The Riboflavin Metabolism in Four Saccharomyces cerevisiae Wine Strains: Assessment in Oenological Condition and Potential Implications with the Light-Struck Taste.

Riboflavin (RF), or vitamin B2, is an essential compound for yeast growth and a precursor of the flavin coenzymes, flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD), involved in redox and non-redox processes. RF is a photosensitive compound involved in the light-struck taste (LST), a fault causing the formation of off-flavors that can develop when the wine is exposed to light in the presence of methionine (Met), as well. As both RF and Met can be associated with detrimental changes in wines, a better comprehension of its yeast-mediated production is relevant to predict the maintenance of the desired character of the wine. This study aims at assessing the production of flavin derivatives (FDs) and Met by S. cerevisiae oenological starters under laboratory conditions. The results showed the presence of extra- and intracellular FDs, and Met is a strain-dependent characteristic being also affected by the initial content of RF in the medium. This finding was confirmed when the winemaking was carried out in a relevant environment. Our results evidenced the important impact of the yeast strain on the content of RF and its derivatives.

Identifying the Main Drivers in Microbial Diversity for Cabernet Sauvignon Cultivars from Europe to South Africa: Evidence for a Cultivar-Specific Microbial Fingerprint.

Microbial diversity in vineyards and in grapes has generated significant scientific interest. From a biotechnological perspective, vineyard and grape biodiversity has been shown to impact soil, vine, and grape health and to determine the fermentation microbiome and the final character of wine. Thus, an understanding of the drivers that are responsible for the differences in vineyard and grape microbiota is required. The impact of soil and climate, as well as of viticultural practices in geographically delimited areas, have been reported. However, the limited scale makes the identification of generally applicable drivers of microbial biodiversity and of specific microbial fingerprints challenging. The comparison and meta-analysis of different datasets is furthermore complicated by differences in sampling and in methodology. Here we present data from a wide-ranging coordinated approach, using standardized sampling and data generation and analysis, involving four countries with different climates and viticultural traditions. The data confirm the existence of a grape core microbial consortium, but also provide evidence for country-specific microbiota and suggest the existence of a cultivar-specific microbial fingerprint for Cabernet Sauvignon grape. This study puts in evidence new insight of the grape microbial community in two continents and the importance of both location and cultivar for the definition of the grape microbiome.

Valorisation of Bovine Sweet Whey and Sunflower Press Cake Blend through Controlled Fermentation as Platform for Innovative Food Materials.

The current environmental challenge is pushing food systems towards more sustainable models of production that require reorganizing of processes by re-using side products still containing nutrients. This work aimed at valorising a mix of bovine sweet whey and sunflower press cake, through targeted fermentation. After preliminary screening based on growth rate, final pH, lactose/galactose assimilation, phytase activity, six Lactic Acid Bacteria strains (Lacticaseibacillus casei, L. paracasei (2), Lactococcus lactis, Lentilactobacillus parakefiri and Leuconostoc pseudomesenteroides) and three yeasts (Kluyveromyces lactis, K. marxianus and Torulaspora delbrueckii) were co-cultivated in pairs in microcosms (1-part ground press cake: 4-parts whey). All tested microorganisms were able to grow and acidify the blend: the LAB counts increased during the incubation (26 °C for 48 h) of +2.80 log CFU/g, whereas yeasts counts were of +1.98 log CFU/g, with significant differences among the different associations (p < 0.01). Mould counts were always <3 log CFU/g. Interestingly, the bacterial contaminants count significantly varied in samples with different pairs of strains (p < 0.001). Acidification level, acetic acid and ethanol contents were the limiting factors affecting the growth of spoilage micro-organisms. Best performances were attained in microcosms inoculated with L. lactis or L. paracasei and K. lactis or K. marxianus.

Culturable Yeast Diversity of Grape Berries from Vitis vinifera ssp. sylvestris (Gmelin) Hegi.

Vitis vinifera L. ssp. sylvestris (Gmelin) Hegi is recognized as the dioecious parental generation of today's cultivars. Climatic change and the arrival of pathogens and pests in Europe led it to be included on the International Union for Conservation of Nature (IUCN) Red List of Threatened Species in 1997. The present work focused on the study of culturable yeast occurrence and diversity of grape berries collected from wild vines. Sampling was performed in 29 locations of Azerbaijan, Georgia, Italy, Romania, and Spain. In total, 3431 yeast colonies were isolated and identified as belonging to 49 species, including Saccharomyces cerevisiae, by 26S rDNA D1/D2 domains and ITS region sequencing. Isolates of S. cerevisiae were also analyzed by SSR-PCR obtaining 185 different genotypes. Classical ecology indices were used to obtain the richness (S), the biodiversity (H'), and the dominance (D) of the species studied. This study highlights the biodiversity potential of natural environments that still represent a fascinating source of solutions to common problems in winemaking.

Validation of a Standard Protocol to Assess the Fermentative and Chemical Properties of Saccharomyces cerevisiae Wine Strains.

This paper reports on a common experiment performed by 17 Research Units of the Italian Group of Microbiology of Vine and Wine (GMVV), which belongs to the Scientific Society SIMTREA, with the aim to validate a protocol for the characterization of wine strains of Saccharomyces cerevisiae. For this purpose, two commercial S. cerevisiae strains (EC 1118 and AWRI796) were used to carry out inter-laboratory-scale comparative fermentations using both synthetic medium and grape musts and applying the same protocol to obtain reproducible, replicable, and statistically valid results. Ethanol yield, production of acetic acid, glycerol, higher alcohols, and other volatile compounds were assessed. Moreover, the Fourier transform infrared spectroscopy was also applied to define the metabolomic fingerprint of yeast cells from each experimental trial. Data were standardized as unit of compounds or yield per gram of sugar (glucose and fructose) consumed throughout fermentation, and analyzed through parametric and non-parametric tests, and multivariate approaches (cluster analysis, two-way joining, and principal component analysis). The results of experiments carried out by using synthetic must showed that it was possible to gain comparable results from three different laboratories by using the same strains. Then, the use of the standardized protocol on different grape musts allowed pointing out the goodness and the reproducibility of the method; it showed the main traits of the two yeast strains and allowed reducing variability amongst independent batches (biological replicates) to acceptable levels. In conclusion, the findings of this collaborative study contributed to the validation of a protocol in a specific synthetic medium and in grape must and showed how data should be treated to gain reproducible and robust results, which could allow direct comparison of the experimental data obtained during the characterization of wine yeasts carried out by different research laboratories.

Is the Consumer Ready for Innovative Fruit Wines? Perception and Acceptability of Young Consumers.

The consumption of traditional wine has decreased in Europe during the last fifteen years. In parallel, new wine alternatives obtained by blending wines and fruit juices or by flavoring wines with artificial or natural flavors have appeared on the market. Recently, an innovative fruit wine obtained by co-fermentation of grape must and kiwi juice has been proposed and its potential of attraction for consumers should be exploited. To assess the potential consumer acceptability and expectations towards this new product, an online choice experiment has been conducted involving a consumer group of young adults (18-35 years old; n = 373). After the data collection, participants were divided into two groups according to whether they had already tasted a fruit wine (neophiles) or had never tasted it (new entries). For each group, the individual's responses (on wine consumption habits, expectations and willingness to consume and pay a fruit wine) were analyzed through Principal Component Analysis. Different consumption styles and expectation patterns were defined in the two groups. However, in general, neophiles showed consumption patterns based on the evaluation of fruit quality, sales format, alcoholic content and the presence or not of bubbles, not giving importance to the brand. In contrast, new entries' responses identified consumption patterns driven by the willingness to pay for a new product, the product value for money and packaging features. Differences between the two groups in expectations about the product sensory characteristics also emerged. These findings should contribute to this area of study by integrating environmental, economic and social dimensions and addressing food innovation and sustainability in the fruit and wine chains.

The Role of Yeasts as Biocontrol Agents for Pathogenic Fungi on Postharvest Grapes: A Review.

In view of the growing concern about the impact of synthetic fungicides on human health and the environment, several government bodies have decided to ban them. As a result, a great number of studies have been carried out in recent decades with the aim of finding a biological alternative to inhibit the growth of fungal pathogens. In order to avoid the large losses of fruit and vegetables that these pathogens cause every year, the biological alternative's efficacy should be the same as that of a chemical pesticide. In this review, the main studies discussed concern Saccharomyces and non-Saccharomyces yeasts as potential antagonists against phytopathogenic fungi of the genera Penicillium and Aspergillus and the species Botrytis cinerea on table grapes, wine grapes, and raisins.

Investigating the growth kinetics in sourdough microbial associations.

In this study we investigated the effect of the single strain in stabilization of type I sourdough microbial associations by crossing six different Fructilactobacillus sanfranciscensis with five Kazachstania humilis strains. Furthermore, we compared three predictive models, Zwitwering based on Gompertz's equation, Baranyi and Roberts' function and Schiraldi's function to evaluate which one best fitted the experimental data in determining the behaviour of co-cultivated microorganisms. Specific growth rates (µm) and lag time (λ) values for each mixed population were assessed. Results showed that the different F. sanfranciscensis strains significantly steer the growth kinetics within the pair and affect the ratio bacterial/yeast cells, as data analysis confirmed, whereas K. humilis accommodates to the bacterial strain. To compare the growth models, Root Mean Square (RMS) values were calculated for each predicted curve by implementing an algorithm based on an iterative process to minimize the deviation among observed and calculated data. Schiraldi's function performed better than the others, revealing, on average, the smallest RMS values and providing the best fitting for over 70% of co-cultivation experiments. Models prove to be consistent in predicting growth kinetics of microbial consortia too.

Tryptophan Derivatives by Saccharomyces cerevisiae EC1118: Evaluation, Optimization, and Production in a Soybean-Based Medium.

Given the pharmacological properti es and the potential role of kynurenic acid (KYNA) in human physiology and the pleiotropic activity of the neurohormone melatonin (MEL) involved in physiological and immunological functions and as regulator of antioxidant enzymes, this study aimed at evaluating the capability of Saccharomyces cerevisiae EC1118 to release tryptophan derivatives (dTRPs) from the kynurenine (KYN) and melatonin pathways. The setting up of the spectroscopic and chromatographic conditions for the quantification of the dTRPs in LC-MS/MS system, the optimization of dTRPs' production in fermentative and whole-cell biotransformation approaches and the production of dTRPs in a soybean-based cultural medium naturally enriched in tryptophan, as a case of study, were included in the experimental plan. Variable amounts of dTRPs, with a prevalence of metabolites of the KYN pathway, were detected. The LC-MS/MS analysis showed that the compound synthesized at highest concentration is KYNA that reached 9.146 ± 0.585 mg/L in fermentation trials in a chemically defined medium at 400 mg/L TRP. Further experiments in a soybean-based medium confirm KYNA as the main dTRPs, whereas the other dTRPs reached very lower concentrations. While detectable quantities of melatonin were never observed, two MEL isomers were successfully measured in laboratory media.

Transcriptomics unravels the adaptive molecular mechanisms of Brettanomyces bruxellensis under SO2 stress in wine condition.

Sulfur dioxide is generally used as an antimicrobial in wine to counteract the activity of spoilage yeasts, including Brettanomyces bruxellensis. However, this chemical does not exert the same effectiveness on different B. bruxellensis yeasts since some strains can proliferate in the final product leading to a negative sensory profile due to 4-ethylguaiacol and 4-ethylphenol. Thus, the capability of deciphering the general molecular mechanisms characterizing this yeast species' response in presence of SO2 stress could be considered strategic for a better management of SO2 in winemaking. A RNA-Seq approach was used to investigate the gene expression of two strains of B. bruxellensis, AWRI 1499 and CBS 2499 having different genetic backgrounds, when exposed to a SO2 pulse. Results revealed that sulphites affected yeast culturability and metabolism, but not volatile phenol production suggesting that a phenotypical heterogeneity could be involved for the SO2 cell adaptation. The transcriptomics variation in response to SO2 stress confirmed the strain-related response in B. bruxellensis and the GO analysis of common differentially expressed genes showed that the detoxification process carried out by SSU1 gene can be considered as the principal specific adaptive response to counteract the SO2 presence. However, nonspecific mechanisms can be exploited by cells to assist the SO2 tolerance; namely, the metabolisms related to sugar alcohol (polyols) and oxidative stress, and structural compounds.

LC-MS/MS-Based Profiling of Tryptophan-Related Metabolites in Healthy Plant Foods.

Food plants contain hundreds of bioactive phytochemicals arising from different secondary metabolic pathways. Among these, the metabolic route of the amino acid Tryptophan yields a large number of plant natural products with chemically and pharmacologically diverse properties. We propose the identifier "indolome" to collect all metabolites in the Tryptophan pathway. In addition, Tryptophan-rich plant sources can be used as substrates for the fermentation by yeast strains to produce pharmacologically active metabolites, such as Melatonin. To pursue this technological development, we have developed a UHPLC-MS/MS method to monitor 14 Tryptophan, Tryptamine, amino-benzoic, and pyridine metabolites. In addition, different extraction procedures to improve the recovery of Tryptophan and its derivatives from the vegetal matrix were tested. We investigated soybeans, pumpkin seeds, sesame seeds, and spirulina because of their botanical diversity and documented healthy effects. Four different extractions with different solvents and temperatures were tested, and water extraction at room temperature was chosen as the most suitable procedure to extract the whole Tryptophan metabolites pattern (called by us "indolome") in terms of ease, high efficiency, short time, low cost, and sustainability. In all plant matrices, Tryptophan was the most abundant indole compound, while the pattern of its metabolites was different in the diverse plants extracts. Overall, 5-OH Tryptamine and Kynurenine were the most abundant compounds, despite being 100-1000-fold lower than Tryptophan. Melatonin was undetected in all extracts, but sesame showed the presence of a Melatonin isomer. The results of this study highlight the variability in the occurrence of indole compounds among diverse food plants. The knowledge of Tryptophan metabolism in plants represents a relevant issue for human health and nutrition.

Production of melatonin and other tryptophan derivatives by Oenococcus oeni under winery and laboratory scale.

Malolactic fermentation (MLF) in Valtellina Superiore DOCG red wine was monitored in 4 cellars and the final products were analysed to determine the content of melatonin (MEL) and other tryptophan (TRP) derivatives, including tryptophan ethyl ester (TEE) and MEL isomers (MISs), and to isolate predominant O. oeni strains. MEL and TEE significantly increased in wines after MLF from two cellars out of four. Six strains were isolated during the MLF of red wines and under laboratory scale, in rich and synthetic wine cultural media, together with other four O. oeni strains able to trigger the MLF. Results showed that the presence of stressful growth factors, like ethanol and acid pH, has a pivotal role in triggering the release of TEE by oenococci. Indeed, all the strains became capable to produce also MEL and MISs, together with TEE. under harsh growth conditions, as in a synthetic wine medium. The production of these compounds was strain-dependent and a maximum amount of 0.0078 ±â€¯0.0023 ngT/mL (UMB472) and 619.85 ±â€¯196.16 ngT/mL (UMB436) of MEL and TEE was obtained, respectively. In particular, different MISs were detected under oenological and laboratory scale suggesting that other factors (i.e. technological and/or physico-chemical) could affect the synthesis of TRP derivatives.

Set-Up of Bacterial Cellulose Production From the Genus Komagataeibacter and Its Use in a Gluten-Free Bakery Product as a Case Study.

The use of bacterial cellulose (BC) in food systems is still limited due to production costs. Nine clones belonging to Komagataeibacter hansenii, Komagataeibacter nataicola, Komagataeibacter rhaeticus, Komagataeibacter swingsii, and Komagataeibacter xylinus species were screened for cellulose productivity in growth tests with five different carbon sources and three nitrogen sources. The water-holding and rehydration capacities of the purified cellulose were determined. The structure of the polymer was investigated through nuclear magnetic resonance (NMR) spectroscopy, attenuated total reflection Fourier transform infrared (ATR-FT-IR) spectroscopy and X-ray diffraction (XRD) analysis, and observed by scanning electron microscope (SEM). Natural mutants of K. rhaeticus LMG 22126T and K. swingsii LMG 22125T showed different productivity. The factors "bacterial isolate" and "nitrogen source" significantly affected the production of cellulose (p < 0.01) rather than the factor "carbon source" (p = 0.15). However, on average, the best conditions for increasing yield were found in medium containing glucose and peptone. Water-holding capacity (WHC) values ranged from 10.7 to 42.3 (g water/g cellulose) with significant differences among strains (p < 0.01), while the rehydration capacity varied from 4.2 to 9.3 (g water/g cellulose). A high crystallinity (64-80%) was detected in all samples with Iα fractions corresponding to 67-93%. The ATR-FT-IR spectra and the XRD patterns confirmed the expected structure. BC made by GVP isolate of K. rhaeticus LMG 22126T, which was the strain with the highest yield, was added to a gluten-free bread formulation. Results obtained from measurements of technological parameters in dough leavening and baking trials were promising for implementation in potential novel foods.

Corrigendum: Investigating the Effect of Selected Non-Saccharomyces Species on Wine Ecosystem Function and Major Volatiles.

[This corrects the article DOI: 10.3389/fbioe.2018.00169.].

Innovative Alcoholic Drinks Obtained by Co-Fermenting Grape Must and Fruit Juice.

In this study, Cabernet Sauvignon and Chardonnay musts, and fruit juices from cherry, kiwi, peach, and strawberry were co-fermented with Saccharomyces cerevisiae EC1118 and Torulaspora delbrueckii UMY196 at two different proportions (80:20 (v/v) and 60:40 (v/v)). The most pleasant fruit-based drink was obtained with Cabernet Sauvignon must and kiwi juice in a proportion of 60:40 and fermented with T. delbrueckii. This beverage was produced in higher volume to simulate a scale-up, and the aromatic profile, sensory description, and consumer acceptability were determined. The most powerful odorants of the kiwi-based drink were ethyl octanoate, phenylethanal, ethyl hexanoate, vinyl-guaiacol, benzaldehyde, and nonanal, for which the odor activity values were 21.1, 3.3, 2.6, 2.2, 1.9, and 1.6, respectively. These findings were in accordance with the sensory analysis, since the emerged descriptors were fruity (ethyl octanoate), honey and floral (phenylethanal), apple and peach (ethyl hexanoate), and citrus (nonanal). The consumers judged the kiwi-based drink acceptable (67%) and 39% of them would buy it. The reliable fermentation of a grape must/fruit juice was demonstrated. The kiwi-based drink represents an innovative and pleasant beverage with a positive impact on sustainability as its production can limit the loss of fresh fruits, as well as contribute to the enological field.

Assessment of Tryptophan, Tryptophan Ethylester, and Melatonin Derivatives in Red Wine by SPE-HPLC-FL and SPE-HPLC-MS Methods.

Melatonin (MEL) is an indoleamine produced mainly by the pineal gland in vertebrates. It plays a significant role in the regulation of circadian rhythms, mitigation of sleeping disorders, and jet lag. This compound is synthetized from tryptophan (TRP) and it has been found in seeds, fruits, and fermented beverages, including wine. Wine is also a source of other tryptophan derivatives, the tryptophan ethylester (TEE) and MEL isomers (MISs), for which the biological properties need to be elucidated. An analytical method for the simultaneous quantification of TRP, TEE, and MEL was developed by a Solid Phase Extraction (SPE) of a preconcentration of wine followed by high performance liquid chromatography (HPLC) analysis either with fluorescence or mass spectrometer detectors. The analytical method showed a relative standard deviation (RSD) lower than 8%, except for TRP (RSD 10.5% in wine). The recovery was higher than 76%. The versatility of SPE preconcentrations allowed for the adequate preconcentration of wine sample as well as detection of low concentrations, an important aspect especially for MEL (detection limit 0.0023 µg/L). The proposed method proved to be suitable for assessing the investigated compounds in some red wine samples, where 74.4⁻256.2 µg/L and 0.038⁻0.063 µg/L of TEE and MEL were detected, respectively. Five MISs were also found in wine samples in concentrations up to 1.97 µg/L.

Assessment of Microbial Populations in the Manufacture of Vacuum-Packaged Ready-to-Eat Roast Beef and in a Related Production Plant.

Some microbiological criteria were monitored for 6 months in vacuum-packaged roast beef (15 production batches), raw beef (10 batches), and other meat products (12 batches) produced in an Italian small to medium-size enterprise. Fifty-five environmental swab samples also were analyzed. The main bacterial groups were identified by cultural methods according to International Organization for Standardization standards. Listeria monocytogenes was enumerated with the most-probable-number protocol, and species identification was confirmed with a specific PCR assay. Immediately after vacuum packaging, all ready-to-eat (RTE) products had low mean aerobic colony counts (<102 to 2.4 × 102 CFU g-1), anaerobic colony counts (1.6 to 6.5 × 101 CFU g-1), Enterobacteriaceae counts (1.1 to 1.4 × 101 CFU g-1), and Escherichia coli counts (generally below the detection limit). Nevertheless, the prevalence of L. monocytogenes in these samples was 3.7%. In roast beef samples, the aerobic and anaerobic colony counts reached unacceptable levels (>106 CFU g-1) after 14 days of refrigerated storage. Because the prevalence of L. monocytogenes increased to 13.3% during storage, a substantial reduction in the shelf life of these products is recommended. Surfaces without direct contact with food (floors and drains) had the highest mean counts for aerobic colonies (8.0 × 103 to 9.5 × 105 CFU/cm2), anaerobic colonies (2.9 × 103 to 3.2 × 104 CFU/cm2), Enterobacteriaceae (1.5 × 101 to 8.4 × 101 CFU/cm2), and E. coli (6.0 to 7.7 CFU/cm2). The levels of L. monocytogenes on direct food contact surfaces were below the detection limit, but more than 25% of floor samples were contaminated. These results reveal the persistence of L. monocytogenes in food processing environments, although at very low levels, posing a high risk of postcooking recontamination for RTE products. To improve hygienic conditions and reduce cross-contamination, an increase in operator awareness and a reassessment of surface sanitization protocols are needed.

Investigating the Effect of Selected Non-Saccharomyces Species on Wine Ecosystem Function and Major Volatiles.

Natural alcoholic fermentation is initiated by a diverse population of several non-Saccharomyces yeast species. However, most of the species progressively die off, leaving only a few strongly fermentative species, mainly Saccharomyces cerevisiae. The relative performance of each yeast species is dependent on its fermentation capacity, initial cell density, ecological interactions as well as tolerance to environmental factors. However, the fundamental rules underlying the working of the wine ecosystem are not fully understood. Here we use variation in cell density as a tool to evaluate the impact of individual non-Saccharomyces wine yeast species on fermentation kinetics and population dynamics of a multi-species yeast consortium in synthetic grape juice fermentation. Furthermore, the impact of individual species on aromatic properties of wine was investigated, using Gas Chromatography-Flame Ionization Detector. Fermentation kinetics was affected by the inoculation treatment. The results show that some non-Saccharomyces species support or inhibit the growth of other non-Saccharomyces species in the multi-species consortium. Overall, the fermentation inoculated with a high cell density of Starmerella bacillaris displayed the fastest fermentation kinetics while fermentation inoculated with Hanseniaspora vineae showed the slowest kinetics. The production of major volatiles was strongly affected by the treatments, and the aromatic signature could in some cases be linked to specific non-Saccharomyces species. In particular, Wickerhamomyces anomalus at high cell density contributed to elevated levels of 2-Phenylethan-1-ol whereas Starm. bacillaris at high cell density resulted in the high production of 2-methylpropanoic acid and 3-Hydroxybutanone. The data revealed possible direct and indirect influences of individual non-Saccharomyces species within a complex consortium, on wine chemical composition.

Wild Grape-Associated Yeasts as Promising Biocontrol Agents against Vitis vinifera Fungal Pathogens.

The increasing level of hazardous residues in the environment and food chains has led the European Union to restrict the use of chemical fungicides. Thus, exploiting new natural antagonistic microorganisms against fungal diseases could serve the agricultural production to reduce pre- and post-harvest losses, to boost safer practices for workers and to protect the consumers' health. The main aim of this work was to evaluate the antagonistic potential of epiphytic yeasts against Botrytis cinerea, Aspergillus carbonarius, and Penicillium expansum pathogen species. In particular, yeast isolation was carried out from grape berries of Vitis vinifera ssp sylvestris populations, of the Eurasian area, and V. vinifera ssp vinifera cultivars from three different farming systems (organic, biodynamic, and conventional). Strains able to inhibit or slow the growth of pathogens were selected by in vitro and in vivo experiments. The most effective antagonist yeast strains were subsequently assayed for their capability to colonize the grape berries. Finally, possible modes of action, such as nutrients and space competition, iron depletion, cell wall degrading enzymes, diffusible and volatile antimicrobial compounds, and biofilm formation, were investigated as well. Two hundred and thirty-one yeast strains belonging to 26 different species were isolated; 20 of them, ascribed to eight species, showed antagonistic action against all molds. Yeasts isolated from V. vinifera ssp sylvestris were more effective (up to 50%) against B. cinerea rather than those isolated from V. vinifera ssp vinifera. Six strains, all isolated from wild vines, belonging to four species (Meyerozyma guilliermondii, Hanseniaspora uvarum, Hanseniaspora clermontiae, and Pichia kluyveri) revealed one or more phenotypical characteristics associated to the analyzed modes of antagonistic action.