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…

Soil, vine, climate change – what is observed – what is expected

To evaluate the current and future impact of climate change on Viticulture requires an integrated view on a complex interacting system within the soil-plant-atmospheric continuum under continuous change. Aside of the globally observed increase in temperature in basically all viticulture regions for at least four decades, we observe several clear trends at the regional level in the ratio of precipitation to potential evapotranspiration. Additionally the recently published 6th assessment report of the IPCC (The physical science basis) shows case-dependent further expected shifts in climate patterns which will have substantial impacts on the way we will conduct viticulture in the decades to come.
Looking beyond climate developments, we observe rising temperatures in the upper soil layers which will have an impact on the distribution of microbial populations, the decay rate of organic matter or the storage capacity for carbon, thus affecting the emission of greenhouse gases (GHGs) and the viscosity of water in the soil-plant pathway, altering the transport of water. If the upper soil layers dry out faster due to less rainfall and/or increased evapotranspiration driven by higher temperatures, the spectral reflection properties of bare soil change and the transport of latent heat into the fruiting zone is increased putting a higher temperature load on the fruit. Interactions between micro-organisms in the rhizosphere and the grapevine root system are poorly understood but respond to environmental factors (such as increased soil temperatures) and the plant material (rootstock for instance), respectively the cultivation system (for example bio-organic versus conventional). This adds to an extremely complex system to manage in terms of increased resilience, adaptation to and even mitigation of climate change. Nevertheless, taken as a whole, effects on the individual expressions of wines with a given origin, seem highly likely to become more apparent.

VINIoT – Precision viticulture service

The project VINIoT pursues the creation of a new technological vineyard monitoring service, which will allow companies in the wine sector in the SUDOE space to monitor plantations in real time and remotely at various levels of precision. The system is based on spectral images and an IoT architecture that allows assessing parameters of interest viticulture and the collection of data at a precise scale (level of grape, plant, plot or vineyard) will be designed. In France, three subjects were specifically developed: evaluation of maturity, of water stress, and detection of flavescence dorée. For the evaluation of maturity, it has been decided first to work at the berry scale in the laboratory, then at the bunch scale and finally in the vineyard. The acquisition of the spectral hyperstal image as well as the reference analyzes to measure the maturity, were carried out in the laboratory after harvesting the berries in a maturity monitoring context. This work focuses on a case study to predict sugar content of three different grape varieties: Syrah, Fer Servadou and Mauzac. A robust method called Roboost-PLSR, developed in the framework of this work (Courand et al., 2022), to improve prediction model performance was applied on spectra after the acquirement of hyperspectral images. Regarding the evaluation of water stress, to work with a significant variability in terms of water status, it has been worked first with potted plants under 2 different water regimes. The facilities have allowed the supervision of irrigation and micro-climatic conditions. The regression models on agronomic variables (stomatal conductance, water potential, …) are studied. To detect flavescence dorée, the experimental plan has consisted of work at leaf scale in the laboratory first, and then in the field. To detect the disease from hyper-spectral imaging, a combination of multivariate curve resolution-alternating least squares (MCR-ALS) and factorial discriminant analysis (FDA) was proposed. This strategy proved the potential towards the discrimination of healthy and infected leaves by flavescence dorée based on the use of hyperspectral images (Mas Garcia et al., 2021).

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.

How does aromatic composition of red wines, resulting from varieties adapted to climate change, modulate fruity aroma?

One of the major issues for the wine sector is the impact of climate change linked to the increasing temperatures which affects physicochemical parameters of the grape varieties planted in Bordeaux vineyard and consequently, the quality of wine. In some varietals, the attenuation of their fresh fruity character is accompanied by the accentuation of dried-fruit notes [1]. As a new adaptive strategy on climate change, some winegrowers have initiated changes in the Bordeaux blend of vine varieties [2]. This study intends to explore the fruitiness in wines produced from grape varieties adapted to the future climate of Bordeaux. 10 commercial single–varietal wines from 2018 vintage made from the main grape varieties in the Bordeaux region (Cabernet franc, Cabernet-Sauvignon and Merlot) as well as from indigenous grape varieties from the Mediterranean basin, such as Cyprus (Yiannoudin), France (Syrah), Greece (Agiorgitiko and Xinomavro), Portugal (Touriga Nacional) and Spain (Garnacha and Tempranillo), were selected among 19 samples using sensory descriptive analyses. Both sensory and instrumental analyses were coupled, to investigate their fruity aroma expression. For sensory analysis, samples were prepared from wine, using a semi preparative HPLC method which preserves wine aroma and isolates fruity characteristics in 25 specific fractions [3,4]. Fractions of interest with intense fruity aromas were sensorially selected for each wine by a trained panel and mixed with ethanol and microfiltered water to obtain fruity aromatic reconstitutions (FAR) [5]. A free sorting task was applied to categorize FAR according to their similarities or dissimilarities, and different clusters were highlighted. Instrumental analysis of the different FAR and wines demonstrated variations in their molecular composition. Results obtained from sensory and gas chromatography analysis enrich the knowledge of the fruity expression of red wines from “new” grape varieties opening up new perspectives in wine technology, including blending, thus providing new tools for producers.

Effects of graft quality on growth and grapevine-water relations

Climate change is challenging viticulture worldwide compromising its sustainability due to warmer temperatures and the increased frequency of extreme events. Grafting Vitis vinifera L.