Aureobasidium pullulans is a yeast-like fungus found on the surface of the grape berries that has been proven to act as a biocontrol agent for the management of grey mould disease caused by Botrytis cinerea. In this work, an indigenous strain of A. pullulans isolated from grape berries and selected according to the in vitro activity against B. cinerea, was used in vineyards of the winery where it originated, in comparison with a commercial product containing two A. pullulans strains with the aim of assessing its effectiveness as a biocontrol agent. The experimental design included daily meteorological data registration and the early defoliation of grapevines as treatments. The monitoring of A. pullulans strains on grape berries by plate counts and molecular methods as well as of B. cinerea symptoms on grape bunches was performed in the different trials from the end of flowering to the harvest time. Results highlighted that although no significant differences (p < 0.05) in the occurrence of B. cinerea were detected according to different treatments, the mean incidence of symptomatic berries ranged from 7 to 16%, with the lowest values recorded in bunches treated with the indigenous A. pullulans strain. The efficacy of the biocontrol agent was affected more by meteorological conditions than the defoliation practice.

Probiotic milk is a class of fermented milk that possesses health-promoting effects, not only due to the lactic acid bacteria (LAB) presence but potentially also to yeast activity. Hence, the aim of this work was to isolate and select yeasts from spontaneous milk fermentations to be used as inoculum, together with LAB, for manufacturing a potentially probiotic acidic low-alcohol fermented milk. Six yeast species were detected from the spontaneous milk fermentation. A screening of 13 yeast strains and 14 previously isolated LAB strains, based on the resistance to bile salts and to acidic conditions, was carried out. The best performing strains were successively tested for in vitro gastrointestinal tolerance. A strain of Kluyveromyces marxianus and a strain of Lactococcus lactis were selected for the manufacturing of two different fermented milk. The values of the main technological and microbiological parameters (pH, organic acids, ethanol, and microbial concentrations) of the experimental milk were in the range of those reported for this category of products. The evaluation of microorganism survival in fermented milk samples subjected to simulated gastrointestinal conditions highlighted a high resistance of both strains. In conclusion, the selected microbial starter culture enabled the setting up of potential probiotic fermented milk.

The inoculation of grape juice with Saccharomyces cerevisiae strains selected from indigenous yeast populations can be a suitable tool to control alcoholic fermentation, contributing to producing wines with typical flavor and aroma and, hence, the demand for native starter cultures is increasing. However, since low amounts of indigenous yeast biomasses are usually required for local winemaking, the industrial production of these yeasts can be expensive. Therefore, in this study, after selecting an indigenous S. cerevisiae strain based on relevant oenological and technological features, a pilot-plant for easy and rapid production of fresh yeast biomass directly in a winery located in Tuscany, was exploited. The selected yeast strain was used as a starter to carry out 25 and 100 hL fermentations and its enological performance was compared with that of the commercial starter normally used in the winery. Chemical and sensory analysis of the resulting wines showed that they differentiated according to the used yeast strain, with the wines produced by the indigenous S. cerevisiae strain being characterized by a distinctive aromatic and sensory profile. In conclusion, the pilot-plant effectively resulted in producing fresh yeast starter cultures in the winery to be successfully used to carry out alcoholic fermentations.

In extra virgin olive oil (EVOO) extraction process, the occurrence of yeasts that could affect the quality of olive oil was demonstrated. Therefore, in this work, at first, the yeasts occurring during different extractive processes carried out in a Tuscany oil mill, at the beginning, in the middle, and the end of the harvesting in the same crop season, were quantified. Then, possible effects on quality of EVOO caused by the predominant yeast species, possessing specific enzymatic activities, were evaluated. Yeast concentrations were higher in extraction processes at the end of the harvesting. Twelve yeast species showing different isolation frequencies during olive oil extractive process and according to the harvesting date were identified by molecular methods. The yeast species dominating olive oil samples from decanter displayed enzymatic activities, potentially affecting EVOO quality according to zymogram analysis. HS-SPME-GC-MS analysis of the volatile compounds in commercial EVOO, inoculated with three yeast species (Nakazawaea molendini-olei, Nakazawaea wickerhamii, Yamadazyma terventina), pointed out significant differences depending on the strain inoculated. In conclusion, during the olive oil extractive processes, some yeast species colonize the extraction plant and may influence the chemical and sensory characteristics of EVOO depending on the cell concentrations and their enzymatic capabilities.

Brettanomyces bruxellensis is found in several fermented matrices and produces relevant alterations to the wine quality. The methods usually used to identify B. bruxellensis contamination are based on conventional microbiological techniques that require long procedures (15 days), causing the yeast to spread in the meantime. Recently, a flow cytometry kit for the rapid detection (1–2 h) of B. bruxellensis in wine has been developed. The feasibility of the method was assessed in a synthetic medium as well as in wine samples by detecting B. bruxellensis in the presence of other yeast species (Saccharomyces cerevisiae and Pichia spp.) and at the concentrations that produce natural contaminations (up to 10⁵ cells/mL), as well as at lower concentrations (10³–10² cells/mL). Wine samples naturally contaminated by B. bruxellensis or inoculated with four different strains of B. bruxellensis species together with Saccharomyces cerevisiae and Pichia spp., were analyzed by flow cytometry. Plate counts were carried out in parallel to flow cytometry. We provide evidence that flow cytometry allows the rapid detection of B. bruxellensis in simple and complex mixtures. Therefore, this technique has great potential for the detection of B. bruxellensis and could allow preventive actions to reduce wine spoilage.