terclim by ICS banner
IVES 9 IVES Conference Series 9 FLOW CYTOMETRY, A POWERFUL AND SUSTAINABLE METHOD WITH MULTIPLE APPLICATIONS IN ENOLOGY

FLOW CYTOMETRY, A POWERFUL AND SUSTAINABLE METHOD WITH MULTIPLE APPLICATIONS IN ENOLOGY

Abstract

Flow cytometry (FCM) is a powerful technique allowing the detection, characterization and quantification of microbial populations in different fields of application (medical environment, food industry, enology, etc.). Depending on the fluorescent markers and specific probes used, FCM provides information on the physiological state of the cell and allows the quantification of a microorganism of interest within a mixed population. For 15 years, the enological sector has shown growing interest in this technique, which is now used to determine the populations present (of interest or spoilage) and the physiological state of microorganisms at the different stages of winemaking.

By studying the other quantification methods now routinely available in enology and comparing them to our FCM method, we show that alternative methods only provide a partial, and sometimes erroneous, view of the microbial populations. Coupled with the use of different markers (vitality markers and probe specific to B. bruxellensis), FCM allows the precise and specific quantification of cells and provides information on their physiological state. In addition, it is the only method that provides a comprehensive view of the present populations, all this in a short time and at a controlled cost.

These advantages make it a method of choice for multiple technical applications in wine microbiology such as the prevention of spoilage during aging, the control of fermentation activity or the quality control of barrel cleaning protocols.

Finally, aside from its technical interest, FCM responds to a major challenge for the wine industry: the commitment to sustainable development. Based on recent work evaluating the environmental impact of analysis techniques, we show through an original comparative study of B. bruxellensis quantification methods that FCM is a more sustainable technique than its alternatives usually used in the sector.

 

1. Longin, C., Julliat, F., Serpaggi, V., Maupeu, J., Bourbon, G., Rousseaux, S., Guilloux-Benatier, M., & Alexandre, H. (2016). Eva-luation of three Brettanomyces; qPCR commercial kits : Results from an interlaboratory study. OENO One, 50(4).
2. Longin C., Laforgue R., Badet-Murat ML., Alexandre H. (2022). Flow cytometry, a sustainable method for the identification and quantification of microorganisms in enology – Part 1/2 Review of the usual methods applied in wine microbiology and the principle of flow cytometry. IVES technical reviews, December 2022.
3. Longin C., Laforgue R., Badet-Murat ML., Alexandre H. (2023). Flow cytometry, a sustainable method for the identification and quantification of microorganisms in enology – Part 2/2 Practical and environmental benefits of flow cytometry applied to wine microbiology. IVES technical reviews, January 2023.
4. Płotka-Wasylka, J. (2018). A new tool for the evaluation of the analytical procedure : Green Analytical Procedure Index. Ta-lanta, 181, 204-209.

DOI:

Publication date: February 9, 2024

Issue: OENO Macrowine 2023

Type: Poster

Authors

Marie-Laure Badet-Murat¹, Cédric Longin¹, Hervé Alexandre²

1. OENOTEAM, 17 Chemin de Verdet, 33500 Libourne/7 Rue de l’Industrie, 33250 Pauillac, France.
2. UMR Procédés Alimentaires et Microbiologiques, Equipe VAlMiS (Vin, Aliment, Microbiologie, Stress), Institut Agro Dijon, Université de Bourgogne/Franche-Comté, IUVV, Rue Claude Ladrey, BP 27877, 21000 Dijon, France.

Contact the author*

Keywords

Brettanomyces bruxellensis, Flow cytometry, Specific quantification, Sustainable analysis

Tags

IVES Conference Series | oeno macrowine 2023 | oeno-macrowine

Citation

Related articles…

PRECISE AND SUSTAINABLE OENOLOGY THROUGH THE OPTIMIZED USE OF AD- JUVANTS: A BENTONITE-APPLIED MODEL OF STUDY TO EXPLOIT

As wine resilience is the result of different variables, including the wine pH and the concentration of wine components, a detailed knowledge of the relationships between the adjuvant to attain stability and the oenological medium is fundamental for process optimization and to increase wine durability till the time of consumption.

EVALUATING WINEMAKING APPLICATIONS OF ULTRAFILTRATION TECHNOLOGY

Ultrafiltration is a process that fractionates mixtures using semipermeable membranes, primarily on the basis of molecular weight. Depending on the nominal molecular weight cut-off (MWCO) specifications of the membrane, smaller molecules pass through the membrane into the ‘permeate’, while larger molecules are retained and concentrated in the ‘retentate’. This study investigated applications of ultrafiltration technology for enhanced wine quality and profitability. The key objective was to establish to what extent ultrafiltration could be used to manage phenolic compounds (associated with astringency or bitterness) and proteins (associated with haze formation) in white wine.

EFFECT OF OXIDATION ON LOW MOLECULAR WEIGHT PHENOLIC FRACTION, SALIVARY PROTEINS PRECIPITATION AND ASTRINGENCY SUBQUALITIES OF RED WINES

Changes in the low molecular weight phenolic fraction, obtained by liquid-liquid microextraction technique, were studied after controlled oxidation of two typologies of Sangiovese wines (Brunello di Montalcino and Chianti Classico) belonging to two vintages (2017 and 2018). The fractions were characterized by LC-MS and quantified by HPLC. The most abundant extracted compounds were the phenolic acids. The effect of oxidation, vintage, and wine typology was stated by a three-ways ANOVA. Gallic and syringic acids significantly increased after oxidation while (–)-epicatechin decreased the most.

IMPACT OF HARVEST DATE ON THE FINE MOLECULAR COMPOSITION OF MUST AND BORDEAUX RED WINE (VAR. MERLOT, CABERNET SAUVIGNON). FOCUS ON ACIDITY AND SENSORY IMPACT AFTER FIVE YEARS OF AGING

Climate change has brought several impacts that are becoming increasingly intense during the last few years and put at risk the quality of the berries or even the plant’s sustainability. Such extreme climatic events impact the composition of the wine while modulating its quality and the consumer preferences (Tempère et al., 2019). The three most important changes that take place in the must are: 1) decrease acidity, 2) increase of the concentration of sugar, hence increase of alcohol in the wine, and 3) modification
of the sensory balance and the development for example of cooked fruit aromas.

EFFECT OF FUMARIC ACID ON SPONTANEOUS FERMENTATION IN GRAPE MUST

Malolactic fermentation (MLF)¹, the decarboxylation of L-malic acid into L-lactic acid, is performed by lactic acid bacteria (LAB). MLF has a deacidifying effect that may compromise freshness or microbiological stability in wines² and can be inhibited by fumaric acid [E297] (FA). In wine, can be added at a maximum allowable dose of 0.6 g/L³. Its inhibition with FA is being studied as an alternative strategy to minimize added doses of SO₂⁴. In addition, wine yeasts are capable of metabolizing and storing small amounts of FA and during alcoholic fermentation (AF).