Terroir 2004 banner
IVES 9 IVES Conference Series 9 The use of epifluorescence versus plating to monitor the effect of different parameters on microorganisms in wine

The use of epifluorescence versus plating to monitor the effect of different parameters on microorganisms in wine

Abstract

The monitoring of the number of micro-orgranisms in wine is crucial for the wine producer. Traditional counting methods include microscopic enumeration and plating on selective media, which measures the culturability of the cells. The use of epifluorescence microscopy is, however, a method, which can measure both culturability and viability in wine. This method distinguishes between live and dead cells. Research showed that little difference existed between plating and epifluorescence numbers to enumerate lactic acid bacteria in wine. However, a difference exists between these two methods to distinguish between acetic acid bacteria numbers in wine. Plating counting numbers were lower than plate numbers for Acetobacter pasteurianus in wine under anaerobic conditions. This difference was, however, negated by the addition of oxygen to the wine. SO2 additions lowered the culturability of A. pasteurianus at dosages higher than 0.35mg/L molecular SO2, but higher dosages were required to lower epifluorescence intensity, which is an indication of viability. Brettanomyces bruxellensis culturability was inhibited at lower dosages, but total cell numbers according to epifluorescence microscopy were affected at higher molecular SO2 dosages. Epifluorescence microscopy and plating also showed that B. bruxellensis was drastically affected after 120 min after molecular SO2 addition and it’s culturability after only 30 min. An exposure time of 5 min to molecular SO2 reduced the cell’s viability drastically and 45 min completely inhibiting the viability after two days. The bonded form of sulphur dioxide did not affect both micro-organisms. Epifluorescence microscopy can thus be used as a quick alternative to assess micro-organisms numbers and culturability in wine. This technique has both advantages and disadvantages over traditional enumeration methods, which will also be discussed.

DOI:

Publication date: January 12, 2022

Issue: Terroir 2004

Type: Article

Authors

W.J. du Toit (1), I.S. Pretorius (1,2) and A. Lonvaud-Funel (3)

(1) Department of Viticulture and Oenology, Institute for Wine Biotechnology, Stellenbosch University, Victoria Street, ZA 7600 Stellenbosch, South Africa
(2)The Australian Wine Research Institute, Waite Road, Urrbrae, SA 5064 Adealide, Australia
(3) Faculté d’Oenologie, 351, Cours de la Libération, 33405 Talence, France

Contact the author

Tags

IVES Conference Series | Terroir 2004

Citation

Related articles…

Delaying irrigation initiation linearly reduces yield with little impact on maturity in Pinot noir

When to initiate irrigation is a critical annual management decision that has cascading effects on grapevine productivity and wine quality in the context of climate change. A multi-site trial was begun in 2021 to optimize irrigation initiation timing using midday stem water potential (ψstem) thresholds characterized as departures from non-stressed baseline ψstemvalues (Δψstem). Plant material, vine and row spacing, and trellising systems were concomitant among sites, while vine age, soil type, and pruning systems varied. Five target Δψstem thresholds were arranged in an RCBD and replicated eight times at each site: 0.2, 0.4, 0.6, 0.8, and 1.0 MPa (T1, T2, T3, T4, and T5, respectively). When thresholds were reached, plots were irrigated weekly at 70% ETc. Yield components and berry composition were quantified at harvest. To better generalize inferences across sites, data were analyzed by ANOVA using a mixed model including site as a random factor. Across sites, irrigation was initiated at Δψstem = 0.24, 0.50, 0.65, 0.93, and 0.98 MPa for T1, T2, T3, T4, and T5, respectively. Consistent significant negative linear trends were found for several key yield and berry composition variables. Yield decreased by 12.9, 15.9, 19.5, and 27.4% for T2, T3, T4, and T5, respectively, compared to T1 (p < 0.0001) across sites that were driven by similarly linear reductions in berry weight (p < 0.0001). Comparatively, berry composition varied little among treatments. Juice total soluble solids decreased linearly from T1 to T5 – though only ranged 0.9 Brix (p = 0.012). Because producers are paid by the ton, and contracts simply stipulate a target maturity level, first-year results suggest that there is no economic incentive to induce moderate water deficits before irrigation initiation, regardless of vineyard site. Subsequent years will further elucidate the carryover effects of delaying irrigation initiation on productivity over the long term.

Grapevine sugar concentration model in the Douro Superior, Portugal

Increasingly warm and dry climate conditions are challenging the viticulture and winemaking sector. Digital technologies and crop modelling bear the promise to provide practical answers to those challenges. As viticultural activities strongly depend on harvest date, its early prediction is particularly important, since the success of winemaking practices largely depends upon this key event, which should be based on an accurate and advanced plan of the annual cycle. Herein, we demonstrate the creation of modelling tools to assess grape ripeness, through sugar concentration monitoring. The study area, the Portuguese Côa valley wine region, represents an important terroir in the “Douro Superior” subregion. Two varieties (cv. Touriga Nacional and Touriga Franca) grown in five locations across the Côa Region were considered. Sugar accumulation in grapes, with concentrations between 170 and 230 g l-1, was used from 2014 to 2020 as an indicator of technological maturity conditioned by meteorological factors. The climatic time series were retrieved from the EU Copernicus Service, while sugar data were collected by a non-profit organization, ADVID, and by Sogrape, a leading wine company. The software for calibrating and validating this model framework was the Phenology Modeling Platform (PMP), version 5.5, using Sigmoid and growing degree-day (GDD) models for predictions. The performance was assessed through two metrics: Roots Mean Square Error (RMSE) and efficiency coefficient (EFF), while validation was undertaken using leave-one-out cross-validation. Our findings demonstrate that sugar content is mainly dependent on temperature and air humidity. The models achieved a performance of 0.65

Vineyards and clay minerals: multi-technique analytical approach and correlations with soil properties

Purpose of this research is to quantitatively assess the mineral component of vineyard soils, with particular attention to the mineralogical analysis of clays, which represent an element of high importance in the vineyard culture as well as in general agriculture. An X-ray diffraction (XRD) / thermogravimetric (TG) multi-technique analytical approach was developed, tested on soil samples taken from vineyards around the world. This codified analytical procedure was necessary to obtain precise qualitative and quantitative mineralogical data, globally comparable to distinguish the geopedological identity of the vineyards. Soil samples from vineyards of various locations were analysed, in very different geological conditions. The bulk-rock quantitative phase analysis (QPA) was obtained by the Rietveld method while the detailed composition of the clay-sized fraction was determined by modelling of the oriented X-ray diffraction patterns. The research provided a precise classification of the mineral component of soils, distinguishing the mineral phases of the clays and the so-called mixed-layer clay minerals. We found that the content in mixed layers can be directly correlated with the water retention and the cation exchange capacity ​​of the soil, while the presence of other clayey minerals and phyllosilicates in this research did not affect this CEC parameter, which codes the fertility level of the soils. The study demonstrates that terroir, in particular soils formed in complex or very different geological conditions, can only be effectively interpreted by properly analysing its mineral phases, in particular the mixed-layer clay component. These are characteristic abiotic ecological indicators, which may have specific eco-physiological influences on the plant.

The interplay between grape ripening and weather anomalies – A modeling exercise

Current climate change is increasing inter- and intra-annual variability in atmospheric conditions leading to grapevine phenological shifts as well altered grape ripening and composition at ripeness. This study aims to (i) detect weather anomalies within a long-term time series, (ii) model grape ripening revealing altered traits in time to target specific ripeness thresholds for four Vitis vinifera cultivars, and (iii) establish empirical relationships between ripening and weather anomalies with forecasting purposes. The Day of the Year (DOY) to reach specific grape ripeness targets was determined from time series of sugar concentrations, total acidity and pH collected from a private company in the period 2009-2021 in North-Eastern Italy. Non-linear models for the DOY to reach the specified ripeness thresholds were assessed for model efficiency (EF) and error of prediction (RMSE) in four grapevine cultivars (Merlot, Cabernet Sauvignon, Glera and Garganega). For each vintage and cultivar, advances or delays in DOY to target specified ripeness thresholds were assessed with respect to the average ripening dynamics. Long-term meteorological series monitored at ground weather station by means of hourly air temperature and rainfall data were analyzed. Climate statistics were obtained and for each time period (month, bimester, quarter and year) weather anomalies were identified. A linear regression analysis was performed to assess a possible correlation that may exist between ripening and weather anomalies. For each cultivar, ripeness advances or delays expressed in number of days to target the specific ripening threshold were assessed in relation to registered weather anomalies and the specific reference time period in the vintage. Precipitation of the warmest month and spring quarter are key to understanding the effect of climate change on sugar ripeness. Minimum temperatures of May-June bimester and maximum temperatures of spring quarter best correlate with altered total acidity evolution and pH increment during the ripening process, respectively.

Upscaling the integrated terroir zoning through digital soil mapping: a case study in the Designation of Origin Campo de Borja

homogeneous zones by intersecting several partial zonings of major factors that influence vineyard growth. Each of them follows specific process from their corresponding disciplines. Soil zoning specifically refers to a Soil Resource Inventory map that has traditionally been generated by conventional soil mapping methods. These methods have shortcomings in reaching fine cartographic and categorical details and involve significant expenses, which undermines their applicability. A new framework named Digital Soil Mapping has introduced quantitative models by statistical techniques to establish soil-landscape relationships and is able to provide intensive scale cartography.

In the present study, a microzoning at 1:10.000 scale is generated from an initial zoning, where the conventional soil map with polytaxic map units is replaced by a new one from digital techniques that disaggregates them. The comparison between the zonings considers a quantitative evaluation of capability for each Homogeneous Terroir Unit by means of the Viticultural Quality Index and its categorization based on its distribution by map. The spatial intersection of both maps gives rise to a confusion matrix in which the flows of class variations after the substitution are assessed.

The results show a five-fold increase in the number of Homogeneous Terroir Units identified and a larger differentiation among them, evidenced by a wider range in the capability index distribution. Both elements are accompanied by an increase in the detection of areas of higher potential within previously undervalued uniform zones.These features are a direct effect of the improvements brought by Digital Soil Mapping techniques and would verify the advantages of their implementation in the Integrated Terroir zoning. Eventually, such new highly detailed terroir units would benefit precision viticulture and sustainable management practices.