infected young rabbits. We have used electron microscopy to compare liver leukocyte infiltrates between young (resistant) and adult (susceptible) rabbits, 36-48 h after inoculation of the animals with caliciviruses. In adult rabbits, liver infiltrates were made up mostly of heterophils, and they were located near hepatocytes showing severe cellular damage. In contrast, liver leukocyte infiltrates of RHD-resistant young rabbits were dominated by lymphocytes that depicted membrane contacts with the cell surface of undamaged hepatocytes. We conclude that: (i) the cellular inflammatory response of the liver to calicivirus infection is different in rabbits that are susceptible (adult) or resistant (young) to RHD; (ii) leukocyte infiltration of the adult liver by heterophils is probably directed at the removal of dead hepatocytes, whereas the liver lymphocytic infiltration of young rabbits suggests the expression of viral antigens on the surface of liver cells of the RHD-resistant animals.     

Physiological reactions of three breeds of goats to cold,heat and high altitude

A total of 36 goats of the breeds Gemsfarbige Gebirgsziege (G), Saanen (S) and Toggenburger (T) were exposed for periods of 4 hours to the following climatic conditions: normal (20°C, 50% RH); cold (?5°C); heat (40°C, 30% RH) and high altitude (4000 m, 20°C, 50% RH).In the cold the animals were just able to maintain normal core temperature. Skin temperatures on the flank decreased by 3–5°C, those on the ear and the shank by 20–21°C. Respiratory rate and blood-pH decreased. The elevation in heart rate (18 beats/min) and cold shivering were indicative of a rise in heat production.In the heat, rectal temperature increased but attained a plateau at 40.7°C. Respiratory rate rose from 26 to 261 breaths/min, producing a slight respiratory alcolosis with a rise in blood-pH of 0.036 units. Heart rate augmented by 15 beats/min.At high altitude rectal temperature declined by 0.2°C, which was attributed to the quiet behaviour of the goats. There were increases in haemoglobin (+0.83 g/100 ml), haematocrit (+2.5 vol.%), erythrocyte number (+0.78 mill./mm³) and specific gravity of the blood (+0.002). The increases were considered to reflect discharge of blood from reservoirs, since they occurred within 4 hours. A rise in pH, unaccompanied by a rise in respiratory rate, pointed to an increase in tidal volume.There were suggestions of breed differences. The T-breed appeared to be relatively cold tolerant owing to the high thermal insulation of their hair coat, and also relatively heat tolerant, possibly due to their low heat production, as evidenced indirectly by their low normal heart rate (87 compared with 95 beats/min in the other two breeds).