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…

MAPPING THE CONCENTRATIONS OF GASEOUS ETHANOL IN THE HEADSPACE OF CHAMPAGNE GLASSES THROUGH INFRARED LASER ABSORPTION SPECTROSCOPY

Under standard wine tasting conditions, volatile organic compounds (VOCs) responsible for the wine’s bouquet progressively invade the glass headspace above the wine surface. Most of wines being complex water/ethanol mixtures (with typically 10-15 % ethanol by volume), gaseous ethanol is therefore undoubtedly the most abundant VOC in the glass headspace [1]. Yet, gaseous ethanol is known to have a multimodal influence on wine’s perception [2]. Of particular importance to flavor perception is the effect of ethanol on the release of aroma compounds into the headspace of the beverage [1].

IMPACT OF MANNOPROTEIN N-GLYCOSYL PHOSPHORYLATION AND BRANCHING ON WINE POLYPHENOL INTERACTIONS WITH YEAST CELL WALLS

Yeast cell walls (CWs) may adsorb wine components with a significant impact on wine quality. When dealing with red wines, this adsorption is mainly related to physicochemical interactions between wine polyphenols and cell wall mannoproteins. However, mannoproteins are a heterogeneous family of complex peptidoglycans including long and highly branched N-linked oligosaccharides and short linear O-linked oligosaccharides, resulting in a huge structural diversity.

MODULATION OF YEAST-DERIVED AROMA COMPOUNDS IN CHARDONNAY WINES USING ENCAPSULATED DIAMMONIUM PHOSPHATE TO CONTROL NUTRIENT RELEASE

Yeast-derived aroma compounds are the result of different and complex biochemical pathways that mainly occur during alcoholic fermentation. Many of them are related -but not limited- to the availability of nutrients in the fermentation medium and linked to nitrogen metabolism and biomass produced. Besides, the metabolic phase of yeast also regulates the expression of many enzymes involved in the formation of aroma active compounds. The work investigates the overall effect of continuous supplementation of nutrients during alcoholic fermentation of a grape must on the volatile composition of wines.

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.

ASSESSMENT OF GRAPE QUALITY THROUGH THE MONITORING OFPHENOLIC RIPENESS AND THE APPLICATION OF A NEW RAPID METHOD BASED ON RAMAN SPECTROSCOPY

The chemical composition of grape berries at harvest is one of the key aspects influencing wine quality and depends mainly on the ripeness level of grapes. Climate change affects this trait, unbalancing technological and phenolic ripeness, and this further raises the need for a fast determination of the grape maturity in order to quickly and efficiently determine the optimal time for harvesting. To this end, the characterization of variety-specific ripening curves and the development of new and rapid methods for determining grape ripeness are of key importance.