Macrowine 2021
IVES 9 IVES Conference Series 9 Characterization of Brettanomyces bruxellensis biofilm, a resistance strategy to persist in wine-related environments

Characterization of Brettanomyces bruxellensis biofilm, a resistance strategy to persist in wine-related environments

Abstract

AIM: Biofilm is a resistance mechanism deployed by microorganisms to adapt to stresses, leading to their persistence in the environment. In the case of Brettanomyces bruxellensis, a wine spoilage yeast, knowledge about its capacity to form biofilm remains limited although this potential strategy could explain its recurring presence in cellars. In this study, we propose to investigate the capacity of strains of B. bruxellensis to form biofilm according to different conditions and to characterize its structure.

METHODS: Sixty-five isolates of B. bruxellensis were sampled from a cellar and discriminated into genetic groups by microsatellite analysis. The capacity of selected strains to adhere and form biofilm has been investigated in different conditions of media and supports, related to wine environment. The structure of the biofilm and its components were explored using several microscopic techniques as confocal laser scanning microscopy, electronic microscopy and epifluorescence microscopy.

RESULTS: Twelve strains among the 65 isolates were selected and have showed the capacity to form biofilm on polystyrene surface. Microscopic observations of the biofilm revealed microcolonies, filamentous cells and extracellular polymeric substances despite a small thickness. Then, the study of the impact of wine on B. bruxellensis biofilm revealed biofilm cell released and growth of these released biofilm cells, probable contamination source of the wine. Finally, for both planktonic and biofilm lifestyles in wine, a new chlamydospore-like element was observed for B. bruxellensis, described as a resistance form in other fungi.

CONCLUSIONS: This study validates the capacity of B. bruxellensis to form biofilm and highlights structural element of this mode of life. Moreover, an additional resistance strategy was suggested through the description of the chlamydospore-like elements. New insights into the persistence of B. bruxellensis during the winemaking process and in wineries have emerged.

DOI:

Publication date: September 3, 2021

Issue: Macrowine 2021

Type: Article

Authors

Manon Lebleux, Hany ABDO, Louise BASMACIYAN, Julie LAURENT, Chloé ROULLIER-GALL, Hervé ALEXANDRE, Michèle GUILLOUX-BENATIER, Stéphanie WEIDMANN, Sandrine ROUSSEAUX

Univ. Bourgogne Franche-Comté, AgroSup Dijon, PAM UMR A 02.102, VAlMiS Laboratory-IUVV, Dijon, France

Contact the author

Keywords

Brettanomyces bruxellensis, biofilm, wine spoilage

Citation

Related articles…

First step in the preparation of a soil map of the Protected Designation of Origin Valdepeñas (Central, Spain)

This work is a first step to make a map of vineyard soils. The characterization of the soils of the Protected Designation of Origin (D.P.O.) Valdepeñas will allow to group the studied profiles according to their physico-chemical characteristics and the concentrations of most relevant chemical elements. 90 soil profiles were analysed throughout the territory and the soils were sampled and described according to FAO (2006) and classified according to and Soil Taxonomy (2014). All samples were air dried, sieved and some physico-chemical parameters were determined following standard protocols. Also, major and trace elements were analysed by X-ray fluorescence. The statistically study was made using the SPSS program. Trend maps were made using the ArcGIS program. The studied soils have the following average properties: pH, 8.3; electrical conductivity, 0,20 dS/m (low); clay, 18.8% (medium) and CaCO3, 17.1% (high). In the study for the major elements. The major elements of these soils are Si, followed by Ca and Al, with an average content of 203.7 g/kg, 105.5 g/kg and 74.0 g/kg respectively. On the other hand, 27 trace elements have been studied. Of all of them, it can be highlighted the average values of Ba (361.8 mg/kg), Sr (129.3 mg/kg), Rb (83.4 mg/kg), V (74.2 mg/kg) and Ce (70.6 mg/kg). Ba, V and Ce values are higher and the values of Sr and Rb are lower to those found in the literature. The discriminant analysis shows a percentage of grouping of 91%. The content of chemical elements together with the physico-chemical characteristics allows grouping the soils in 4 group according to their order in the classification to Soil Taxonomy; due to the importance of the Calcisols in Castilla-La Mancha, it has been decided to establish them as their own group even if they do not appear in Soil Taxonomy classification.

Evaluation of climate change impacts at the Portuguese Dão terroir over the last decades: observed effects on bioclimatic indices and grapevine phenology

In the last decades the growers of the Portuguese Dão winegrowing region (center of Portugal) are experiencing changes in climate that are influencing either grape phenology berry health and ripening. Aiming to study the relationships between climate indices (CI), seasonal weather and grapevine phenology, in this work long-term climate and phenological data collected at the experimental vineyard of the Portuguese Dão research centre between 1958 and 2019 (61 years) for the red variety Touriga Nacional, was analyzed. The trends over time for the classical temperature-based indices (Growing Season Temperature – GST -, Growing Degree Days – GDD, Huglin Index – HI and Cool Night Index – CI) presented a significantly positive slope while the Dryness Index (DI) showed a negative trend over the last 61 years. Regarding grapevine phenology, an average advance of 4.5 days per decade in the harvest day was observed throughout the last 61 years. Consequently, the weather conditions during the ripening period have changed, showing an increasing trend over time in the average temperature (higher magnitude in the maximum than in the minimum temperature) and a decrease in the accumulated rainfall. A regression analysis showed that ~50% of harvest date variability over years was explained by the temperature-based indices variability. These observed effects of climate change on bioclimatic indices and corresponding anticipation of harvest date can still be considered advantageous for the Dão terroir as it allows to achieve an optimal berry ripening before the common equinox rains and, therefore, avoid the potential negative impacts of the rainfall on berry health and composition.

Variations of soil attributes in vineyards influence their reflectance spectra

Knowledge on the reflectance spectrum of soil is potentially useful since it carries information on soil chemical composition that can be used to the planning of agricultural practices. If compared with analytical methods such as conventional chemical analysis, reflectance measurement provides non-destructive, economic, near real-time data. This paper reports results from reflectance measurements performed by spectroradiometry on soils from two vineyards in south Brazil. The vineyards are close to each other, are on different geological formations, but were subjected to the same management. The objective was to detect spectral differences between the two areas, correlating these differences to variations in their chemical composition, to assess the technique’s potential to predict soil attributes from reflectance data.To that end, soil samples were collected from ten selected vine parcels. Chemical analysis yield data on concentration of twenty-one soil attributes, and spectroradiometry was performed on samples. Chemical differences significant to a 95% confidence level between the two studied areas were found for six soil attributes, and the average reflectance spectra were separated by this same level along most of the observed spectral domain. Correlations between soil reflectance and concentrations of soil attributes were looked for, and for ten soil traits it was possible to define wavelength domains were reflectance and concentrations are correlated to confidence levels from 95% to 99%. Partial Least Squares Regression (PLSR) analyses were performed comparing measured and predicted concentrations, and for fifteen out of 21 soil traits we found Pearson correlation coefficients r > 0.8. These preliminary results, which have to be validated, suggest that variations of concentration in the investigated soil attributes induce differences in reflectance that can be detected by spectroradiometry. Applications of these observations include the assessment of the chemical content of soils by spectroradiometry as a fast, low-cost alternative to chemical analytical methods.

Effect of vigour and number of clusters on eonological parameters and metabolic profile of Cabernet Sauvignon red wines

Vegetative growth and yield are reported to affect grape and wine quality. They can be controlled through different techniques linked to vine management. The objective of this research was to determine the effect of vine vigour and number of clusters per vine on physicochemical composition and phenolic profile of red wines. The experiment was carried out during two vegetative cycles, with cv. Cabernet Sauvignon grafted onto Paulsen 1103. Three vine vigour were defined, according to shoot weight at previous harvests, being low, medium and high. Five treatments of number of clusters were used for each vigour, with 15, 22, 29, 36, and 45 clusters per vine. Grapes from all treatments were harvested in the same day from Brix and total acidity criteria. Thirty days after bottling, classical analyzes and phenolic compounds were performed. As results, different responses were obtained from each vintage. In 2020, a dry season from veraison to harvest, grapes and wines obtained from low vigour treatment and 45 clusters per vine was the highest in sugar and alcohol content respectively, while grapes and wines from high vigour and 15 clusters presented the lowest sugar and alcohol content. Total anthocyanins were higher in treatment with low vigour and 15 clusters, while the lowest amounts were found in low vigour with 45 clusters, as well as medium and high vigour with 36 clusters per vine. Total tannins were higher in high vigour with 22 clusters and medium vigour with 29 clusters, while were lower in low vigour with 36 clusters. In 2021, a wet season at harvest, responses were different, and great variations were observed between treatments. As conclusions, yield and vine vigour had strong influence on grape and wine quality, promoting different enological potentials on which can be indicated/used for aging strategies of red and even rosé wines.

Modulation of berry composition by different vineyard management practices

High concentration of sugars in grapes and alcohol in wines is one of the consequences of climate change on viticulture production in several wine-growing regions. In order to investigate the possibilities of adaptation of vineyard management practices aimed to reduce the accumulation of sugar during the maturation phase without reducing the accumulation of anthocyanins in grapes, a study with severe shoot trimming, shoot thinning, cluster thinning and date of harvest was conducted on Merlot variety in Istria region (Croatia), under the Mediterranean climate. Four factors which may affect grape maturation and its composition at harvest were investigated in a two-years experiment; severe shoot trimming applied at veraison when >80% of berries changed colour (in comparison to untreated control), shoot thinning (0 and 30%), cluster thinning (0 and 30%), and the date of harvest (early and standard harvest dates). Shoot thinning had no significant impact on berry composition, despite the obtained reduction in yield per vine. Lower Brix in grapes were obtained with earlier harvest date and if no cluster thinning was applied, although at the same time a reduction in the concentration of anthocyanins in berries was observed in these treatments. On the other hand, if severe shoot trimming was applied when >80% of berries changed colour, a reduction of Brix was obtained without a negative impact on berry anthocyanins concentration. We conclude that in cases when undesirably high sugar concentrations at harvest are expected, severe shoot trimming at 80% veraison may effectively be used in order to obtain moderate sugar concentration in berries together with the adequate phenolic composition.