IVAS Session 3 – Posters

Saccharomyces cerevisiae, as the workhorse of alcoholic fermentation, is a major actor of winemaking. In this context, this yeast species uses alcoholic fermentation to convert sugars from the grape must into ethanol and CO2 with an outstanding efficiency: it reaches on average 92% of the maximum theoretical yield of conversion. Moreover, S. cerevisiae is also known for its great genetic diversity and plasticity that is directly related to its living environment, natural or technological and therefore to domestication. This leads to a great phenotypic diversity of metabolites production.

Sterols are a fraction of the eukaryotic lipidome that is essential for the maintenance of the cell membrane integrity and their good functionality. During alcoholic fermentation, they ensure yeast growth, metabolism and viability, as well as resistance to osmotic stress and ethanol inhibition. Two sterol sources can support yeasts to adapt to fermentation stress conditions: ergosterol, produced by yeast in aerobic conditions, and phytosterols, plant sterols found in grape musts imported by yeasts in anaerobiosis. Little is known about the physiological impact of the assimilation of phytosterols in comparison to ergosterol and the influence of sterol type on fermentation kinetics parameters.

SO2 is an additive widely used as antimicrobial in winemaking industry. However, this compound can negatively affect health, so the search for alternatives is currently a line of research of great interest. One of the proposed alternatives to SO2 as an antimicrobial is the use of bioprotection yeasts, which colonize the medium preventing the proliferation of undesirable microorganisms.

Nitrogen and temperature are two important factors that influence wine fermentation and volatile compounds production. Among the different compounds present in the must, nitrogen is an essential nutrient for the management of the fermentation kinetics but it also plays an important role in the synthesis of fermentative aromas. To address the problems related to nitrogen deficiencies, nitrogen additions during alcoholic fermentation have been developed.

Carbonic Maceration (CM) winemaking is typically used in different European regions. But It is paradoxical that being a traditional processing system and widely used in many wineries, some of the phenomena that take place and the parameters that characterize them are barely known. In this vinification system the intact grape clusters are placed in a carbon dioxide (CO2) enriched medium, and they immediately change from a respiratory metabolism to an anaerobic fermentative metabolism called intracellular fermentation, which is carried out by grape enzymes. But some grapes located in the lower zone of the tank are crushed by the weight of the ones above and release must, which is fermented by yeasts.

Botrytis cinerea causes grey mold that results in severe problems for wine makers worldwide. Infected grapes lead to quality deterioration including formation of off-flavors or browning. The latter is caused by the enzyme laccase which is capable of oxidizing a wide range of phenolic compounds. Since the use of conventional pesticides is associated with many concerns of consumers and authorities regarding environmental and health related issues and may result in fungicide resistance, the development of green alternatives is gaining more attention.

Different yeasts were isolated from spontaneous fermentation of Assyrtiko grape must in Santorini Island, Greece. Molecular typing revealed the presence of three Saccharomyces cerevisiae strains (S9, S13, S24) and one strain of the yeast species Nakazawaea ishiwadae (N.i). The four isolated strains were further tested in laboratory scale fermentations of Assyrtiko must in pure inoculation cultures and in sequential inoculation (72 hours) of each S. cerevisiae strain with the strain of N. ishiwadae. All fermentation trials were realised in duplicate.

Varietal thiols are considered for years as key aroma compounds in many wines. Their main origin is the cleavage during alcoholic fermentation of S-conjugate precursors present in grapes and musts, even if the levels of precursors already identified struggle to completely explain the levels of thiols found in wine.

Many microbial starters for the alcoholic and malolactic fermentation processes are commercially available, indicated for diverse wine styles and quality goals. The screening protocols cover a wide range of oenologically relevant features, although some characteristics could also be studied using underexplored powerful techniques. In this study, we applied Fourier Transform Infrared (FTIR) microspectroscopy [1,2] to compare the cell wall biochemical composition and monitor the autolytic process in several wine strains of Saccharomyces cerevisiae.

Microbiomes have crucial roles in maintaining life on Earth, and their functions drive human, animal, plant and environmental health. The microbiome research landscape is developing rapidly and is performed in many different science fields using similar concepts but mostly one (eco)system at-a-time. Thus, we are only starting to unravel and understand the interconnectedness of microbiomes across the (eco)systems.

Ochratoxin A (OTA) is a mycotoxin produced by several filamentous fungi infecting grape bunches (Penicillium and Aspergillus spp.), this toxin pass to must when grapes are crushed and later it is found in wine. Following the evaluations of the toxicity of OTA, European Commission Regulations have been promulgated introducing upper limits for OTA concentrations in various commodities (cereals, cereal products, dried vine fruit, coffee, wine, grape juice, baby foods and dietary foods for special medical purposes).

The addition of fungal chitosan in wine is allowed since 2009 to release some spoilage microorganisms such as Brettanomyces bruxellensis (OIV/OENO 338A/2009; EC 53/2011). This yeast is able to produce volatil phenols and is responsible of organoleptic deviations compromising quality and typicality of red wines [1]. Despite the fact that fungal chitosan is highly renewable, no toxic and non-allergenic, its use remains marginal because this treatment is relatively recent (compare to sulphites treatment) and information are contradictory between different studies described in literature. For all these reasons,

Malolactic fermentation (MLF) is a secondary wine fermentation conducted by lactic acid bacteria (LAB). This fermentation is important in winemaking as it deacidifies the wine, converting L-malic acid into L-lactic acid and carbon dioxide, and it contributes to microbial stability. Wine pH is highly selective, and at pH below 3.5 generally only strains of O. oeni can survive and express malolactic activity, while under more favorable growth conditions above pH 3.5, species of Lactobacillus and Pediococcus may conduct the MLF. Among the LAB species Lactiplantibacillus plantarum strains have shown most interesting results under hot climate conditions, not only for their capacity to induce MLF, but also for their homo-fermentative properties towards hexose sugars, which makes them suitable for induction of MLF in high pH and high alcohol wines, when inoculated at the beginning of alcoholic fermentation.

Certain yeast species belonging to the Pichia genus are known to form a distinctive film on grape must and wine. In a mixed-culture type fermentation, Pichia spp. (P. kluyveri in particular) are known to impart beneficial oenological attributes. In this study, we report on an easy isolation method of Pichia spp. from grape must by exploiting their film-forming capacity on media containing 10% ethanol. We isolated and identified two Pichia species, namely Pichia kudriavzevii and Pichia kluyveri, and subsequently co-inoculated them with Saccharomyces cerevisiae to ferment Gewürztraminer musts. Noteworthy differences included a significant increase in the 2-phenethyl acetate levels with the P. kluyveri co-fermentation and a general increase in ethyl esters with the P. kudriavzevii co-fermentation. Both Pichia co-inoculations yielded higher levels of glycerol in the final wines. Based on all the wine parameters we tested, the P. kluyveri strain that was isolated performed similarly to a commercial P. kluyveri strain.

Infection of the grapes with Botrytis cinerea has a tremendous impact on the resulting crop yield and quality. Well-known problems that are associated with B. cinerea are specific off-flavors, poor filterability, and brownish color in white wines. The development of a B. cinerea infection strongly depends on weather conditions and is highly variable through different vintages. Typical control measures include defoliation and the use of fungicides, which involves high personnel and material costs. They also involve a great risk, especially since the effectiveness and time point of these treatments are difficult to predict.