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…

Influence of a spontaneous cover crop on the vineyard and soil erosion under Mediterranean climate

Sixty five % of the agricultural area of the Basque Country located in the DO Ca Rioja corresponds to vineyards. More than 40% of it has an average slope greater than 10%, which makes it sensitive to erosive processes. Furthermore, it is foreseeable that extreme weather events (storms, hail, extreme heat and cold, etc.) will be favored due to climate change. Cover cropping can mitigate this risk, and therefore the objective of this work is to evaluate the impact that a vegetable cover has on the agronomic behavior of the vineyard, the quality of the grape and soil erosion. For this, a trial has been carried out with a Graciano variety vineyard with a slope between 10% -20% during the years 2020 and 2021. Conventional tillage management in the area has been compared (4-6 passes per year of tillage machinery) versus spontaneous vegetation cover management in the vineyard. This implies not tilling and allowing the grass of the land to colonize the range between the lines of vines, controlling their height through 1-3 mowing passes per year, always trying to affect the surface of the land as little as possible. The vegetative growth, yield and quality of the grape and wine was measured. Furthermore, erosion has been measured using Gerlasch boxes. The yield was lower in the second year of the trial in the cover crop treatment, but erosion was significantly reduced.

Projected changes in vine phenology of two varieties with different thermal requirements cultivated in La Mancha DO (Spain) under climate change scenarios

The aim of this work was to analyze the phenology variability of Tempranillo and Chardonnay cultivars, related to the climatic characteristics in La Mancha Designation of Origin, and their potential changes under climate change scenarios. Phenological dates referred to budbreak, flowering, veraison and harvest were analyzed for the period 2000-2019. The weather conditions at daily time scale, recorded during the same period, were also evaluated. The thermal requirements to reach each of these phenological stages were calculated and expressed as the GDD accumulated from DOY=60. Changes in phenology were projected by 2050 and 2070 taking into account those values and the projected temperatures and precipitation, simulated under two Representative Concentration Pathway (RCP) scenarios –RCP4.5 and RCP8.5– using an ensemble of models. The average phenological dates during the period under study were, April 16th ± 6.6 days and April 5th ± 6.0 days for budbreak, May 31st ± 6.0 days and May 27th ± 5.3 days for flowering, July 26th ± 5.6 days and July 25th ± 5.8 days for veraison, and Ago 23rd ± 10.8 days and Ago 17th ± 9.0 days for harvest, respectively, for Tempranillo and Chardonnay. The projected changes in temperature imply an average change in the maximum growing season (April-August) temperatures of 1.2 and 1.9°C by 2050, and 1.6 and 2.6°C by 2070, under the RCP4.5 and RCP8.5 scenarios, respectively. A reduction in precipitation is predicted, which vary between 15% for 2050 under RCP4.5 scenario and up to 30% by 2070 under RCP8.5. The advance of the phenological dates for 2050, could be of 6, 7, 7, and 8 days for Tempranillo and 4, 6, 6 and 9 days for Chardonnay, respectively for budbreak, flowering, veraison and harvest under the RCP4.5 scenario. Under the RCP8.5 emission scenario, the advance could be up to 30% higher.

Grapevine xylem embolism resistance spectrum reveals which varieties have a lower mortality risk in a future dry climate

Wine growing regions have recently faced intense and frequent droughts that have led to substantial economical losses, and the maintenance of grapevine productivity under warmer and drier climate will rely notably on planting drought-resistant cultivars. Given that plant growth and yield depend on water transport efficiency and maintenance of photosynthesis, thus on the preservation of the vascular system integrity during drought, a better understanding of drought-related hydraulic traits that have a significant impact on physiological processes is urgently needed. We have worked towards this end by assessing vulnerability to xylem embolism in 30 grapevine commercial varieties encompassing red and white Vitis vinifera varieties, hybrid varieties characterized by a polygenic resistance for powdery and downy mildew, and commonly used rootstocks. These analyses further allowed a global assessment of wine regions with respect to their varietal diversity and resulting vulnerability to stem embolism. Hybrid cultivars displayed the highest vulnerability to embolism, while rootstocks showed the greatest resistance. Significant variability also arose among Vitis vinifera varieties, with Ψ12 and Ψ50 values ranging from -0.4 to -2.7 MPa and from -1.8 to -3.4 MPa, respectively. Cabernet franc, Chardonnay and Ugni blanc featured among the most vulnerable varieties while Pinot noir, Merlot and Cabernet Sauvignon ranked among the most resistant. In consequence, wine regions bearing a significant proportion of vulnerable varieties, such as Poitou-Charentes, France and Marlborough, New Zealand, turned out to be at greater risk under drought. These results highlight that grapevine varieties may not respond equally to warmer and drier conditions, outlining the importance to consider hydraulic traits associated with plant drought tolerance into breeding programmes and modeling simulations of grapevine yield maintenance under severe drought. They finally represent a step forward to advise the wine industry about which varieties and regions would have the lowest risk of drought-induced mortality under climate change.

20-Year-Old data set: scion x rootstock x climate, relationships. Effects on phenology and sugar dynamics

Global warming is one of the biggest environmental, social, and economic threats. In the Douro Valley, change to the climate are expected in the coming years, namely an increase in average temperature and a decrease in annual precipitation. Since vine cultivation is extremely vulnerable and influenced by the climate, these changes are likely to have negative effects on the production and quality of wine.
Adaptation is a major challenge facing the viticulture sector where the choice of plant material plays an important role, particularly the rootstock as it is a driver for adaptation with a wide range of effects, the most important being phylloxera, nematode and salt, tolerance to drought and a complex set of interactions in the grafted plant.
In an experimental vineyard, established in the Douro Region in 1997, with four randomized blocs, with five varieties, Touriga Nacional, Tinta Barroca, Touriga Franca and Tinta Roriz, grafted in four rootstocks, Rupestris du Lot, R110, 196-17C, R99 and 1103P, data was collected consecutively over 20 years (2001-2020). Phenological observations were made two to three times a week, following established criteria, to determine the average dates of budbreak, flowering and veraison. During maturation, weekly berry samples were taken to study the dynamics of sugar accumulation, amongst other parameters. Climate data was collected from a weather station located near the vineyard parcel, with data classified through several climatic indices.
The results achieved show a very low coefficient of variations in the average date of the phenophases and an important contribution from the rootstock in the dynamic of the phenology, allowing a delay in the cycle of up to10-12 days for the different combinations. The Principal Component Analysis performed, evaluating trends in the physical-chemical parameters, highlighted the effect of the climate and rootstock on fruit quality by grape varieties.

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.