Статті в журналах з теми "Fermentation of grape juices with high sugar concentration"

In the tequila industry, fermentation is traditionally achieved at sugar concentrations ranging from 50 to 100 g·L–1. In this work, the behaviour of the Saccharomyces cerevisiae yeast (isolated from the juices of the Agave tequilana Weber blue variety) during the agave juice fermentation is compared at different sugar concentrations to determine if it is feasible for the industry to run fermentation at higher sugar concentrations. Fermentation efficiency is shown to be higher (above 90%) at a high concentration of initial sugar (170 g·L–1) when an additional source of nitrogen (a mixture of amino acids and ammonium sulphate, different than a grape must nitrogen composition) is added during the exponential growth phase.Key words: Saccharomyces cerevisiae, fermentation efficiency, nitrogen source, tequila.

Starmerella bacillaris (synonym Candida zemplinina) is an important non-Saccharomyces yeast in winemaking with valuable oenological properties, accompanying Saccharomyces species in sweet wine fermentation, and has also been suggested for application as combined starter culture in dry or sweet wines. In this study, the major metabolites and nitrogen utilization of these yeasts are evaluated in the musts with high or extremely high sugar concentration. The change in the metabolic footprint of Saccharomyces cerevisiae, Saccharomyces uvarum and Starmerella bacillaris strains was compared when they were present as pure cultures in chemically defined grape juice medium with 220 and 320 g/L of sugar, to represent a fully matured and an overripe grape. Surprisingly, the extreme sugar concentration did not result in a considerable change in the rate of sugar consumption; only a shift of the sugar consumption curves could be noticed for all species, especially for Starmerella bacillaris. At the extreme sugar concentration, Starmerella bacillaris showed excellent glycerol production, moderate nitrogen demand together with a noticeable proline utilisation. The change in the overall metabolite pattern of Starmerella bacillaris allowed clear discrimination from the change of the Saccharomyces species. In this experiment, the adequacy of this non-Saccharomyces yeast for co-fermentation in juices with high sugar concentration is highlighted. Moreover, the results suggest that Starmerella bacillaris has a more active adaptation mechanism to extremely high sugar concentration.

In Russia, fruit wines produced using existing technologies do not yet fully compete with grape wines, as they are inferior to them in terms of organoleptic characteristics and storage stability. In connection with the expansion of the production base of fruit wines and ciders, it is necessary to intensify the study of the chemical composition of local varieties of apples, its changes that occur during alcohol fermentation, as well as throughout the entire technological process for the production of fruit wine, in order to obtain a high-quality product. We studied apple varieties Idared, Golden Delicious, Jonathan, Interprise, Florina, Renet Simirenko, Kore, growing in the Krasnodar Territory, and wines made from them. It has been established that the qualitative composition and concentration of sugars, as well as the organic acids of apple juices, is determined primarily by the varietal characteristics of apples. During the fermentation process, the amount of titrated acids changed. The concentration of malic acid decreased, and lactic acid increased markedly, which led to a softening of the taste of fermented juice and an improvement in its organoleptic characteristics. In apple juices, various groups of phenolic compounds have been identified. The highest concentration of anthocyanins, tannins and catechins was detected in apple varieties with colored skin - Jonathan and Idared. During fermentation, the concentration of phenolic compounds decreased in all fermented juices in comparison with fresh ones. It can be concluded that during fermentation, the main role in biochemical processes is played not only by the varietal characteristics of apples, but also by the physiological properties of the yeast race.

Most commercialized red wines are produced through alcoholic fermentation performed by yeasts of the Saccharomyces genus, and a second fermentation performed by lactic bacteria of the Oenococus oeni species once the first is completely finished. However, the classical process can suffer complications, of which the risks can increase in grape juices with high contents of sugar and pH. Due to climate change, these situations are becoming more common in the winemaking industry. The main risks in those scenarios are alcoholic-fermentation stops or sluggish and undesirable bacteria development while alcoholic fermentation is not finished yet and wine still contains residual sugars. The study propose a novel alternative that offers a solution or reduces the risk of those scenarios while increasing acidity, which is another serious problem of warm viticulture regions. The alternative consists of the combined use of Lachancea thermotolerans to reduce the pH of musts that suffer from a lack of acidity, Lactiplantibacillus plantarum (formerly Lactobacillus plantarum) to achieve malic acid stability during the first stages of alcoholic fermentation, and Saccharomyces bayanus to complete the alcoholic fermentation in difficult wines of high potential alcohol degree of over 15% (v/v). The new proposed biotechnology produced wines with higher final concentrations in lactic acid, glycerol, color intensity, ethyl lactate and 2-phenyl ethyl acetate in 2.39 g/L, 0.52 g/L, 21%, 48% and 37% respectively than the classical methodology where Saccharomyces genus performs alcoholic fermentation and later Oenococus oeni performs malolactic fermentation. Additionally, the new alternative produced wines with lower concentration in ethanol, pH, acetic acid, ethyl acetate, diacetyl and 1-propanol in 0.37% (v/v), 0.26, 0.08 g/L, 22%, 69% and 28% respectively than the classic method.

The effects of Metschnikowia pulcherrima and high glucose osmolality on S. cerevisiae pyruvate dehydrogenase pathway (PDH) by-pass were examined by varying the starting sugar concentration of synthetic grape juice and the inoculation ratio of S. cerevisiae to M. pulcherrima. The findings revealed that M. pulcherrima and osmolarity impacted S. cerevisiae’s PDH by-pass. The inoculation concentration of M. pulcherrima significantly affected pyruvate decarboxylase (PDC) activity and acs2 expression when the initial sugar concentration was 200 g L−1 and 290 g L−1. The osmolarity caused by the initial sugar (380 g L−1) significantly influenced the enzymatic activity of S. cerevisiae, which decreased PDC and acetaldehyde dehydrogenase (ALD) activities while increasing Acetyl-CoA synthetase (ACS) activity. The reduction in acetic acid in the wine was caused by M. pulcherrima altering the initial sugar concentration faced by S. cerevisiae, which in turn affected enzymatic activity. The alteration of enzyme activity and accumulation of primary metabolites revealed why mixed fermentation could reduce the acetic acid content in wine by altering the enzymatic activity and affecting the expression of several key genes. The M. pulcherrima inoculation levels had no significant effect on the acetic acid and glycerol concentration in the same fermentation medium.

ABSTRACTSaccharomyces cerevisiaehas evolved a highly efficient strategy for energy generation which maximizes ATP energy production from sugar. This adaptation enables efficient energy generation under anaerobic conditions and limits competition from other microorganisms by producing toxic metabolites, such as ethanol and CO2. Yeast fermentative and flavor capacity forms the biotechnological basis of a wide range of alcohol-containing beverages. Largely as a result of consumer demand for improved flavor, the alcohol content of some beverages like wine has increased. However, a global trend has recently emerged toward lowering the ethanol content of alcoholic beverages. One option for decreasing ethanol concentration is to use yeast strains able to divert some carbon away from ethanol production. In the case of wine, we have generated and evaluated a large number of gene modifications that were predicted, or known, to impact ethanol formation. Using the same yeast genetic background, 41 modifications were assessed. Enhancing glycerol production by increasing expression of the glyceraldehyde-3-phosphate dehydrogenase gene,GPD1, was the most efficient strategy to lower ethanol concentration. However, additional modifications were needed to avoid negatively affecting wine quality. Two strains carrying several stable, chromosomally integrated modifications showed significantly lower ethanol production in fermenting grape juice. Strain AWRI2531 was able to decrease ethanol concentrations from 15.6% (vol/vol) to 13.2% (vol/vol), whereas AWRI2532 lowered ethanol content from 15.6% (vol/vol) to 12% (vol/vol) in both Chardonnay and Cabernet Sauvignon juices. Both strains, however, produced high concentrations of acetaldehyde and acetoin, which negatively affect wine flavor. Further modifications of these strains allowed reduction of these metabolites.

The aim of this work was to study the production of kefir-like beverages via the fed-batch fermentation of red table grape juice at initial pHs of 3.99 (fermentation A) and 5.99 (fermentation B) with kefir grains during 4 repeated 24-h fed-batch subcultures. All kefir-like beverages (KLB) were characterized by low alcoholic grade (≤3.6%, v/v) and lactic and acetic acid concentrations. The beverages obtained from fermentation B had lower concentrations of sugars and higher microbial counts than the KLB obtained in fermentation A. Additionally, the KLB samples from fermentation B were the most aromatic and had the highest contents of alcohols, esters, aldehydes and organic acids, in contrast with the nonfermented juice and KLB from fermentation A. These results indicate the possibility of obtaining red table grape KLB with their own distinctive aromatic characteristics and high content in probiotic viable cells, contributing to the valorization of this fruit.

Two ethanol fermenting Saccharomyces cerevisiae were isolated from date juice and grapes and grown in YEPD medium. They were characterized for alcoholic fermentation using sugarcane molasses and their growth conditions were optimized with respect to pH and sugar concentration. Results revealed a temperature of 30ºC, pH 6.0 and 6.5% sugar concentration as optimum for fermentation. Stress tolerance tests showed that date juice isolate was highly tolerant to temperature, pH and high ethanol concentration in the medium. Under optimized conditions, S. cerevisiae isolated from date-juice produced 7.75% of ethanol in molasses as estimated by Conway method.Bangladesh J Microbiol, Volume 30, Number 1-2,June-Dec 2013, pp 49-54

Yeasts were isolated from four potential sources, sweet sorghum juice, sugar cane juice, grapes and rambutan. The 27 yeast isolates were tested for their ethanol tolerance (15% v/v of ethanol) and ethanol fermentation performance in a synthetic ethanol production medium (200 g/L of total sugar). Only five isolates, SCJ04KKU, SCJ07KKU, SCJ09KKU, SCJ14KKU and SSJ01KKU could tolerate 15% ethanol and produce ethanol at levels higher than 55 g/L. The ethanol production efficiency from sweet sorghum juice under high gravity (HG, 200 and 240 g/L of total sugar) and very high gravity (VHG, 280 g/L of total sugar) conditions of the five isolates was tested. Saccharomyces cerevisiae NP01 and S. cerevisiae ATCC4132 were used as reference strains. The results showed that the SSJ01KKU isolate gave the highest ethanol production efficiency under all conditions. Ethanol concentration (PE), yield (YP/S) and productivity (QP) values were 98.89 g/L, 0.50 and 1.18 g/L·h, respectively, with sugar consumption (SC) of 98.96% under the HG condition at 200 g/L of total sugar. Under the HG condition at 240 g/L of total sugar, the PE, YP/S and QP values were 118.12 g/L, 0.51 and 1.41 g/L·h, respectively, with the SC of 95.79%. These values were 82.29 g/L, 0.34 and 0.98 g/L·h, respectively, with the SC of 85.59% under the VHG condition. Addition of urea into the sweet sorghum juice under all conditions significantly shortened the fermentation time, resulting in increased QP values. Based on molecular taxonomic analysis of the five isolates using sequence analysis of the D1/D2 domain and the ITS1 and ITS2 regions, SSJ01KKU is S. cerevisiae, whereas SCJ04KKU, SCJ07KKU, SCJ09KKU and SCJ14KKU are Pichia caribbica.

Pineapples are a tropical fruit with high nutritional value and high vitamin and sugar contents. In this study, low-grade pineapples were fermented to produce vinegar using surface culture fermentation (SCF), which involved the addition of dragon fruit juice, to compare the quality and antioxidant activity of different preparations of vinegar. The highest acetic acid concentration (7.35%) was obtained from pineapple vinegar after 20 days of incubation. Vinegar made from mixed pineapple and dragon fruit juice without peel and vinegar with pineapple and dragon fruit juice with peel had acetic acid concentrations of up to 6.20% and 4.50%, respectively. The mixed-fruit vinegar of pineapple and dragon fruit juice with peel displayed the highest antioxidant activity at 210.74 µg/g TE, while no significant difference was found between the other two vinegars (189.52 vs. 187.91 µg/L TE). Notably, the volatile compounds detected in the vinegars were alcohols and esters, which may contribute to the distinct aroma. Overall, the addition of dragon fruit juice with peel to pineapple vinegar increased the phenolic content and antioxidant activity; however, fermentation was slightly slower than that of the other two test materials.

The effects of chamber drying under controlled temperature and moisture conditions and fermentation process on blueberry juices to obtain three wines were studied in this work. Drying was carried out with a view to increase the sugar content and obtain wines with an ethanol content similar to a commercial grape wine or to obtain sweet wines. Analyses included color parameters; browning index; and anthocyanin, flavonols, flavan-3-ol derivatives, and tannin concentrations, as well as vitamin C concentration and antioxidant activity. Based on the results, drying increases color and the concentration of anthocyanins and tannins most probably by the effect of dehydration of the berries and diffusion of the colored compounds from the skin to the pulp due to the structural alterations in their skin. In addition, drying decreases flavonols, flavan-3-ol derivatives, and vitamin C concentrations. The browning index, anthocyanins, and tannins decreased with the fermentation time, and vitamin C was constant with the fermentation time. The sensory analysis showed that the wines with the best sensory characteristics were those with residual sugar, partial fermented wines 1 and 2.

The conversion of grape juice to wine starts with complex yeast communities consisting of strains that have colonised the harvested grape and/or reside in the winery environment. As the conditions in the fermenting juice gradually become inhibitory for most species, they are rapidly overgrown by the more adaptable Saccharomyces strains, which then complete the fermentation. However, there are environmental factors that even Saccharomyces cannot cope with. We show that when the sugar content is extremely high, osmotolerant yeasts, usually considered as “spoilage yeasts“, ferment the must. The examination of the yeast biota of 22 botrytised Tokaj Essence wines of sugar concentrations ranging from 365 to 752 g∙L−1 identified the osmotolerant Zygosaccharomyces rouxii, Candida (Starmerella) lactis-condensi and Candida zemplinina (Starmerella bacillaris) as the dominating species. Ten additional species, mostly known as osmotolerant spoilage yeasts or biofilm-producing yeasts, were detected as minor components of the populations. The high phenotypical and molecular (karyotype, mtDNA restriction fragment length polymorphism (RFLP) and microsatellite-primed PCR (MSP-PCR)) diversity of the conspecific strains indicated that diverse clones of the species coexisted in the wines. Genetic segregation of certain clones and interactions (antagonism and crossfeeding) of the species also appeared to shape the fermenting yeast biota.

Chemical analysis of grape juice and wine has been performed for over 50 years in a targeted manner to determine a limited number of compounds using Gas Chromatography, Mass-Spectrometry (GC-MS) and High Pressure Liquid Chromatography (HPLC). Therefore, it only allowed the determination of metabolites that are present in high concentration, including major sugars, amino acids and some important carboxylic acids. Thus, the roles of many significant but less concentrated metabolites during wine making process are still not known. This is where metabolomics shows its enormous potential, mainly because of its capability in analyzing over 1000 metabolites in a single run due to the recent advancements of high resolution and sensitive analytical instruments. Metabolomics has predominantly been adopted by many wine scientists as a hypothesis-generating tool in an unbiased and non-targeted way to address various issues, including characterization of geographical origin (terroir) and wine yeast metabolic traits, determination of biomarkers for aroma compounds, and the monitoring of growth developments of grape vines and grapes. The aim of this review is to explore the published literature that made use of both targeted and untargeted metabolomics to study grapes and wines and also the fermentation process. In addition, insights are also provided into many other possible avenues where metabolomics shows tremendous potential as a question-driven approach in grape and wine research.

This study is the first report using tea fungus “kombucha” to ferment natural pomegranate juice to produce a fermented beverage with high content of glucuronic acid, as a human health beneficial component. We profited the acetic acid bacteria and yeasts symbiotic layer, which is well known in producing pharmaceutical beverages with considerable released organic acids such as glucuronic acid. Also, we used the natural pomegranate juice with high amount of carbohydrate and acid, as a favourable substrate for the fermentation process. The yield of glucuronic acid production was monitored by cultivating natural pomegranate juices under the 17 optimized-combinations of three distinct sucrose concentrations, fermentation temperatures, and processing time. The combinations were designated by applying the statistical response surface methodology method. The maximum amount of glucuronic acid 17.074g/l determined in the media with 8g/l supplementary sucrose after 14 days fermentation at 37°C, using high-performance liquid chromatography. Along with glucuronic acid production, effect of the three factors - sugar concentration, processing temperature and time - was also examined on changes of five physical and chemical properties of the fermented pomegranate juices, including; pH value, remained sucrose and reducing sugar content, kombucha layer biomass, and total acidity. Within 14-day fermentation process, the pH values showed decrease, the layers’ mass presented considerable increase, and the total acid content increased in the beverages. Overall, obtained data suggested that natural pomegranate juice can be a potential candidate for further development as a functional beverage to support the maximum human daily intake of glucuronic acid (45mg for a 70kg adult).

When we think of Saccharomyces cerevisiae, fermentation immediately comes to mind, but this is not the only trait that makes this yeast the organism of choice for bread, beer and wine production. The winemaking industry, for example, requires robust strains, capable of converting sugar to ethanol in challenging conditions; high osmotic stress and low pH in the initial grape must, followed by high ethanol concentration at the later stages of fermentation. Winemakers also look for ways of using fermentation to introduce aroma and flavour diversity to their wines as a means of improving style and for product differentiation. Choice of wine yeast from the plethora of strains available to winemakers is one way of achieving this, particularly with the new breed of interspecific hybrid yeast strains currently being generated.

<p>Samples from different industrial grape cultivars were collected during the vintage season from the vineyard of the winery (the «Shabo» winery Company, located in the Odesa region, Ukraine). The following industrial cultivars of grapes were selected for the research: Chardonnay, Cabernet Sauvignon, Merlot, Sauvignon, Riesling Rhenish, Aligote, Rkatsiteli, Bastardo, Traminer, Telti Kuruk, Grinosh.</p> <p>The grape cultivars were cultivated on the sandy soils in the district located between the Black Sea and the Dnestrovsky estuary. Grape must derived from different grape cultivars was placed into sterile glass flasks to half of the 450ml flask volume. Each flask was carefully closed with a rubber stopper with an injection needle in it. During the fermentation process, it was necessary to remove carbon dioxide, which was present as a result of active anaerobic fermentation processes in the grape must. At the end of grape must fermentation, pure yeast cultures were isolated using traditional microbiological methods by consistent inoculation of a sample into a Petri dish with a few modifications of nutrient selective agar for yeast isolation and cultivation. Primary yeast isolation was carried out using Inhibitory Mold Agar medium (Becton Dickinson Company, USA).</p> <p>The yeast culture morphological properties were analyzed after the primary yeast culture isolation. Yeasts were identified by polymerase chain reaction (PCR) using universal yeast primers. After yeast culture identification, the next step in yeast cultivation was carried out on Wort Agar medium (Becton Dickinson Company, USA). Each isolated, and identified yeast culture was deposited in the Genebank of Japan, MAFF culture Collection, Tsukuba, Ibaraki, Japan and (NCYC) - Yeast Culture Collection (National Collection of Yeast Cultures, Institute of Food Research, Norwich, United Kingdom). Each yeast culture was tested for technological characteristics such as growth resistance to high temperature (+42°C) and low temperature (+6°C), growth at low pH 2.6–3.0 (acid resistance), growth in the presence of 5, 10, and 15% ethanol (ethanol resistance), and growth in the presence of high concentration potassium bisulfite (bisulfite resistance). Hydrosulfide synthesis (H₂S gassing production) was studied in addition.</p> <p>Parameters of cellular metabolism in yeast suspension, such as concentration of nitrogen, protein, triglicerides, enzymatic activity and total sugar (which include glucose, fructose, and galactose) were determined. Macro- and micro-element concentrations in fermented grape must, which contained pure yeast culture was determined and included: potassium, sodium, calcium, phosphorus, magnesium, iron, chlorides. In addition to identifying parameters of macro- and micro- element concentration in grape must during and following fermentation based on a principle of photometric analysis, carried out using a biochemical analyser Respons-920 (DiaSys Diagnostic Systems GmbH, Germany).</p> <p>Laboratory selected <em>Saccharomyces cerevisiae </em>wine yeast showed high enzymatic activity with short lag phase. Since of fermentation started on third day concentration of Triglicerides, Protein (total), Potassium and Sodium increased and then level of Protein (total) on the 5th day of fermentation twice decreased. Trigliceride concentration on the 5th day of fermentation continued to increase. Concentration of Iron on the 5th day of fermentation increase in geometrical progression, concentration increase in 4-5 times. Contrary Chloride concentration on the 5th day of fermentation decreased in 3-4 times. Enzymatic activity on 3rd day of fermentation maximal for Lactate Dehydrogenase, Alanine aminotransferase, Aspartate aminotransferase, Phosphatase. Since of 5th day of fermentation Enzymatic activity for Lactate Dehydrogenase, Alanine aminotransferase, Aspartate aminotransferase 3-4 times. Especially level of Phosphatase activity very decreased in 6-7 times. Comparative assessment between our Laboratory selected <em>Saccharomyces cerevisiae</em> yeast culture and Dry active commercial <em>Saccharomyces cerevisiae</em> yeast culture did not showed any difference in enzymatic activity. Both groups showed high enzymatic activity on the third day from the start of fermentation and decreasing on the fifth day since of fermentation started.</p> <p><em> Key words: wine yeast, enzymatic activity, cellular metabolism</em><em>, </em><em>Saccharomyces cerevisiae</em><em>.</em><em></em></p>

Yeasts are able to act as biosorbents, as their cell wall includes several components capable of binding organic xenobiotic compounds that can potentially be removed during various fermentation processes. In the present investigation, two novel Saccharomyces cerevisiae strains (LMBF-Y 16 and LMBF-Y-18), previously isolated from grapes, were studied regarding their physiological behavior (dry cell weight—DCW production, substrate uptake, and ethanol and glycerol biosynthesis) during fermentations of grape must, in some cases enriched with commercial glucose and fructose (initial total sugar concentration approximately 150 and 250 g/L, respectively). Myclobutanil (a chiral triazole fungicide broadly used as a protective agent of vine) was also added to the culture media at various concentrations in order to assess the ability of the yeasts to simultaneously perform alcoholic fermentations and detoxify the medium (i.e., to remove the fungicide). In the first set of experiments and for both tested strains, trials were carried out in either 250 mL or 2.0 L agitated shake flasks in either synthetic glucose-based experiments or grape musts. Since the results obtained in the trials where the cultures were placed in 2.0 L flasks with grape musts as substrates were superior in terms of both DCW and ethanol production, these experimental conditions were selected for the subsequent studies. Both strains showed high fermentative efficiency, producing high amounts of DCW (9.5–10.5 g/L) in parallel with high ethanol production, which in some cases achieved values very close to the maximum theoretical ethanol production yield (≈0.49 g of ethanol per g of sugar). When using grape must with initial total sugars at approximately 250 g/L (very high gravity fermentation media, close to winemaking conditions), significantly high ethanol quantities (i.e., ranging between 105 and 123 g/L) were produced. Myclobutanil addition slightly negatively affected sugar conversion into ethanol; however, in all cases, ethanol production was very satisfactory. A non-negligible myclobutanil removal during fermentation, which ranged between 5%–27%, as a result of the adsorptive or degradative capacity of the yeast was also reported. The presence of myclobutanil had no effect on DCW production and resulted in no significant differences in the biosynthesis of glycerol. Therefore, these newly isolated yeast strains could be excellent candidates for simultaneous high ethanol production and parallel pesticide removal in a general biorefinery concept demonstrating many environmental benefits.

On the frame of this research survey, a novel potentially probiotic strain (Lactobacillus paracasei SP5) recently isolated from kefir grains was evaluated for chokeberry juice fermentation. Chokeberry juice was retrieved from the variety Aronia melanocarpa, a plant known to provide small, dark berries and to be one of the richest sources of antioxidants. The juice was subsequently fermented inoculating L. paracasei SP5 for 48 h at 30 °C. The fermented juices were left at 4 °C and tested regarding microbiological and physicochemical characteristics for 4 weeks. The potentially probiotic strain was proved capable of performing lactic acid fermentation at 30 °C. Cell viability of L. paracasei was detected in high levels during fermentation and the whole storage period, while the fermented juice showed higher levels of viability in juice with 40.3 g/L of initial sugar concentration. No ethanol was detected in the final fermented juice. Fermented chokeberry juice was characterized by aromatic desirable volatiles, which were retained in adequate levels for the whole storage period. Specifically, the occurrence of organic esters detected in fermented juices is considered as positive evidence of the provision of fruity and floral notes to the final product. During storage, total phenolics content and antioxidant activity were observed in higher levels in fermented chokeberry juice compared with non-fermented juice. Subsequently, fermentation of chokeberry juice by potentially probiotic lactic acid bacteria could provide high industrialization potential, providing the market with a nutritional beverage of good volatile quality with an enhanced shelf-life compared with an unfermented fresh juice.

ABSTRACTOptimizing ethanol yield during fermentation is important for efficient production of fuel alcohol, as well as wine and other alcoholic beverages. However, increasing ethanol concentrations during fermentation can create problems that result in arrested or sluggish sugar-to-ethanol conversion. The fundamental cellular basis for these problem fermentations, however, is not well understood. Small-scale fermentations were performed in a synthetic grape must using 22 industrialSaccharomyces cerevisiaestrains (primarily wine strains) with various degrees of ethanol tolerance to assess the correlation between lipid composition and fermentation kinetic parameters. Lipids were extracted at several fermentation time points representing different growth phases of the yeast to quantitatively analyze phospholipids and ergosterol utilizing atmospheric pressure ionization-mass spectrometry methods. Lipid profiling of individual fermentations indicated that yeast lipid class profiles do not shift dramatically in composition over the course of fermentation. Multivariate statistical analysis of the data was performed using partial least-squares linear regression modeling to correlate lipid composition data with fermentation kinetic data. The results indicate a strong correlation (R2= 0.91) between the overall lipid composition and the final ethanol concentration (wt/wt), an indicator of strain ethanol tolerance. One potential component of ethanol tolerance, the maximum yeast cell concentration, was also found to be a strong function of lipid composition (R2= 0.97). Specifically, strains unable to complete fermentation were associated with high phosphatidylinositol levels early in fermentation. Yeast strains that achieved the highest cell densities and ethanol concentrations were positively correlated with phosphatidylcholine species similar to those known to decrease the perturbing effects of ethanol in model membrane systems.

ABSTRACTOver recent decades, the average ethanol concentration of wine has increased, largely due to consumer preference for wine styles associated with increased grape maturity; sugar content increases with grape maturity, and this translates into increased alcohol content in wine. However, high ethanol content impacts wine sensory properties, reducing the perceived complexity of flavors and aromas. In addition, for health and economic reasons, the wine sector is actively seeking technologies to facilitate the production of wines with lower ethanol content. Nonconventional yeast species, in particular, non-Saccharomycesyeasts, have shown potential for producing wines with lower alcohol content. These yeast species, which are largely associated with grapes preharvest, are present in the early stages of fermentation but, in general, are not capable of completing alcoholic fermentation. We have evaluated 50 different non-Saccharomycesisolates belonging to 24 different genera for their capacity to produce wine with a lower ethanol concentration when used in sequential inoculation regimes with aSaccharomyces cerevisiaewine strain. A sequential inoculation ofMetschnikowia pulcherrimaAWRI1149 followed by anS. cerevisiaewine strain was best able to produce wine with an ethanol concentration lower than that achieved with the single-inoculum, wine yeast control. Sequential fermentations utilizing AWRI1149 produced wines with 0.9% (vol/vol) and 1.6% (vol/vol) (corresponding to 7.1 g/liter and 12.6 g/liter, respectively) lower ethanol concentrations in Chardonnay and Shiraz wines, respectively. In Chardonnay wine, the total concentration of esters and higher alcohols was higher for wines generated from sequential inoculations, whereas the total concentration of volatile acids was significantly lower. In sequentially inoculated Shiraz wines, the total concentration of higher alcohols was higher and the total concentration of volatile acids was reduced compared with those in controlS. cerevisiaewines, whereas the total concentrations of esters were not significantly different.

In this experiment, the effect of pulsed electric field (PEF) technology on the extractability of anthocyanins and polyphenols in early-harvested Sangiovese red grapes (16.9°Bx sugar, 3.26 pH, and 10.4 g/L titratable acidity) from Emilia Romagna (Italy) was investigated. Electric field strengths were in the range of 0.9–3 kV/cm, generated by the application of short, high-voltage pulses, and the grapes were subjected to specific energies from 10.4 to 32.5 kJ/kg immediately after crushing and destemming to produce a pre-fermentative pulsed electric field treatment on a pilot scale. Grape musts and wines were analyzed for color components and polyphenols content from pressing of juices up to 3 months from the end of the fermentation of wines. Furthermore, the freshly-fermented wines were subjected to accelerated aging conditions (i.e., warming under 40 °C for 32 days) to simulate the evolution of color parameters with time. The color intensity was generally higher in treated musts and wines compared to the control, further increased by raising the intensity of the electric field. Results suggested the potentialities of pulsed electric fields (PEFs) as a mild pre-fermentative process to assist maceration and to increase the polyphenolic content of musts obtained by early-harvested Sangiovese grapes.

Organic acids represent naturally occurring compounds that are found in many types of food and beverages, with important functions in defining products’ final quality. Their proportions in wine are dependent on grape composition and winemaking conditions (temperature, pH levels, oxygen, and carbon dioxide concentration). Therefore, this article studied the influence of different fermentation conditions (200 hL tanks vs. 50 L glass demijohns) and various yeasts on the evolution of the main organic acids during alcoholic fermentation of “Aligoté” wines. The fermentation lasted 22 days and samples were collected daily. Laboratory analyses were quantified according to the International Organization of Vine and Wine recommendations. High-performance liquid chromatography for the identification and quantification of organic acids was used. The data showed the important effect of winemaking conditions on sugar consumption, density or acidity values, and sensory characteristics. Significant differences in organic acid concentrations (especially for tartaric acid) were obtained for all variants, depending on the fermentation conditions, inoculated yeast and the sampling moment. The quantities of most of the identified organic acids were generally significantly increased when glass vessels were used, compared to those fermented in tanks. Most organic acids concentrations were favored by lower pH and showed higher values at lower temperatures.

The effects of climate change are posing major challenges for winemaking, especially in warm regions. Rising temperatures are leading to a considerable increase in sugar content and a reduction in the acidity of grape juices. This has prompted the search for new chemical and biological tools to reduce these effects in winemaking. Lachancea thermotolerans represent an interesting tool for wine improvement, due to their effects on lactic acid, ethanol, and volatile acidity. In this study, we evaluated the sequential inoculation of Lachancea thermotolerans (AEB Italia) + Saccharomyces cerevisiae as an alternative to the addition of water and tartaric acid in the vinification of over-ripened wine must (Tempranillo cv.). To do so, we conducted a micro vinification experiment that included three experimental treatments (replicated four times); AAS = Addition of water + tartaric acid fermented with Saccharomyces cerevisiae, ALS = sequential fermentation with Lachancea thermotolerans followed by Saccharomyces cerevisiae and SAC = Only inoculation with Saccharomyces cerevisiae (control). We measured the lactic acid, malic acid, acetic acid, total acidity, pH, glucose + fructose, and ethanol concentration of the resulting wines. We found a higher lactic acid content and total acidity in ALS wines (2.2 g/L y 6.65 g/L) compared with AAS (0.13 g/L y 5.75 g/L) and SAC wines (0.18 g/L y 5.37 g/L). ALS wines showed a significantly lower pH (4.00) than SAC wines (4.19) but did not differ significantly from AAS wines (3.19). Finally, acetic acid was higher in ALS wines (0.89 g/L) than AAS wines (0.64 g/L) but did not differ significantly from SAC wines (1.02 g/L). Our results point out that the use of Lachancea thermotolerans in a sequential inoculation with Saccharomyces cerevisiae is an alternative to the addition of water and tartaric acid in the vinification of over-ripened Tempranillo musts.

The present study describes the application of an acid tolerant and potentially probiotic L. paracasei SP3 strain, recently isolated from kefir grains, in the production of a novel functional beverage based on the fermentation of pomegranate juice. The fermentation ability of the novel strain was assessed during pomegranate juice fermentations at 30 °C for 24 h and storage at 4 °C for 4 weeks. Various parameters were assessed such as residual sugar, organic acid and alcohol levels, total phenolics content, antioxidant activity, astringency, cell viability, and consumer acceptance. Residual sugar was decreased by approximately 25%, while respectable amounts of lactic acid were determined (4.8 g/L) on the 28th day of storage, proving that the novel strain was effective at lactic acid fermentation. The concentration of ethanol was maintained at low levels (0.3–0.4 % v/v) and low levels of acetic acid were detected (0.6 g/L). The viability of L. paracasei SP3 cells retained high levels (>7 log cfu/mL), even by the 4th week. The total phenolic content (123.7–201.1 mg GAE/100 mL) and antioxidant activity (124.5–148.5 mgTE/100 mL) of fermented pomegranate juice were recorded at higher levels for all of the studied time periods compared to the non-fermented juice. The employment of the novel strain led to a significant reduction in the levels of hydrolysable tannins (42%) in the juice, reducing its astringency. The latter was further proven through sensorial tests, which reflected the amelioration of the sensorial features of the final product. It should be underlined that fruit juices as well as pomegranate juice comprised a very harsh food matrix for microorganisms to survive and ferment. Likewise, the L. paracasei SP3 strain showed a significant potential, because it was applied as a free culture, without the application of microencapsulation methods that are usually employed in these fermentations, leading to a product with possible functional properties and a high nutritive value.

In this study ‘Debina’ (Zitsa, Ioannina, Greece) wines (WD: white dry and WDS: white dry sparkling) were examined in terms of their quality (chemical, physicochemical and sensory) parameters. The concentration of alcohol in both wines was practically stable (p>0.05) from the stage of fermentation to packaging and specifically, the rates were 12 and 11.4% vol for WD and WDS wines, respectively. Total sugar concentrations were equal to 1.4 g/l and 1.2 g/l in WD and WDS wines, respectively. WD wine had final total acidity, volatile acidity and pH values of 4.9 g/l tartaric acid, 0.19 g/l acetic acid and 3.15, respectively. While, WDS wine had final total acidity, volatile acidity and pH value of 6.0 g/l tartaric acid, 0.15 g/l acetic acid and 3.11, respectively. Τotal sulfite content of wine samples decreased and final values of 90.6 mg/l and 99.8 mg/l, in the last production stage, for the WD and WDS wines, were recorded, respectively. According to the data of the study, all wines received a high acceptability sensory score, as judged by the panellists. It must be reported that the sensory scores were on average, 1 point higher for the WDS wines, as compared to the WD wines.

В статье представлены результаты исследований по влиянию различных сахаросодержащих компонентов, используемых для приготовления игристых вин, на их качество, в том числе типичные свойства. Показано, что практически все исследуемые опытные игристые вина, приготовленные с использованием различных сахаросодержащих компонентов виноградного происхождения, имели более высокую дегустационную оценку по сравнению с контролем (приготовленным с использованием тиражного ликёра). Установлено, что игристые вина, выработанные на основе недобродов, имели более высокие показатели пенистых свойств, лучшую насыщенность диоксидом углерода, высокое содержание общего и связанного диоксида углерода, более высокую массовую концентрацию фенольных веществ и интенсивность окраски. Красные игристые вина, приготовленные на основе недобродов, содержали меньшее количество альдегидов, по сравнению с контрольными образцами, приготовленными с использованием тиражного ликёра. Образцы игристых вин, приготовленные с использованием сусла виноградного концентрированного, имели более высокую массовую концентрацию титруемых кислот, что обусловлено концентрированием в процессе вакуумирования сусла не только сахаров, но и органических кислот и ряда других веществ экстракта. Использование ликёрного виноматериала при приготовлении красных игристых вин практически не изменяло массовую концентрацию фенольных веществ, но внесло новую гамму в букет и вкус игристого вина. Следует отметить, что каждый из исследуемых сахаросодержащих компонентов виноградного происхождения имеет свои преимущества и недостатки. И в зависимости от поставленных задач может применяться для приготовления высококачественных игристых вин.The effects of various sugar-containing components used in production of sparkings on their quality were studied. Practically all study sparkings manufactured with the use of various sugar-containing components of grape origin had higher tasting scores compared to controls where tirage liqueur was used. Sparklings manufactured from materials in which fermentation was not allowed to complete were superior in foaming properties and saturation with carbon dioxide, in addition to high levels of total and bound carbon dioxide, increased levels or phenolic substances, and a more intense color. Red sparkings from materials with incomplete fermentation had lower aldehyde levels in comparison to controls. Sparklings manufactured with the use of vacuum must were higher in titratable acidity since, besides sugars, vacuumization involves concentration of organic acids and other extract components. The use of liqueur wine material for manufacturing of red sparklings practically did not change the levels of phenolic components but added new aromas and flavours. Each of the study sugar-containing components of grape origin has advantages and disadvantages and can be used to manufacture quality sparklings with different tasks in mind.

Local vegetable raw materials have good prospects in developing new balanced foods with high nutrition value. As a rule, local vegetable raw materials are environmentally friendly and contain a wide range of biologically and physiologically active agents. Fruit liqueurs possess a balanced chemical composition and a high nutrition and physiological value. They produce a recreational or preventive effect on human organism. However, the level of consumption of fresh fruit and berries remains low because of their seasonality. Moreover, the range of fruit and berry products is usually very poor, and very few of them are beneficial for human health and can prevent diseases. The research featured the formulations and production technology for new multicomponent fruit liqueurs of high nutrition value. The liqueurs were made from Dagestan garden and wild fruit and berries with a high content of macro- and micronutrients. The nutrition value and quality of the liqueurs were estimated according to the mass concentration of sugars, titrable acids, vitamins C and P, β-carotene, mineral elements, and phenolic and pectinaceous substances. The sensory properties were measured according to a 10-score system. The paper introduces formulations and technologies for three types of liqueur (Zhivitel’naya, Garmoniya, and Uslada). They involved alcoholized and fermentation-alcoholized juices of grapes, fruit and berry fruit drinks and extracts that mutually complemented each other according to valuable components. The new liqueurs have a high nutrition since 100 cm3 of the liqueur per day partially satisfies the daily requirements for some nutrients: phenolic substances – for 37.5–55.0%; pectinaceous substances – 17.5–32.5%; in vitamin C – 13.7–38.7%, β-carotene – 17.6–43.0%, and a microelement iron – for 14.0–26.0%. The multicomponent fruit liqueurs possess a wide range of biologically and physiologically active agents. As a result, they improve physical activity and increase resistance to bad ecological environment.

The maguey mezcalero potosino (Agave salmiana) is a natural resource of great regional importance, used for the manufacture of mezcal and other alcoholic beverages by fermentation of simple sugars and sugars released by fructans hydrolysis. For this, only the stem and base of leaves is used, which results on wasting the other sections that represent up to 40% of weight of the complete plant. The objective of this study was to identify and quantify the fermentable carbohydrates present in raw and hydrolysed juices of the stem and four sections of the whole leaves (base, neck, wings and apex) of maguey plants with representative specimens of three stages of maturity with industrial importance. For carbohydrate characterization, high resolution liquid chromatography (HLPC) was used. The sugars found as fructose, glucose, sucrose, xylose and maltose coincide with those identified in other Agave species, significant differences are observed depending on the section and the state of maturity of the plant, the sugar concentration in the discarded sections of the plant (fructose: 5.79 – 4.80 mg mL-1) are considerable and statistically similar to those found in the stem (fructose: 2.49 mg mL-1) and the base of the leaf (fructose: 3.16 mg mL-1) (structure used in the mezcal production process). The results obtained can promote the integral use of the plant and diversification towards other industries such as pharmaceuticals, biofuels or foods.

<p style="text-align: justify;">In this work, encapsulated whole cells of <em>S. cerevisiae</em> in calcium-alginate gel (<em>S. cerevisiae</em>-CAG) were used for the treatment of sluggish and stuck fermentation in vinification. <em>S. cerevisiae</em>-CAG can be applied into the must grape and then they can be withdrawn easily at the end of alcoholic fermentation. For the treatment of sluggish and stuck fermentations, <em>S. cerevisiae</em>-CAG were applied following two steps: firstly <em>S. cerevisiae</em>-CAG were introduced into permeable bags and they were activated in a liquid medium; secondly, after an activation period of eight hours, the immobilized yeast cells were introduced into the fermentation tank. In this work, the obtained results for the treatment of sluggish and stuck fermentations in several French and Portuguese wineries are presented. Preliminary results obtained in micro-vinification conditions have shown that the use of immobilized yeast was better than traditional method (which uses free cells) for the treatment of stuck and sluggish fermentations. The large success rate of immobilized yeast can be explained by one adaptation step of yeast cells to high ethanol concentrations during the immobilization process. The application of immobilized cells of <em>S. cerevisiae</em> under real conditions of vinification has shown a consumption rate of 2.8 g/L per day of reducing sugar with a concentration of 5 million of viable cells per mL. It was never observed any increase of the volatile acidity or of other undesirable compounds.</p>

In modern wine-making technology, there is an increasing concern in relation to the preservation of the biodiversity, and the employment of “new”, “novel” and wild-type Saccharomyces cerevisiae strains as cell factories amenable for the production of wines that are not “homogenous”, expressing their terroir and presenting interesting and “local” sensory characteristics. Under this approach, in the current study, several wild-type Saccharomyces cerevisiae yeast strains (LMBF Y-10, Y-25, Y-35 and Y-54), priorly isolated from wine and grape origin, selected from the private culture collection of the Agricultural University of Athens, were tested regarding their biochemical behavior on glucose-based (initial concentrations ca 100 and 200 g/L) shake-flask experiments. The wild yeast strains were compared with commercial yeast strains (viz. Symphony, Cross X and Passion Fruit) in the same conditions. All selected strains rapidly assimilated glucose from the medium converting it into ethanol in good rates, despite the imposed aerobic conditions. Concerning the wild strains, the best results were achieved for the strain LMBF Y-54 in which maximum ethanol production (EtOHmax) up to 68 g/L, with simultaneous ethanol yield on sugar consumed = 0.38 g/g were recorded. Other wild strains tested (LMBF Y-10, Y-25 and Y-35) achieved lower ethanol production (up to ≈47 g/L). Regarding the commercial strains, the highest ethanol concentration was achieved by S. cerevisiae Passion Fruit (EtOHmax = 91.1 g/L, yield = 0.45 g/g). Subsequently, the “novel” strain that presented the best technological characteristics regards its sugar consumption and alcohol production properties (viz. LMBF Y-54) and the commercial strain that equally presented the best previously mentioned technological characteristics (viz. Passion Fruit) were further selected for the wine-making process. The selected must originated from red and white grapes (Assyrtiko and Mavrotragano, Santorini Island; Greece) and fermentation was performed under wine-making conditions showing high yields for both strains (EtOHmax = 98–106 g/L, ethanol yield = 0.47–0.50 g/g), demonstrating the production efficiency under microaerophilic/anaerobic conditions. Molecular identification by rep-PCR carried out throughout fermentations verified that each inoculated yeast was the one that dominated during the whole bioprocess. The aromatic compounds of the produced wines were qualitatively analyzed at the end of the processes. The results highlight the optimum technological characteristics of the selected “new” wild strain (S. cerevisiae LMBF Y-54), verifying its suitability for wine production while posing great potential for future industrial applications.

Изменение климата приводит к повышению содержания сахара в виноградном сусле, снижению концентрации органических кислот, прекурсоров ароматобразующих веществ и т.д. и, как следствие, к повышение содержания алкоголя, нарушению баланса вкуса и искажению аромата вина и другие проблемы. Технологические подходы, предлагаемые для их решения, хотя и позволяют достичь цели, но часто негативно влияют на качество вина. Перспективной альтернативой является использование микроорганизмов, не относящихся к Saccharomyces , продукты метаболизма которых - глицерин, кислоты, маннопротеины, полисахариды и др. - оказывают влияние на органолептические характеристики вина. Так, использование дрожжей Candida spp., Metschnikowia spp., Lachancea spp. способствует снижению содержания этанола в винах на 1,5-2,0% об. Hansensiaspora spp., Pichia spp., Starmerella spp., Torulaspora spp. и др. отличаются высокой способностью к синтезу глицерина и полисахаридов. Использования консорциума дрожжей Saccharomyces и не- Saccharomyces для брожения сусла приводит к усилению ароматических и вкусовых характеристик вин. Отмечено, что среди не сахаромицетов присутствуют организмы, синтезирующие малые количества уксусной кислоты и ацетальдегида, что благоприятно влияет на качество получаемых вин. Настоящая работа является результатом систематизации информации, касающейся некоторых аспектов использования дрожжей несахаромицетов в винодельческой промышленности, их влияния на химический состав вин. Climate change leads to an increase in the sugar content of grape must, a decrease in the concentration of organic acids, precursors of aroma-producing substances, etc. and, as a consequence, to an increase in the alcohol content, flavor imbalance and distortion of wine aroma and other problems. Even though technological approaches proposed for solution allow to achieve the goal, they often negatively affect the quality of wine. A promising alternative is using of non- Saccharomyces microorganisms with their metabolic products - glycerin, acids, mannoproteins, polysaccharides, etc. affecting the organoleptic characteristics of wine. So, using of yeast Candida spp., Metschnikowia spp., Lachancea spp. helps to reduce ethanol content in wines by 1.5-2.0% by volume. Hansensiaspora spp., Pichia spp., Starmerella spp., Torulaspora spp. and others are distinguished by a high ability to synthesize glycerin and polysaccharides. Using of yeast consortium of Saccharomyces and non- Saccharomyces for must fermentation leads to an increase in the aroma and flavor characteristics of wines. It was noted that among the non-saccharomycetes there are organisms that synthesize small amounts of acetic acid and acetaldehyde, favorably affecting the quality of wines obtained. This work is the result of information systematization concerning some aspects of using non-Saccharomyces in winemaking industry, their effect on the chemical composition of wines.

Aim: A Saccharomyces uvarum isolate was assessed for its ability to metabolize acetic acid present in juice and during the fermentation of partially dehydrated grapes. The impact on other yeast metabolites was also compared using an S. uvarum isolate and an S. cerevisiae wine yeast. The upper limit of fruit concentration that allowed the S. uvarum isolate to ferment wines to < 5 g/L residual sugar was defined.Methods and results: Cabernet franc grapes were partially dehydrated to three different post-harvest sugar targets (24.5 °Brix, 26.0 °Brix, and 27.5 °Brix) along with non-dehydrated grapes (21.5 °Brix control). Musts from all treatments were vinified with either the S. uvarum isolate CN1, formerly identified as S. bayanus, or S. cerevisiae EC1118. All wines were successfully vinified to less than 5 g/L residual sugar. Fermentation kinetics between the two yeasts were similar for all wines other than 27.5 °Brix, where CN1 took three days longer. During fermentation with CN1, acetic acid peaked on day two, then decreased in concentration, resulting in final wine acetic acid lower than that measured on day two. Wines fermented with EC1118 showed an increase in acetic acid over the time-course of fermentation. Significantly lower wine oxidative compounds (acetic acid, acetaldehyde and ethyl acetate) and higher glycerol resulted in wine produced with CN1 in comparison to EC1118. Both yeasts produced comparable ethanol at each Brix level tested. Further studies showed that CN1 lowered acetic acid seven-fold from 0.48 g/L in juice to 0.07 g/L in wine whereas EC1118 reduced acetic acid to 0.18 g/L.Conclusions: The autochthonous S. uvarum yeast isolate successfully fermented partially dehydrated grapes to < 5 g/L sugar up to 27.5 ºBrix. The consumption rate of acetic acid was faster than its production during fermentation, resulting in low acetic acid, acetaldehyde and ethyl acetate in wine in comparison to a commercial S. cerevisiae yeast while consistently producing higher glycerol.Significance and impact of the study: The S. uvarum yeast isolate can metabolize acetic acid during fermentation to significantly lower acetic acid, ethyl acetate and acetaldehyde in wine. It can also reduce acetic acid by seven-fold from the starting juice to the finished wine, which could have potential application for managing sour rot arising in the vineyard or during the dehydration process in making appassimento-style wines.

Cultivo de gránulo de kéfir en zumo de uvas tintas Kefir culture pellet red grape juice Santos Pedraza Guevara Universidad de Vigo – España Universidad Nacional Toribio Rodríguez de Mendoza de Amazonas – Perú DOI: https://doi.org/10.33017/RevECIPeru2015.0006/ Resumen Este estudio forma parte de investigaciones que desarrollan en el área de Bromatología, Dpto. de Química Analítica y Alimentaria, Universidad de Vigo Campus Ourense, España; cuyo objetivo final es producir bebidas fermentadas con propiedades probióticas con gránulos de kéfir en zumo de uvas tintas y blancas con ello se estaría valorizando uvas no vinificables con escaso valor comercial o consumo directo. Y obteniendo bebidas que combine efectos saludables de uva tinta fundamentalmente polifenoles con carácter antioxidante y beneficios funcionales del kéfir. Se estudió las cinéticas de principales variables del proceso fermentativo, se analizó el efecto del cambio de sustrato sobre la fermentación y poblaciones microbianas del kéfir. El procedimiento se llevó a cabo de la siguiente manera: se activaron el gránulo, se llevó a cabo una serie de 4 cultivos sucesivos. En cada uno se incubó el gránulo en zumo de uva tinta no estéril durante 24 h. Se separó el gránulo y se sustituyó zumo fermentado por zumo fresco. A las 12 horas de incubación y en el zumo fermentado tras 24 h se tomaron muestras, se centrifugaron para separar biomasa a partir de la cual se determinó la concentración de células libres y las UFC de cada uno de los 3 grandes grupos microbianos bacterias ácido lácticas, acéticas y levaduras, y en los sobrenadantes se analizaron pH, azúcares y metabolitos mediante HPLC. Adicionalmente se tomaron muestras al inicio y final de cada fermentación para determinar conteos BLA, BAC y LEV. De acuerdo a los resultados obtenidos, parece lógico utilizar el producto fermentado obtenido para una doble finalidad. La bebida obtenida por fermentación de zumo de uvas tintas con gránulos de kéfir, podría ser utilizada como una nueva bebida alcohólica por su elevado contenido en etanol (~33 g/L), previa separación de las células. Las células probióticas obtenidas, con alta viabilidad podrían ser utilizadas para producir un suplemento alimenticio, por los potenciales beneficios, que su uso podría producir sobre la salud de los consumidores del producto. Las concentraciones de biomasa libre obtenidas en el zumo durante los dos primeros pases, oscilan entre 1 y 2 g/L, pero en el tercer pase, se obtiene un valor máximo en torno a los 6 g/L. Este repunte en la producción de biomasa libre en el tercer pase, se corresponde también con el incremento del consumo de azúcares (glucosa y fructosa) a partir del tercer pase (subcultivo), posiblemente porque en esta etapa ya los gránulos de kéfir estaban adaptados a la composición del nuevo medio de fermentación con respecto al medio de activación. En cuanto a la formación de productos, llama poderosamente la atención, en comparación con los cultivos en suero de leche, las elevadas producciones de etanol que superan en 4 veces, las obtenidas en suero de leche y el consumo completo del ácido láctico del medio de fermentación, la total ausencia de ácido acético, además de la producción de cantidades apreciables de glicerol desde 2 g/L en el primer pase hasta aproximadamente 6 g/L, en el cuarto pase. Todo ello sugiere una mayor actividad de la población levaduriforme con respecto a las poblaciones de bacterias lácticas y acéticas. El nivel de pH inicial del zumo de uva tinta (~3,8), adecuado para el desarrollo de levaduras e inadecuado para el desarrollo de bacterias lácticas (BAL) y acéticas (BAC) apoyan esta hipótesis. La desaparición de ácido láctico del medio de fermentación sugiere más su utilización por parte de las levaduras asimiladoras de este ácido, que a un reconsumo por parte de las BAL debido a las altas concentraciones iniciales de azúcares (glucosa y fructosa) en el medio de fermentación. Descriptores: bebida probiótica, fermentación, kéfir, uvas tintas. Abstract This study is part of research developed in the area of ​​Food Science, Department of Analytical Chemistry and Food, University of Vigo Ourense Campus, Spain.; whose ultimate goal is to occurring fermented beverages with probiotic properties with kefir grains juice of red and white grapes would thus not valuing vinificables grapes with low commercial value or direct consumption. And obtaining healthy drinks combining ink effects mainly polyphenol grape character with antioxidant kefir and functional benefits. Kinetics main fermentation process cash studied, the effect of change on the fermentation substrate and microbial populations analyzed kefir. The procedure was carried out as follows: the pellet was activated, conducted a sequence of four successive crops. In each one the juice grain was incubated of nonsterile red grape during 24 h. The grain separated and juice fermented by fresh juice was replaced. At 12 hours of incubation and in the juice fermented after 24 h samples were taken, centrifuged to separate biomass from which the concentration of free cells and CFU of each of the 3 great microbial groups lactic acid bacterium was determined, acetic acid and yeast, and the supernatants were analyzed pH, sugars and metabolites by HPLC. Additionally samples at the beginning and end of each fermentation to determine counts BLA, BAC and LEV were taken. According to the obtained results, it seems logical to use the product fermented obtained for one double purpose. The beverage fermented juice kefir grains inks grapes, could be used as a new alcoholic beverage due to its high ethanol content (~ 33 g/L), after separation of the cells. The probiotic cells obtained with high viability could be used to occurring a food supplement, the potential benefits that their use might have on the health of consumers of the product. Biomass concentrations of free juice obtained during the first two passes, between 1 and 2 g/L, but in the third pass, a maximum value at about 6 g/L is obtained. This rise in the production of free biomass in the third pass, also corresponds with the increased consumption of sugars (glucose and fructose) from the third passage (subculture), possibly because at this stage and kefir grains were adapted to the composition of the fermentation medium again from the average activation. As for the formation of products, catches the eye, compared to whey crops, high yields of ethanol exceeding 4 times, the whey obtained full consumption and lactic acid medium fermentation, the total absence of acetic acid in addition to the production of significant amounts of glycerol from 2 g/L in the first pass to about 6 g/L, in the fourth pass. This suggests an increased activity of the population regarding yeast populations of lactic and acetic bacterium. The initial pH level of red grape juice (~ 3.8), suitable for the growth of yeasts and unsuitable for the development of lactic acid bacterium (LAB) and acetic acid (BAC) support this hypothesis. The disappearance of lactic acid from the fermentation medium suggests further use by the assimilating yeasts of this acid, a reconsumo by LAB due to high initial concentrations of sugars (glucose and fructose) in the fermentation. Keywords: probiotic drink, fermentation, kefir, red grapes.

Wine is a worldwide known beverage, and even though its consumption has been associated with the reduction of heart diseases and the extent of lifespan, it also has compounds that might cause adverse effects on human health such as methanol and acetaldehyde. The aim of this study was to determine the effect of time, temperature, and pectic enzymes over wine methanol and acetaldehyde concentrations during vinification. Three temperatures (20, 30, and 35 °C) and three pectic enzyme concentrations (0, 9, and 18 mL/Kg) were tested, letting fermentation to stop due to sugar depletion. Both, metanol and acetaldehyde were quantified throughout the fermentation process. Temperature reduced metanol production, observing the lowest metanol concentration (53.543 ± 3.267 mg/100 mL of wine) at 35 °C in the absence of pectic enzyme. Acetaldehyde was not affected by these variables. Alcohol, metanol, and acetaldehyde concentrations were adjusted to mathematical models with high correlations.

AbstractObjective: In this study, bioethanol production on cheese whey (prehydrolysed enzymatically using β galactosidase by isolated yeast strains was investigated.Methods: The yeast strains were isolated from Algerian natural sources (soil and grape) and further were selected on the basis of high ethanol tolerance and high ethanol production on prehydrolysed cheese whey. The selected ones were identified by morphological, physiological and biochemical characteristics. Then, a molecular identification was carried out by amplification and sequencing the D1/D2 domain of 26S rDNA region. In addition, the operating parameters of fermentation such as temperature, pH and substrate concentration (mixture of glucose and galactose) were tested for efficient yeast strain.Results: Among the selected and identified yeast strains, three strain isolates were found to be able to produce bioethanol. These strains are Hanseniaspora opuntiae Z087A0VS, Candida tropicalis Z087B0VS and Candida tropicalis Z087D0VS with an identity of 99% and 100% respectively comparing with the stocked strains in data bank. Furthermore, Hanseniaspora opuntiae presents an ethanol tolerance ethanol up to 11% whereas the two other strains of Candida tolerate up to 12%. The fermentation parameters of most efficient strain were optimized, the temperature 30°C, pH 5 and sugar concentration (glucose and galactose) of 12.5% (w/v) are considered as optimum values for Candida tropicalis Z087B0VS.Conclusion: Candida tropicalis Z087B0VS can be considered as a good candidature for industrial bioethanol production.

Aims: A better understanding of the relationship between weather conditions and wine quality would provide tools for assessing the impact of climate change and the potential for adaptation. Most studies rely on assessing wine quality by the price per bottle or by an overall ranking and then establishing general relations to weather conditions. However, such an approach may imply the addition of bias by variable winemaking techniques overcoming vintage effects. The aim of our study was therefore to implement a controlled conditions approach using grape samples from a single vineyard and a standardized micro-scale winemaking technique to produce wines in similar conditions for each vintage over more than a decade. We hope that this data will allow new insights into responses to climatic differences.Methods and results: From 2005 to 2015, data was collected from a vineyard of Hochschule Geisenheim University planted with Vitis vinifera L. cv. Pinot Noir grafted on rootstock SO4 in four field replicates. Weather conditions were recorded together with the major phenological stages, yield, infection of the bunches by Botrytis cinerea bunch rot, and pruning weight. Key primary juice compounds were analyzed and berry phenolics in skins and seeds were determined before harvest. Micro-scale winemaking was developed to produce wines in standardized conditions. The repeatability of the method to assess the extraction of anthocyanins and tannins was shown to be 2–10% and 8–12%, respectively, depending on grape maturity stage. Sugar accumulation was coupled to warmer conditions during the maturation period, and high temperatures after véraison decreased the concentration of malic acid in the juice. The accumulation of primary amino acids (N-OPA) in the juices seemed positively related to warmer conditions between bud break and flowering. Increased temperature, especially before véraison, accompanied by a lack of precipitation was related to an accumulation of tannins in fruit and wine, with a higher accumulation in skins than seeds. The temperature-sensitive anthocyanin accumulation in grapes was coupled to warmer conditions after véraison. These differences in anthocyanin concentration could also be observed in the wine.Conclusions: High-quality vintages were linked to warmer than normal growing seasons and below normal precipitation.Significance and impact of the study: The use of a micro-scale winemaking technique represents an innovative tool to provide detailed information in a controlled and reproducible way. A better understanding of the interaction between weather conditions and berry/wine compounds will help with developing improved winemaking techniques and better adapting to future impacts of climate change.

Introduction In November 2020, upon learning that the company’s Covid-19 vaccine trial had been successful, the head of Pfizer’s Vaccine Research and Development, Kathrin Jansen, celebrated with champagne – “some really good stuff” (Cohen). Bubbles seem to go naturally with celebration, and champagne is fundamentally associated with bubbles. Yet, until the late-seventeenth century, champagne was a still wine, and it only reached the familiar levels of bubbliness in the late-nineteenth century (Harding). During this period and on into the early twentieth century, “champagne” was in many ways created, defined, and defended. A “champagne bubble” was created, within which the “nature” of champagne was contested and constructed. Champagne today is the result of hundreds of years of labour by many sorts of bubble-makers: those who make the bubbly drink, and those who construct, maintain, and defend the champagne bubble. In this article, I explore some elements of the champagne bubble, in order to understand both its fragility and rigidity over the years and today. Creating the Champagne Bubble – the Labour of Centuries It is difficult to separate the physical from the mythical as regards champagne. Therefore the categorisations below are always overlapping, and embedded in legal, political, economic, and socio-cultural factors. Just as assemblage – the mixing of wine from different grapes – is an essential element of champagne wine, the champagne bubble may be called heterogeneous assemblage. Indeed, the champagne bubble, as we will see below, is a myriad of different sorts of bubbles, such as terroir, appellation, myth and brand. And just as any assemblage, its heterogeneous elements exist and operate in relation to each other. Therefore the “champagne bubble” discussed here is both one and many, all of its elements fundamentally interconnected, constituting that “one” known as “champagne”. It is not my intention to be comprehensive of all the elements, historical and contemporary. Indeed, that would not be possible within such a short article. Instead, I seek to demonstrate some of the complexity of the champagne bubble, noting the elaborate labour that has gone into its creation. The Physical Champagne and Champagne – from Soil to Bubbles Champagne means both a legally protected geographical area (Champagne), and the wine (here: champagne) produced in this area from grapes defined as acceptable: most importantly pinot noir, pinot meunier (“black” grapes), and chardonnay (“white” grape). The method of production, too, is regulated and legally protected: méthode champenoise. Although the same method is used in numerous locations, these must be called something different: metodo classico (Italy), método tradicional (Spain), Methode Cap Classique (South Africa). The geographical area of Champagne was first legally defined in 1908, when it only included the areas of Marne and Aisne, leaving out, most importantly, the area of Aube. This decision led to severe unrest and riots, as the Aube vignerons revolted in 1911, forcing the inclusion of “zone 2”: Aube, Haute-Marne, and Seine-et-Marne (Guy). Behind these regulations was a surge in fraudulent production in the early twentieth century, as well as falling wine prices resulting from increasing supply of cheap wines (Colman 18). These first appellations d’origine had many consequences – they proved financially beneficial for the “zone 1”, but less so for the “zone 2”. When both these areas were brought under the same appellation in 1927, the financial benefits were more limited – but this may have been due to the Great Depression triggered in 1929 (Haeck et al.). It is a long-standing belief that the soil and climate of Champagne are key contributors to the quality of champagne wines, said to be due to “conditions … most suitable for making this type of wine” (Simon 11). Already in the end of the nineteenth century, the editor of Vigneron champenois attributed champagne’s quality to “a fortunate combination of … chalky soil … [and] unrivalled exposure [to the sun]” (Guy 119) among other things. Factors such as soil and climate, commonly included in and expressed through the idea of terroir, undoubtedly influence grapes and wines made thereof, but the extent remains unproven. Indeed, terroir itself is a very contested concept (Teil; Inglis and Almila). It is also the case that climate change has had, and will continue to have, devastating effects on wine production in many areas, while benefiting others. The highly successful English sparkling wine production, drawing upon know-how from the Champagne area, has been enabled by the warming climate (Inglis), while Champagne itself is at risk of becoming too hot (Robinson). Champagne is made through a process more complicated than most wines. I present here the bare bones of it, to illustrate the many challenges that had to be overcome to enable its production in the scale we see today. Freshly picked grapes are first pressed and the juice is fermented. Grape juice contains natural yeasts and therefore will ferment spontaneously, but fermentation can also be started with artificial yeasts. In fermentation, alcohol and carbon dioxide (CO2) are formed, but the latter usually escapes the liquid. The secret of champagne is its second fermentation, which happens in bottles, after wines from different grapes and/or vineyards have been blended for desired characteristics (assemblage). For the second fermentation, yeast and sugar are added. As the fermentation happens inside a bottle, the CO2 that is created does not escape, but dissolves into the wine. The average pressure inside a champagne bottle in serving temperature is around 5 bar – 5 times the pressure outside the bottle (Liger-Belair et al.). The obvious challenge this method poses has to do with managing the pressure. Exploding bottles used to be a common problem, and the manner of sealing bottles was not very developed, either. Seventeenth-century developments in bottle-making, and using corks to seal bottles, enabled sparkling wines to be produced in the first place (Leszczyńska; Phillips 137). Still today, champagne comes in heavy-bottomed bottles, sealed with characteristically shaped cork, which is secured with a wire cage known as muselet. Scientific innovations, such as calculating the ideal amount of sugar for the second fermentation in 1836, also helped to control the amount of gas formed during the second fermentation, thus making the behaviour of the wine more predictable (Leszczyńska 265). Champagne is characteristically a “manufactured” wine, as it involves several steps of interference, from assemblage to dosage – sugar added for flavour to most champagnes after the second fermentation (although there are also zero dosage champagnes). This lends champagne particularly suitable for branding, as it is possible to make the wine taste the same year after year, harvest after harvest, and thus create a distinctive and recognisable house style. It is also possible to make champagnes for different tastes. During the nineteenth century, champagnes of different dosage were made for different markets – the driest for the British, the sweetest for the Russians (Harding). Bubbles are probably the most striking characteristic of champagne, and they are enabled by the complicated factors described above. But they are also formed when the champagne is poured in a glass. Natural impurities on the surface of the glass provide channels through which the gas pockets trapped in the wine can release themselves, forming strains of rising bubbles (Liger-Belair et al.). Champagne glasses have for centuries differed from other wine glasses, often for aesthetic reasons (Harding). The bubbles seem to do more than give people aesthetic pleasure and sensory experiences. It is often claimed that champagne makes you drunk faster than other drinks would, and there is, indeed, some (limited) research showing that this may well be the case (Roberts and Robinson; Ridout et al.). The Mythical Champagne – from Dom Pérignon to Modern Wonders Just as the bubbles in a champagne glass are influenced by numerous forces, so the metaphorical champagne bubble is subject to complex influences. Myth-creation is one of the most significant of these. The origin of champagne as sparkling wine is embedded in the myth of Dom Pérignon of Hautvillers monastery (1638–1715), who according to the legend would have accidentally developed the bubbles, and then enthusiastically exclaimed “I am drinking the stars!” (Phillips 138). In reality, bubbles are a natural phenomenon provoked by winter temperatures deactivating the fermenting yeasts, and spring again reactivating them. The myth of Dom Pérignon was first established in the nineteenth century and quickly embraced by the champagne industry. In 1937, Moët et Chandon launched a premium champagne called Dom Pérignon, which enjoys high reputation until this day (Phillips). The champagne industry has been active in managing associations connected with champagne since the nineteenth century. Sparkling champagnes had already enjoyed fashionability in the later seventeenth and early eighteenth century, both in the French Court, and amongst the British higher classes. In the second half of the nineteenth century, champagne found ever increasing markets abroad, and the clientele was not aristocratic anymore. Before the 1860s, champagne’s association was with high status celebration, as well as sexual activity and seduction (Harding; Rokka). As the century went on, and champagne sales radically increased, associations with “modernity” were added: “hot-air balloons, towering steamships, transcontinental trains, cars, sports, and other ‘modern’ wonders were often featured in quickly proliferating champagne advertising” (Rokka 280). During this time, champagne grew both drier and more sparkling, following consumer tastes (Harding). Champagne’s most important markets in later nineteenth century included the UK, where the growing middle classes consumed champagne for both celebration and hospitality (Harding), the US, where (upper) middle-class women were served champagne in new kinds of consumer environments (Smith; Remus), and Russia, where the upper classes enjoyed sweeter champagne – until the Revolution (Phillips 296). The champagne industry quickly embraced the new middle classes in possession of increasing wealth, as well as new methods of advertising and marketing. What is remarkable is that they managed to integrate enormously varied cultural thematics and still retain associations with aristocracy and luxury, while producing and selling wine in industrial scale (Harding; Rokka). This is still true today: champagne retains a reputation of prestige, despite large-scale branding, production, and marketing. Maintaining and Defending the Bubble: Formulas, Rappers, and the Absolutely Fabulous Tipplers The falling wine prices and increasing counterfeit wines coincided with Europe’s phylloxera crisis – the pest accidentally brought over from North America that almost wiped out all Europe’s vineyards. The pest moved through Champagne in the 1890s, killing vines and devastating vignerons (Campbell). The Syndicat du Commerce des vins de Champagne had already been formed in 1882 (Rokka 280). Now unions were formed to fight phylloxera, such as the Association Viticole Champenoise in 1898. The 1904 Fédération Syndicale des Vignerons was formed to lobby the government to protect the name of Champagne (Leszczyńska 266) – successfully, as we have seen above. The financial benefits from appellations were certainly welcome, but short-lived. World War I treated Champagne harshly, with battle lines stuck through the area for years (Guy 187). The battle went on also in the lobbying front. In 1935, a new appellation regime was brought into law, which came to be the basis for all European systems, and the Comité National des appellations d'origine (CNAO) was founded (Colman 1922). Champagne’s protection became increasingly international, and continues to be so today under EU law and trade deals (European Commission). The post-war recovery of champagne relied on strategies used already in the “golden years” – marketing and lobbying. Advertising continued to embrace “luxury, celebration, transport (extending from air travel to the increasingly popular automobile), modernity, sports” (Guy 188). Such advertisement must have responded accurately to the mood of post-war, pre-depression Europe. Even in the prohibition US it was known that the “frivolous” French women might go as far as bathe in champagne, like the popular actress Mistinguett (Young 63). Curiously, in the 1930s Soviet Russia, “champagne” (not produced in Champagne) was declared a sign of good living, symbolising the standard of living that any Soviet worker had access to (at least in theory) (Gronow). Today, the reputation of champagne is fiercely defended in legal terms. This is not only in terms of protection against other sparkling wine making areas, but also in terms of exploitation of champagne’s reputation by actors in other commercial fields, and even against mass market products containing genuine champagne (Mahy and d’Ath; Schneider and Nam). At the same time, champagne has been widely “democratised” by mass production, enabled partly by increasing mechanisation and scientification of champagne production from the 1950s onwards (Leszczyńska 266). Yet champagne retains its association with prestige, luxury, and even royalty. This has required some serious adaptation and flexibility. In what follows, I look into three cultural phenomena that illuminate processes of such adaptation: Formula One (F1) champagne spraying, the 1990s sitcom Absolutely Fabulous, and the Cristal racism scandal in 2006. The first champagne bottle is said to have been presented to F1 grand prix winner in Champagne in 1950 (Wheels24). Such a gesture would have been fully in line with champagne’s association with cars, sport, and modernity. But what about the spraying? Surely that is not in line with the prestige of the wine? The first spraying is attributed to Jo Siffert in 1966 and Dan Gurney in 1967, the former described as accidental, the latter as a spontaneous gesture of celebration (Wheels24; Dobie). Moët had become the official supplier of F1 champagnes in 1966, and there are no signs that the new custom would have been problematic for them, as their sponsorship continued until 1999, after which Mumm sponsored the sport for 15 years. Today, the champagne to be popped and sprayed is Chanson, in special bottles “coated in the same carbon fibre that F1 cars are made of” (Wheels24). Such an iconic status has the spraying gained that it features in practically all TV broadcasts concerning F1, although non-alcoholic substitute is used in countries where sale of alcohol is banned (Barker et al., “Quantifying”; Barker et al., “Alcohol”). As disturbing as the champagne spraying might look for a wine snob, it is perfectly in line with champagne’s marketing history and entrepreneurial spirit shown since the nineteenth century. Nor is it unheard of to let champagne spray. The “art” of sabrage, opening champagne bottle with a sable, associated with glamour, spectacle, and myth – its origin is attributed to Napoleon and his officers – is perfectly acceptable even for the snob. Sparkling champagne was always bound up with joy and celebration, not a solemn drink, and the champagne bubble was able to accommodate middle classes as well as aristocrats. This brings us to our second example, the British sitcom Absolutely Fabulous. The show, first released in 1992, featured two women, “Eddy” (Jennifer Saunders) and “Patsy” (Joanna Lumley), who spent their time happily smoking, taking drugs, and drinking large quantities of “Bolly” (among other things). Bollinger champagne may have initially experienced “a bit of a shock” for being thus addressed, but soon came to see the benefits of fame (French). In 2005, they hired PR support to make better use of the brand’s “Ab Fab” recognisability, and to improve its prestige reputation in order to justify their higher price range (Cann). Saunders and Lumley were warmly welcomed by the Bollinger house when filming for their champagne tour Absolutely Champers (2017). It is befitting indeed that such controversial fame came from the UK, the first country to discover sparkling champagne outside France (Simon 48), and where the aspirational middle classes were keen to consume it already in the nineteenth century (Harding). More controversial still is the case of Cristal (made by Louis Roederer) and the US rap world. Enthusiastically embraced by the “bling-bling” world of (black) rappers, champagne seems to fit their ethos well. Cristal was long favoured as both a drink and a word in rap lyrics. But in 2006, the newly appointed managing director at the family owned Roederer, Frédéric Rouzaud, made comments considered racist by many (Woodland). Rouzard told in an interview with The Economist that the house observed the Cristal-rap association “with curiosity and serenity”. He reportedly continued: “but what can we do? We can’t forbid people from buying it. I’m sure Dom Pérignon or Krug would be delighted to have their business”. It was indeed those two brands that the rapper Jay-Z replaced Cristal with, when calling for a boycott on Cristal. It would be easy to dismiss Rouzard’s comments as snobbery, or indeed as racism, but they merit some more reflection. Cristal is the premium wine of a house that otherwise does not enjoy high recognisability. While champagne’s history involves embracing new sorts of clientele, and marketing flexibly to as many consumer groups as possible (Rokka), this was the first spectacular crossing of racial boundaries. It was always the case that different houses and their different champagnes were targeted at different clienteles, and it is apparent that Cristal was not targeted at black rap artists. Whereas Bollinger was able to turn into a victory the questionable fame brought by the white middle-class association of Absolutely Fabulous, the more prestigious Cristal considered the attention of the black rapper world more threatening and acted accordingly. They sought to defend their own brand bubble, not the larger champagne bubble. Cristal’s reputation seems to have suffered little – its 2008 vintage, launched in 2018, was the most traded wine of that year (Schultz). Jay-Z’s purchase of his own champagne brand (Armand de Brignac, nicknamed Ace of Spades) has been less successful reputation-wise (Greenburg). It is difficult to break the champagne bubble, and it may be equally difficult to break into it. Conclusion In this article, I have looked into the various dilemmas the “bubble-makers” of Champagne encountered when fabricating what is today known as “champagne”. There have been moments of threat to the bubble they formed, such as in the turn of nineteenth and twentieth centuries, and eras of incomparable success, such as from the 1860s to 1880s. The discussion has demonstrated the remarkable flexibility with which the makers and defenders of champagne have responded to challenges, and dealt with material, socio-cultural, economic, and other problems. It feels appropriate to end with a note on the current challenge the champagne industry faces: Covid-19. The pandemic hit champagne sales exceptionally hard, leaving around 100 million bottles unsold (Micallef). This was not very surprising, given the closure of champagne-selling venues, banning of public and private celebrations, and a general mood not particularly prone to (or even likely to frown upon) such light-hearted matters as glamour and champagne. Champagne has survived many dramatic drops in sales during the twentieth century, such as the Great Depression of the 1930s, and the post-financial crisis collapse in 2009. Yet they seem to be able to make astonishing recoveries. Already, there are indicators that many people consumed more champagne during the festive end-of-year season than in previous years (Smithers). For the moment, it looks like the champagne bubble, despite its seeming fragility, is practically indestructible, no matter how much its elements may suffer under various pressures and challenges. References Barker, Alexander, Magdalena Opazo-Breton, Emily Thomson, John Britton, Bruce Granti-Braham, and Rachael L. Murray. “Quantifying Alcohol Audio-Visual Content in UK Broadcasts of the 2018 Formula 1 Championship: A Content Analysis and Population Exposure.” BMJ Open 10 (2020): e037035. <https://bmjopen.bmj.com/content/10/8/e037035>. Barker, Alexander B., John Britton, Bruce Grant-Braham, and Rachael L. Murray. “Alcohol Audio-Visual Content in Formula 1 Television Broadcasting.” BMC Public Health 18 (2018): 1155. <https://bmcpublichealth.biomedcentral.com/articles/10.1186/s12889-018-6068-3>. Campbell, Christy. Phylloxera: How Wine Was Saved for the World. London: Harper, 2004. Cann, Richard. “Bolllinger Signs Agency to Reclaim Ab Fab Status.” PR Week 4 Mar. 2005. 4 Mar. 2021 <https://www.prweek.com/article/472221/bollinger-signs-agency-reclaim-ab-fab-status>. Cohen, Jon. “Champagne and Questions Greet First Data Showing That a COVID-19 Vaccine Works.” Science 9 Nov. 2020. 4 Mar. 2021 <https://www.sciencemag.org/news/2020/11/champagne-and-questions-greet-first-data-showing-covid-19-vaccine-works>. Colman, Tyler. Wine Politics: How Governments, Environmentalists, Mobsters, and Critics Influence the Wines We Drink. Berkeley: University of California Press, 2008. Dobie, Stephen. “The Story of Motorsport’s First Ever Champagne Spray.” TopGear 15 Jan. 2018. 4 Mar. 2021 <https://www.topgear.com/car-news/motorsport/story-motorsports-first-ever-champagne-spray>. European Commission. “Wine.” 4 Mar. 2021 <https://ec.europa.eu/info/food-farming-fisheries/plants-and-plant-products/plant-products/wine_en#:~:text=Related%20links-,Overview,consumption%20and%2070%25%20of%20exports>. French, Phoebe. “Joanna Lumley and Jennifer Saunders to Star in Absolutely Champers.” The Drinks Business 20 Dec. 2017. 4 Mar. 2021 <https://www.thedrinksbusiness.com/2017/12/joanna-lumley-and-jennifer-saunders-to-star-in-absolutely-champers/>. Greenburg, Zack O. “The Real Story behind Jay Z's Champagne Deal.” Forbes 6 Nov. 2014. 4 Mar. 2021 <https://www.forbes.com/sites/zackomalleygreenburg/2014/11/06/why-jay-zs-champagne-news-isnt-so-new/?sh=6e4eb8f07528>. Gronow, Jukka. “Caviar with Champagne Good Life and Common Luxury in Stalin's Soviet Union.” Suomen Antropologi 4 (1998). Guy, Colleen M. When Champagne Became French: Wine and the Making of a National Identity. London: Johns Hopkins University Press, 2003. Haeck, Catherine, Giulia Meloni, and Johan Swinnen. “The Value of Terroir: A Historical Analysis of the Bordeaux and Champagne Geographical Indications.” Applied Economic Perspectives and Policy 41.4 (2019): 598–619. <https://onlinelibrary.wiley.com/doi/abs/10.1093/aepp/ppz026>. Harding, Graham. “The Making of Modern Champagne: How and Why the Taste for and the Taste of Champagne Changed in Nineteenth Century Britain.” Consumption Markets & Culture 42.1 (2021): 6-29. <https://www.tandfonline.com/doi/abs/10.1080/10253866.2020.1713765?journalCode=gcmc20>. Inglis, David. “Wine Globalization: Longer-Term Dynamics and Contemporary Patterns.” The Globalization of Wine. Eds. David Inglis and Anna-Mari Almila. London: Bloomsbury, 2019. 21-46. Inglis, David, and Anna-Mari Almila. “Introduction: The Travels and Tendencies of Wine.” The Globalization of Wine. Eds. David Inglis and Anna-Mari Almila. London: Bloomsbury, 2019. 1-20. Leszczyńska, D. “A Cluster and Its Trajectory: Evidence from the History of the French Champagne Production Cluster.” Labor History 57.2 (2016): 258-276. <https://www.tandfonline.com/doi/abs/10.1080/0023656X.2016.1161140>. Liger-Belair, Gérard, Guillaume Polidori, and Philippe Jeandet. “Recent Advances in the Science of Champagne Bubbles.” Chemical Society Reviews 37 (2008): 2490–2511. <https://pubs.rsc.org/en/content/articlelanding/2008/cs/b717798b#!divAbstract>. Mahy, Aude, and Florence d’Ath. “The Case of the ‘Champagner Sorbet’ – Unlawful Exploitation or Legitimate Use of the Protected Name ‘Champagne’?” EFFL 1 (2017): 43-48. <https://www.jstor.org/stable/26451418?seq=1>. Micallef, Joseph V. “How Champagne Is Bouncing Back after the COVID-19 Pandemic.” Forbes 15 Nov. 2020. 4 Mar. 2021 <https://www.forbes.com/sites/joemicallef/2020/11/15/how-champagne-is-bouncing-back-after-the-covid-19-pandemic/?sh=3300e4125784>. Phillips, Rod. A Short History of Wine. London: Penguin, 2000. Remus, Emily A. “Tippling Ladies and the Making of Consumer Culture: Gender and Public Space in ‘Fin-de- Siècle’ Chicago.” The Journal of American History 101.3 (2014): 751-77. <https://academic.oup.com/jah/article/101/3/751/796447?login=true>. Ridout, Fran, Stuart Gould, Carlo Nunes, and Ian Hindmarch. “The Effects of Carbon Dioxide in Champagne on Psychometric Performance and Blood-Alcohol Concentration.” Alcohol and Alcoholism 38.4 (2003): 381-85. <https://academic.oup.com/alcalc/article/38/4/381/232628>. Roberts, C., and S.P. Robinson. “Alcohol Concentration and Carbonation of Drinks: The Effect on Blood Alcohol Levels.” Journal of Forensic and Legal Medicine 14.7 (2007): 398-405. <https://pubmed.ncbi.nlm.nih.gov/17720590/>. Robinson, Frances. “Champagne Will Be Too Hot for Champagne Research Warns.” Decanter. 12 Jan. 2004. 4 Mar. 2021 <https://www.decanter.com/wine-news/champagne-will-be-too-hot-for-champagne-research-warns-103258/>. Rokka, Joonas. “Champagne: Marketplace Icon.” Consumption Markets & Culture 20.3 (2017): 275-283. <https://www.tandfonline.com/doi/abs/10.1080/10253866.2016.1177990?journalCode=gcmc20>. Schneider, Marius, and Nora Ho Tu Nam. “Champagne Makes the Dough Sour: EUIPO Board of Appeal Allows Opposition against Registration of Champagnola Trade Mark Based on Evocation of Champagne PDO.” Journal of Intellectual Property Law & Practice 15.9 (2020): 675-676. <https://academic.oup.com/jiplp/article/15/9/675/5905791>. Schultz, Abby. “20 Minutes With: Frédéric Rouzaud on Cristal, Biodynamics, and Zero Dosage.” Penta. 31 Dec. 2018. 4 Mar. 2021 <https://www.barrons.com/articles/20-minutes-with-frederic-rouzaud-on-cristal-biodynamics-and-zero-dosage-01546280265>. Simon, André L. The History of Champagne. London: Octobus, 1972. Smith, Andrew F. Drinking History: Fifteen Turning Points in the Making of American Beverages. New York: Columbia University Press, 2013. Smithers, Rebecca. “Britons Turn to Luxury Food and Drink to See Out Dismal 2020 in Style.” The Guardian 28 Dec. 2020. 4 Mar. 2021 <https://www.theguardian.com/business/2020/dec/28/britons-turn-luxury-food-drink-see-out-dismal-2020-style?CMP=Share_AndroidApp_Gmail>. Teil, Geneviève. “No Such Thing as Terroir? Objectivities and the Regimes of Existence of Objects.” Science, Technology & Human Values 37.5 (2012): 478-505. <https://journals.sagepub.com/doi/abs/10.1177/0162243911423843>. Wheels24. “Champagne Returns to F1 podium.” 2 Aug. 2017. 4 Mar. 2021 <https://www.news24.com/wheels/FormulaOne/champagne-returns-to-f1-podium-20170802>. Woodland, Richard. “Rapper Jay-Z Boycotts ‘Racist’ Cristal.” Decanter 16 June 2006. 4 Mar. 2021 <https://www.decanter.com/wine-news/rapper-jay-z-boycotts-racist-cristal-94054/>. Young, Robert K. “Out of the Ashes: The American Press and France's Postwar Recovery in the 1920s.” Historical Reflections / Réflexions Historiques 28.1 (2002): 51-72. <https://www.jstor.org/stable/41299224?seq=1>.