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…

OPTIMISATION OF THE AROMATIC PROFILE OF UGNI BLANC WINE DISTILLATE THROUGH THE CONTROL OF ALCOHOLIC FERMENTATION

The online monitoring of fermentative aromas provides a better understanding of the effect of temperature on the synthesis and the loss of these molecules. During fermentation, gas and liquid phase concentrations as well as losses and total productions of volatile compounds can be followed with an unprecedented acquisition frequency of about one measurement per hour. Access to instantaneous production rates and total production balances for the various volatile compounds makes it possible to distinguish the impact of temperature on yeast production (biological effect) from the loss of aromatic molecules due to a physical effect³.

THE ROLE OF CELL WALL POLYSACCHARIDES IN THE EXTRACTION OF ANTHOCYANINS AND TANNINS: RESULTS, PERSPECTIVES OF A MORE POSITIVE CONTRIBUTION

The composition of grape berry cell walls was studied on two grape varieties, two years and two maturation levels at the same time as the extraction of anthocyanins and tannins. The chemical composition of skins, seeds, and pulps, focused on polyphenols and polysaccharides, was compared to the chemical composition in polyphenols after extraction from the skins in model solutions or after wine making of the berries. Polyphenols were mainly characterized by UPLC-MS and HPLC-SEC. Polysaccharides were characterized by analysis of the neutral sugar compositions, and also by the CoMPP (comprehensive micropolymer profiling) analysis, a new method which targets the functional groups of cell wall polysaccharides.

LARGE SURVEY OF THE CHEMICAL COMPOSITION OF WINES RESULTING OF THE PRESSING OF RED WINE MARC. FIRST RESULTS

In the Bordeaux vineyards, press red wine represents about 15% of the volume of wines. Valuing this large volume of press wine is necessary from an economic point of view, of course, but also because of their organoleptic contribution to the blend. Nevertheless, there is a lack of recent knowledge on the composition of press wines. This work aims to establish an initial assessment of their composition (aromatic and polyphenolic) and to set up hypothesis on to the links with their sensorial identity.

CHARACTERIZATION AND IDENTIFICATION OF YEAST BIOACTIVE PEPTIDES RELEASED DURING FERMENTATION AND AUTOLYSIS IN MODEL WINE

Aging wine on lees is a consolidated practice during which some yeast components (e.g., polysaccharides,
proteins, peptides) are released and solubilized in wine thus, affecting its stability and quality.
Apart from the widely studied mannoproteins, the role of other yeast components in modulating wine
characteristics is still scarce. Wine peptides have been studied for their contribution to taste, antioxidant,
and antihypertensive potentials. However, the peptides detected in wine can be influenced by the
interaction between yeasts and grape components.

THE EFFECT OF COPPER ON THE PRODUCTION OF VARIETAL THIOLS DURING THE ALCOHOLIC FERMENTATION OF COLOMBARD AND GROS MANSENG GRAPE JUICES

Nowadays, the rapid growth of vineyards with organic practices and the use of copper as the only fun-gicide against downy mildew raises again the question of the effect of copper on varietal thiols in wine, especially 3-sulfanylhexan-1-ol (3SH) and its acetate (3SHA). A few decades ago, several works indicated that the use of copper in the vineyard had a negative effect on the content of varietal thiols in Sauvignon blanc wines [1, 2]. However, these studies only considered the concentration of the reduced form (RSH) of varietal thiols, without quantifying the oxidised ones. For this purpose, we proposed to monitor both reduced and oxidised forms of varietal thiols in wine under copper stress during alcoholic fermentation to have a more complete picture of the biological and chemical mechanisms.