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…

ANTI-TRANSPIRANT MODULATION OF GRAPE RIPENING: EFFECTS ON MERLOT VINE DEVELOPMENT AND ROSÉ WINE PHENOLIC AND AROMATIC PROFILES

Climate changes are impacting viticultural regions throughout the world with temperature increases being most prevalent.1 These changes will not only impact the regions capable of growing grapes, but also
the grapes that can be grown.2 As temperatures rise the growing degree days increase and with it the sugar accumulation within the berries and subsequent alcohol levels in wine. Consequently, viticultural
practices need to be examined to decrease the levels of sugars.

DETERMINATION OF FREE AMINO ACIDS, AMINO ACID POTENTIAL AND PROTEASE ACTIVITY IN THE LEES AND STILL WINES OF CHAMPAGNE

Prior to winemaking, organic or mineral nitrogen compound concentrations are usually measured in the vineyard and in grape musts. These indicators facilitate vine cultivation decisions, usually through yield or vigor. During vinification, yeast and bacteria metabolize nitrogen compounds in the musts in order to generate biomass. After fermentation, the microorganisms rerelease a part of this nitrogen as soluble compounds into the wines. Another part remains bound in the lees and can be lost during racking. The must’s natural nitrogen quantities, additional supplements during fermentation, and lees contact management enhance the release of nitrogen compounds to the wines. During ageing these nitrogen compounds – primarily the amino acids – are implicated in the generation of odorous compounds such as heterocycles(1).

USE OF 13C CP/MAS NMR AND EPR SPECTROSCOPIC TECHNIQUES TO CHARACTERIZE MACROMOLECULAR CHANGES IN OAK WOOD(QUERCUS PETRAEA) DURING TOASTING

For coopers, toasting process is considered a crucial step in barrel production during which oak wood (Q. petraea) develops several aromatic nuances released to the wine during its maturation. Toasting consists of applying different degrees of heat to a barrel for a specific period. As the temperature increases, thermal degradation of oak wood structure produces a huge range of chemical compounds. Many studies have identified the main key aroma volatile compounds (whisky-lactone, furfural, eugenol, guaiacol, vanillin). However, detailed information on how the chemical structure of oak wood degrades with increasing toasting level is still lacking.

PROGRESS OF STUDIES OF LEES ORIGINATING FROM THE FIRST ALCOHOLIC FERMENTATION OF CHAMPAGNE WINES

Champagne wines are produced via a two-step process: the first is an initial alcoholic fermentation of grape must that produces a still base wine, followed by a second fermentation in bottle – the prise de mousse – that produces the effervescence. This appellation produces non-vintage sparkling wines composed of still base wines assembled from different vintages, varieties, and regions. These base wines, or “reserve wines,” are typically conserved on their fine lies and used to compensate for quality variance between vintages (1). Continuously blending small amounts of these reserve wines into newer ones also facilitates preserving the producer’s “house style.”

EFFECTIVENESS OF APPLIED MATERIALS IN REDUCING THE ABSORPTION OF SMOKE MARKER COMPOUNDS IN A SIMULATED WILDFIRE SCENARIO

Smoke taint (ST) is a grape-wine off-flavour that may occur when grapes absorb volatile phenols (VPs) originating from wildfire smoke (1). ST is associated with the negative sensory attributes such as smoky and ashy notes. VPs are glycosylated in the plant and thus present in both free and bound forms (2; 3). Wildfire smoke has resulted in a decline in grape and wine quality and financial losses which has become a prominent issue for the global wine industry.