Commercial Active Dry Yeast Research Articles

EVOLUTION OF CIS AND TRANS-RESVERATROL IN WINES OBTAINED THROUGH INOCULATION WITH ACTIVE DRY YEAST AT PIETROASA VINEYARD

This paper presents the results of the testing during the winemaking process using commercial active dry yeasts (CADY) in parallel with yeast biomass produced starting from yeasts isolated from Pietroasa area (Romania) (PAY). PAYs were obtained based on autochthonous yeasts isolated from Busuioacă de Bohotin (BB) and Tămâioasa Românească (TR) grape varieties. Two variants of must for each grape variety were taken in study, i.e. obtained from early harvested grapes (TRD and BBD) and from late harvested grapes (TRS and BBS). Winemaking experiments were carried out with 7 types of commercial yeasts and two types of commercial nutrients. The alcoholic concentration of the experimental wines varied from 11.8 % for WTRD to 18.4% for WBBD. The WBBD & S samples showed a higher average trans-resveratrol content (1.716 and 1.555 mg/L, respectively) compared to the WTRD & S wine samples (0.590 and 1.365 mg/L, respectively). The variation in the cis-resveratrol content between the experimental variants produced with PAY was not significant.

Effects of Saccharomyces cerevisiae boulardii (CNCM I-1079) on feed intake, blood parameters, and production during early lactation.

The periparturient period is a metabolically demanding time for dairy animals because of increased nutrient requirements for milk yield. The objective of this study was to investigate the effect of feeding Saccharomyces cerevisiae boulardii (CNCM I-1079), a commercial active dry yeast (ADY), in dairy cows on productive and metabolic measures during the periparturient period. Primiparous (n = 33) and multiparous (n = 35) cows were fed a close-up total mixed ration (TMR) before calving and a lactation TMR postpartum. Three weeks before expected calving time, animals were blocked by parity and body weight and then randomly assigned to either control group (control; n = 34) or treatment (ADY; n = 34). All animals were housed in a tie-stall barn with individual feed bunks; the ADY animals received supplementary Saccharomyces cerevisiae boulardii (CNCM I-1079), top dressed daily at a predicted dosage of 1.0 × 1010 cfu (12.5 g) per head. Blood samples were collected weekly along with milk yield and milk composition data; feed intake data were collected daily. Serum samples were analyzed for glucose, nonesterified fatty acid, β-hydroxybutyrate, haptoglobin (Hp), and the cytokines tumor necrosis factor-α, IL-6, and IL-18. Colostrum samples collected within the first 6 to 10 h were analyzed for somatic cell score and IgG, IgA, and IgM concentrations. Data were analyzed using PROC GLIMMIX in SAS with time as a repeated measure; model included time, parity, treatment, and their interactions. The ADY groups had greater milk yield (39.0 ± 2.4 vs. 36.7 ± 2.3 kg/d) and tended to produce more energy-corrected milk with better feed efficiency. There was no difference in plasma glucose, serum nonesterified fatty acid, serum β-hydroxybutyrate, Hp, IL-6, or IL-18 due to ADY treatment. The tumor necrosis factor-α increased in ADY-supplemented animals (1.17 ± 0.69 vs. 4.96 ± 7.7 ng/mL), though week, parity, and their interactions had no effect. Serum amyloid A tended to increase in ADY-supplemented animals when compared to control animals and was additionally affected by week and parity; there were no significant interactions. No difference in colostrum IgG, IgA, and IgM was observed between treatments. Supplementing transition cow TMR with ADY (CNCM I-1079) improved milk production and tended to improve efficiency in early lactation; markers of inflammation were also influenced by ADY treatment, though the immunological effect was inconsistent.

Effect of Bentonite Addition to Pedro Ximénez White Grape Musts before Their Fermentation with Selected Yeasts on the Major Volatile Compounds and Polyols of Wines and Tentative Relationships with the Sensorial Evaluation.

In this work, we study the effect of bentonite addition to the grape must before alcoholic fermentation on the chemical composition and sensorial profile of the obtained wines. Fermentations were carried out with two Saccharomyces cerevisiae commercial active dry yeasts treated or not with bentonite and were compared with a control wine obtained by spontaneous fermentation (using the grape must microbiota). Several significant effects on the chemical and sensorial attributes were established by statistical treatments. The selection by multiple variable analysis of seven volatile molecules (ethyl acetate; methanol; 1-propanol; isobutanol; 2-methyl-1-butanol; 3-metyl-1-butanol and 2-phenylethanol) provided several footprints that provide an easy visualization of bentonite effects on wine volatile compounds. A Principal Component Analysis carried out with all the compounds quantified by Gas-Chromatography revealed that the first two Principal Components explain 60.15 and 25.91%, respectively, of the total variance and established five groups that match with the five wines analyzed. Lastly, predictive models at p ≤ 0.05 level for the attributes sight, smell and taste were obtained by Partial Least Squared regression analysis of selected chemical variables.

Screening low-methanol and high-aroma produced yeasts for cider fermentation by transcriptive characterization.

The commercial active dry yeast strains used for cider production in China are far behind the requirements of the cider industry development in recent decades. In this study, eight yeasts, including Saccharomyces cerevisiae, Schizosaccharomyces pombe, Pichia bruneiensis, and Pichia kudriavzevii, were screened and assessed by growth performance, methanol production, aroma analysis, and their transcriptive characterization. Saccharomyces cerevisiae strains WFC-SC-071 and WFC-SC-072 were identified as promising alternatives for cider production. Strains WFC-SC-071 and WFC-SC-072 showed an excellent growth capacity characterized by 91.6 and 88.8% sugar utilization, respectively. Methanol production by both strains was below 200 mg/L. Key aroma compounds imparting cider appreciably characteristic aroma increased in cider fermented by strains WFC-SC-071 and WFC-SC-072. RT-qPCR analysis suggested that most genes associated with growth capacity, carbohydrate uptake, and aroma production were upregulated in WFC-SC-071 and WFC-SC-072. Overall, two Saccharomyces cerevisiae strains are the optimal starters for cider production to enable the diversification of cider, satisfy the differences in consumer demand, and promote cider industry development.

Survey of Inoculated Commercial Saccharomyces cerevisiae in Winery-Based Trials

Wine production has developed from spontaneous to controlled fermentations using commercial active dry yeasts (ADY). In this study, S. cerevisiae commercial ADY were tested, and yeast community dynamics were monitored at different fermentation stages in three winery-based trials with volumes ranging from 60 L to 250 hL. The differentiation of S. cerevisiae strains was achieved using microsatellite markers. In Experiment 1, results showed that both ADY strains revealed similar profiles, despite being described by the producer as having different properties. In Experiment 2, higher genetic diversity was detected when co-inoculation was tested, while in sequential inoculation, the initial ADY seemed to dominate throughout all fermentation. Pilot-scale red wine fermentations were performed in Experiment 3, where one single ADY strain was tested along with different oenological additives. Surprisingly, these trials showed an increase in distinct profiles towards the end of fermentation, indicating that the dominance of the ADY was lower than in the blank modality. The use of ADY is envisaged to promote a controlled and efficient alcoholic fermentation, and their purchase represents an important cost for wineries. Therefore, it is most relevant to survey commercial ADY during wine fermentation to understand if their use is effective.

Volatile Compounds Analysis in Blueberry Wine Fermented with Different Yeasts by Gas Chromatography-Ion Mobility Spectrometry

The volatile compounds of blueberry wine fermented with four commercial active dry yeasts (Lalvin K1, DV10, Angel ADT and UV43) were analysed by GC-IMS coupled with fingerprint information and principal component analysis. The results showed that 26 volatile substances, including some dimers, were determined from blueberry wine. Esters and alcohols were the main volatile substances, with small amounts of acids, aldehydes and ketones. Among them, isobutyl acetate, n-propyl acetate, 2-methylbutyrate, isobutyrate, propionate, ethyl formate and ethyl acetate were the main characteristic flavor substances of blueberry wine. The results of fingerprint information and principal component analysis showed that higher level of esters were found in wines fermented with K1 and DV10 yeasts, and higher level of acetoin was found in Angel ADT while higher level of acetic acid and 2-methylpropionic acid were found in UV43. GC-IMS technology showed a promising application for improving the aroma sensation and constructing the flavor fingerprint of blueberry wine products fermented by different yeasts.

Obtención de alcohol etílico a partir de dos variedades de cabuya: alternativa para su industrialización

Este proyecto de investigación estudia la utilización de plantas nativas de Furcraea cabuya var. integra y Furcraea cabuya var. cabuya del litoral ecuatoriano, cosechadas en los poblados de: Ayampe, Montecristi, Tosagua, Portoviejo, con el objetivo principal de obtener alcohol etílico mediante la fermentación, destilación y filtración y de esta forma conseguir una bebida alcohólica apta para el consumo humano. En el acondicionamiento y desarrollo experimental se consideran los diferentes parámetros como los diferentes tiempos y temperaturas de cocción, las pruebas con diferentes tipos de inóculos y concentraciones de levaduras, además de diferentes suplementaciones nutritivas (con especial atención a la fuente de nitrógeno). Estableciendo como temperatura óptima para la hidrólisis de la inulina 117 °C durante 4 horas y el proceso fermentativo óptimo utilizando levadura seca activa comercial al 1% p/v con respecto al mosto y una suplementación con sulfato de amonio de 0,15 g/100mL de mosto. El mosto de cabuya rectificado genera una bebida con exquisito sabor a Tequila, la cual luego de ser evaluada sensorialmente demostró que el licor producido se encuentra dentro de los parámetros de aceptación de los consumidores. Palabras clave: etanol, cabuya, destilación, fermentación, mosto, tequila.

The impact of active dry yeasts in commercial wineries from the Denomination of Origen "Vinos de Madrid", Spain.

This paper has studied the success of implantation for 16 commercial active dry yeasts (ADYs) during industrial fermentation (30) and the impact of these yeasts during spontaneous fermentations (19) in 10 wineries from the Denomination of Origin "Vinos de Madrid" over two consecutive years. Yeasts strains were identified by molecular techniques, pulsed field electrophoresis and microsatellite analysis. According to these techniques, all the ADYs were different with the exceptions of two strains, L2056 and Rh, which showed the same karyotype and loci size. The results showed that inoculating fermentations with ADYs did not ensure their dominance throughout the fermentation; the implantation level of ADYs was above 80% in only 9 of the 30 commercial fermentations studied; while in 16 fermentations, the dominance of the inoculated ADYs was below 50%. The type of vinification with the best implantation results overall were those associated with red wine fermentations. ADYs affected spontaneous fermentations, although their impact was observed to decrease in the second year of the study. Therefore, specific adaptation studies are necessary before using commercial yeasts during the fermentation process. At the same time, a study was carried out on the frequency of commercial strains in IMIDRA's yeast collection, made up of strains isolated from spontaneous fermentations of the different areas and cellars since the beginning of the Denomination of Origin "Vinos de Madrid" in 1990. Six different ADYs were found with a frequency of less than 5%.

Variation in content of tyramine, histamine, 2‐phenylethylamine, tryptamine and their precursor amino acids in a Chardonnay wine by using different commercial active dry yeasts and nitrogen sources

SummaryWine is a fermented beverage that could be affected by high concentrations of biogenic amines, thus altering organoleptic and health properties. In this work, the capacity of different selected yeast strains regarding to affect the content of tyramine, histamine, 2‐phenylethylamine, tryptamine and their precursor amino acids (pAA) during fermentation has been described. Overall, biogenic amines (BAs) and pAA contents appeared significantly related both to the strain and to the nutrient supplementation applied. Among BAs, without and (with) nitrogen source addition, the concentrations covered a range between 0.1 and 9.5 (0.1–10.3) mg L−1 for tyramine; 0.1 and 4.4 (0.1–4.6) mg L−1 for histamine; 0.4 and 1.1 (0.4–1.4) mg L−1 for 2‐phenylethylamine; and 0.02 and 0.14 (0.01–0.12) mg L−1 for tryptamine, respectively. Considering that also wine yeast species are responsible for BAs formation, the evidence from fermentation trials underlined the relevance of a careful choice of ADY strains and nitrogen sources in the management of alcoholic fermentation in wine.

Inoculation strategies to improve persistence and implantation of commercial S. cerevisiae strains in red wines produced with prefermentative cold soak

Prefermentative cold soak is a winemaking technique aimed at enhancing aroma and colour extraction in red wines. This study aimed at evaluating implantation and persistence rates of commercial active dry yeast strains (ADY) in wines produced with cold soak using two different inoculation strategies. Cold soak was conducted at 4 ± 1 °C, 8 ± 1 °C and 12 ± 1 °C for 7 days. Two yeast strains (Lalvin ICV D254 and Lalvin Rhône 2056) were inoculated before and after cold soak. Implantation of Lalvin ICV D254 at the end of cold soak was higher when conducted at 8 °C and 4 °C, whereas it was undetectable (<5%) at 12 °C. Lalvin Rhône 2056 showed implantation percentages at the end of cold soak ranging from 40% to 100%, with higher percentages in cold soak conducted at 12 °C. Moderate persistence of ADY at the end of alcoholic fermentation was observed in all treatment inoculated before cold soak. In the treatments inoculated after cold soak, the persistence of Lalvin ICV D254 ranged from 12 to 38% whereas Lalvin Rhône 2056 was less than 5% at the end of alcoholic fermentation. Overall, persistence of both strains improved in inoculation before cold soak relative to inoculation after cold soak.

Influence of different commercial active dry yeasts on histaminol production during wine alcoholic fermentation

AbstractThe performance of different selected yeast strains regarding the capacity to convert histidine into histaminol during fermentation has been described and discussed in a model grape must (Chardonnay). First, the activity of 10 commercial active dry yeast strains (ADY) in the native grape must both with and without a nitrogen supplementation has been compared. In the second set of fermentations, after selecting the four best‐performing strains, their performances using four different nitrogen sources have been tested. During fermentation process it was not possible to identify a common trend. In some cases, histaminol content appeared related to the nutrient supplementation, however, in other cases a different behaviour emerged. However, as shown by the experimental evidences, among the two variables considered the nature of yeast strain was slightly prevalent.

Composition ofSaccharomyces cerevisiaestrains in spontaneous fermentations of Pinot Noir and Chardonnay

There is a lack of knowledge about the composition of Saccharomyces cerevisiae strains in spontaneous fermentations of Pinot Noir and Chardonnay cultivars. The objectives were to determine the relative abundance of indigenous and commercial S. cerevisiae strains in spontaneous fermentations at three wineries from the two cultivars and to compare the composition of the S. cerevisiae strains between cultivars and wineries. Three fermentation vessels were sampled at three stages of fermentation for each cultivar at each winery. Isolates were identified to the strain level using seven microsatellite loci. Commercial S. cerevisiae strains were isolated at a frequency higher than that of the indigenous strains at each winery for both cultivars. The composition of S. cerevisiae strains was different for each cultivar and at each winery. Our results illustrate the clear influence of inoculated commercial active dry yeast strains on the composition of S. cerevisiae strains in spontaneous fermentations at wineries conducting both inoculated and spontaneous fermentations.

Oxygen Consumption by Postfermentation Wine Yeast Lees: Factors Affecting Its Rate and Extent under Oenological Conditions.

Postfermentation wine yeast lees show antioxidant properties based on their ability to consume dissolved oxygen. The oxygen consumption capacity of suspended yeast lees obtained after fermentations with six commercial active dry yeast strains was investigated in model, white and red wines using fluorescence-based oxygen sensors operating in a nondestructive way. In model solution, the oxygen consumption rate of yeast lees was shown to depend on their amount, yeast strain, sulfur dioxide and temperature. It is slightly lower in red than in white wines. It is strongly decreased by current levels of free sulfur dioxide, thus excluding the complementary use of both as antioxidants in wine. However, in 25 randomly sampled white wines produced under commercial conditions, the rate and extent of oxygen consumption during the first six months of postfermentation had no significant correlation with any of these interacting factors, making it difficult to predict the actual antioxidant effect of yeast lees. In these wines, yeast lees consumed 0 to 47% of the dissolved oxygen. Although total oxygen consumption capacity of yeast lees is not a limiting factor under commercial winemaking conditions, their oxygen consumption proceeds at a limited rate that reduces but cannot totally prevent concomitant chemical oxidation of the wine.

Impact of different spray-drying conditions on the viability of wine Saccharomyces cerevisiae strains.

Spray-drying (SD) is widely considered a suitable method to preserve microorganisms, but data regarding yeasts are still scanty. In this study, the effect of growing media, process variables and carriers over viability of a wild wine Saccharomyces (S.) cerevisiae LM52 was evaluated. For biomass production, the strain was grown (batch and fed-batch fermentation) in a synthetic, as well as in a beet sugar molasses based-medium. Drying of cells resuspended in several combinations of soluble starch and maltose was performed at different inlet and outlet temperatures. Under the best conditions-suspension in soluble starch plus maltose couplet to inlet and outlet temperatures of 110 and 55 °C, respectively-the loss of viability of S. cerevisiae LM52 was 0.8 ± 0.1 and 0.5 ± 0.2 Log c.f.u. g(-1) for synthetic and molasses-based medium, respectively. Similar results were obtained when S. cerevisiae strains Zymoflore F15 and EC1118, isolated from commercial active dry yeast (ADY), were tested. Moreover, powders retained a high vitality and showed good fermentation performances up to 6 month of storage, at both 4 and -20 °C. Finally, fermentation performances of different kinds of dried formulates (SD and ADY) compared with fresh cultures did not show significant differences. The procedure proposed allowed a small-scale production of yeast in continuous operation with relatively simple equipment, and may thus represent a rapid response-on-demand for the production of autochthonous yeasts for local wine-making.

The Occurrence of Beer Spoilage Lactic Acid Bacteria in Craft Beer Production.

Beer is one of the world's most ancient and widely consumed fermented alcoholic beverages produced with water, malted cereal grains (generally barley and wheat), hops, and yeast. Beer is considered an unfavorable substrate of growth for many microorganisms, however, there are a limited number of bacteria and yeasts, which are capable of growth and may spoil beer especially if it is not pasteurized or sterile-filtered as craft beer. The aim of this research study was to track beer spoilage lactic acid bacteria (LAB) inside a brewery and during the craft beer production process. To that end, indoor air and work surface samples, collected in the brewery under study, together with commercial active dry yeasts, exhausted yeasts, yeast pellet (obtained after mature beer centrifugation), and spoiled beers were analyzed through culture-dependent methods and PCR-DGGE in order to identify the contaminant LAB species and the source of contamination. Lactobacillus brevis was detected in a spoiled beer and in a commercial active dry yeast. Other LAB species and bacteria ascribed to Staphylococcus sp., Enterobaceriaceae, and Acetobacter sp. were found in the brewery. In conclusion, the PCR-DGGE technique coupled with the culture-dependent method was found to be a useful tool for identifying the beer spoilage bacteria and the source of contamination. The analyses carried out on raw materials, by-products, final products, and the brewery were useful for implementing a sanitization plan to be adopted in the production plant.

Membrane lipid variability in Saccharomyces cerevisiae wine strains rehydrated in the presence of metabolic activators.

Slight variations in lipid composition of wine yeast membranes can alter some essential functions including selective nutrient transport and ion permeability. The absence of oxygen during alcoholic fermentation inhibits fatty acid desaturation and sterol biosynthesis, thereby reducing the stress resistance of yeast cells. In this work, membrane lipids in two commercial active dry yeast strains rehydrated in the presence of three activators (ergosterol, tetrahydrofolic acid, and manganese) were studied. Each was assayed at three different concentrations. The effect of these activators on the phospholipid, neutral lipid, and fatty acid contents in cell membranes was assessed. Also, cell viability and fermentation kinetics were determined. Ergosterol was found to shorten the lag phase and improve cell viability and membrane lipid composition; tetrahydrofolic acid raised neutral lipid levels; and manganese(II) increased cell viability and modified phospholipid composition and linoleic acid concentration. All activators interacted with yeasts in a strain-dependent way.

Technological properties of indigenous wine yeast strains isolated from wine production regions of Turkey

The purpose of this study was to evaluate the important technological and fermentative properties of wine yeast strains previously isolated from different wine producing regions of Turkey. The determination of the following important properties was made: growth at high temperatures; fermentative capability in the presence of high sugar concentration; fermentation rate; hydrogen sulfide production; killer activity; resistance to high ethanol and sulfur dioxide; foam production; and enzymatic profiles. Ten local wine yeast strains belonging to Saccharomyces, and one commercial active dry yeast as a reference strain were evaluated. Fermentation characteristics were evaluated in terms of kinetic parameters, including ethanol yield (YP/S), biomass yield (YX/S), theoretical ethanol yield (%), specific ethanol production rate (qp; g/gh), specific glucose uptake rate (qs; g/gh), and the substrate conversion (%). All tested strains were able to grow at 37°C and to start fermentation at 30° Brix, and were resistant to high concentrations of sulfur dioxide. 60% of the strains were weak H2S producers, while the others produced high levels. Foam production was high, and no strains had killer activity. Six of the tested strains had the ability to grow and ferment at concentrations of 14% ethanol. Except for one strain, all fermented most of the media sugars at a high rate, producing 11.0-12.4% (v/v) ethanol. Although all but one strain had suitable characteristics for wine production, they possessed poor activities of glycosidase, esterase and proteinase enzymes of oenological interest. Nine of the ten local yeast strains were selected for their good oenological properties and their suitability as a wine starter culture.

Population Dynamics ofSaccharomyces cerevisiaeduring Spontaneous Fermentation at a British Columbia Winery

Spontaneous fermentation is thought to provide conditions where diversity of indigenous Saccharomyces cerevisiae strains can flourish. A broad diversity of strains may provide unique characteristics to a wine and enhance its sensorial attributes. Population dynamics of S. cerevisiae strains were followed during a spontaneous fermentation of Vitis vinifera L. var. Pinot noir grapes at a winery in Kelowna, British Columbia, Canada, to determine the relative contribution of commercial active dry yeast (ADY) and indigenous strains during fermentation. Grapes were harvested, processed, and fermented separately. Yeasts from grapes were isolated and plated onto culture media. Fermentation tanks were sampled at early, mid, and late stages of fermentation. Yeast strains were isolated onto culture media at each stage. DNA was extracted from yeast cultures and amplified using primers defining six different microsatellite loci. Results were compared with a database comprising DNA fingerprints from commercially produced ADY strains used previously by the winery. Strains isolated from fermentation tanks were all ADY strains closely related or identical to those in the winery database. Strain richness tended to be greater in the mid and final stages than in the early stage. Lalvin ICV D254/Fermol Premier Cru and Red Star Premier Cuvee were the dominant yeasts during the mid and final stages. The former was the only strain found in all tanks in the mid and final stages. An ADY strain, Fermol Arome-Plus, was isolated from both grapes and spontaneous fermentation. This strain could represent a resident strain, where it persists year after year and cycles annually between the winery and vineyard.

Very early acetaldehyde production by industrial Saccharomyces cerevisiae strains: a new intrinsic character

During a general survey of the acetaldehyde-producing properties of commercially available wine yeast strains, we discovered that, although final acetaldehyde production cannot be used as a discriminating factor between yeast strains, initial specific acetaldehyde production rates were of highly interest for classifying yeast strains. This parameter is very closely related to the growth- and fermentation-lag phase durations. We also found that this acetaldehyde early production occurs with very different extent between commercial active dry yeast strains during the rehydration phase and could partially explain the known variable resistance of yeast strains to sulfites. Acetaldehyde production appeared, therefore, as very precocious, strain-dependent, and biomass-independent character. These various findings suggest that this new intrinsic characteristic of industrial fermenting yeast may be likely considered as an early marker of the general fermenting activity of industrial fermenting yeasts. This phenomenon could be particularly important for understanding the ecology of colonization of complex fermentation media by Saccharomyces cerevisiae.

Interactions between Brettanomyces bruxellensis and other yeast species during the initial stages of winemaking

Wine is the product of complex interactions between yeasts and bacteria in grape must. Amongst yeast populations, two groups can be distinguished. The first, named non-Saccharomyces (NS), colonizes, with many other micro-organisms, the surface of grape berries. In the past, NS yeasts were primarily considered as spoilage micro-organisms. However, recent studies have established a positive contribution of certain NS yeasts to wine quality. Amongst the group of NS yeasts, Brettanomyces bruxellensis, which is not prevalent on wine grapes, plays an important part in the evolution of wine aroma. Some of their secondary metabolites, namely volatile phenols, are responsible for wine spoilage. The other group contributing to wine aroma, which is also the main agent of alcoholic fermentation (AF), is composed of Saccharomyces species. The fermenting must is a complex microbial ecosystem where numerous yeast strains grow and die according to their adaptation to the medium. Yeast-yeast interactions occur during winemaking right from the onset of AF. The aim of this study was to describe the interactions between B. bruxellensis, other NS and Saccharomyces cerevisiae during laboratory and practical scale winemaking. Molecular methods such as internal transcribed spacer-restriction fragment length polymorphism and polymerase chain reaction and denaturing gradient gel electrophoresis were used in laboratory scale experiments and cellar observations. The influence of different oenological practices, like the level of sulphiting at harvest time, cold maceration preceding AF, addition of commercial active dry yeasts on B. bruxellensis and other yeast interactions and their evolution during the initial stages of winemaking have been studied. Brettanomyces bruxellensis was the most adapted NS yeast at the beginning of AF, and towards the end of AF it appeared to be more resistant than S. cerevisiae to the conditions of increased alcohol and sugar limitation. Among all NS yeast species, B. bruxellensis is better adapted than other wild yeasts to resist in must and during AF. Moreover, B. bruxellensis appeared to be more tolerant to ethanol stress than S. cerevisiae and after AF B. bruxellensis was the main yeast species in wine. Brettanomyces bruxellensis interacts with other yeast species and adapts to the wine medium as the dominant yeast species at the end of AF. Contamination of B. bruxellensis might take place at the beginning of malolactic fermentation, which is a critical stage in winemaking.