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…

OENOLOGICAL POTENTIAL OF AUTOCHTHONOUS SACCHAROMYCES CEREVISIAE STRAINS AND THEIR EFFECT ON THE PRODUCTION OF TYPICAL SAVATIANO WINES

Due to the global demand for terroir wines, the winemaking industry has focused attention on exploiting the local yeast microflora of each wine growing region to express the regional character and enhance the sensory profile of wines such as varietal typicity and aroma complexity. The objective of the present study was to isolate and compare the indigenous strains of Saccharomyces cerevisiae present in different vineyards in the Mesogeia – Attiki wine region (Greece), evaluate their impact on chemical composition and sensory profile of Savatiano wines and select the most suitable ones for winemaking process.

A NEW STRATEGY AND METHODOLOGY FOR THE CHARACTERIZATION OF POLYPHENOLS IN FINING PRECIPITATE

Polyphenols are secondary metabolite widely distributed in plant kingdom such as in fruits, in grapes and in wine. During the winemaking process, polyphenols are extract from the skin and seed of the berries. Fining is an important winemaking step just before bottling which has an impact on wine stabilization and clarification. Most the time, fining agent are animal or vegetal protein while some of them can be synthetic polymer like PVPP or natural origin like bentonite.

METHYL SALICYLATE: A TRENDY COMPOUND MARKER OF ZELEN, A UNIQUE SLOVENIAN VARIETY

The wine market interest for autochthonous varieties, particularly from less known wine regions, has significantly raised in the past few years. In that context, Slovenia, a small country from central Europe with a long winemaking tradition, is getting more and more attention, particularly through its range of unique regional varieties. Among them, Zelen, meaning “green” in Slovene, can only be found in the Vipava valley region, located on the western side of the country, near the border with Italy. When they are young, Zelen wines display very singular aromas reminiscent of rosemary, sage and white fruit. Despite its uniqueness, Zelen wine aromatic typicality is poorly documented in the literature.

WINE FERMENTATION METABOLITES PRODUCED BY TWO TORULASPORA DELBRUECKII STRAINS ISOLATED FROM OKANAGAN VALLEY, BC, CANADA VINEYARDS

Wine aroma is influenced by various factors, from agricultural practices in the vineyard to the enological choices made by winemakers throughout the vinification process. Spontaneous fermentations have a characteristically deeper complexity of aromas when compared to fermentations that have been inoculated with Saccharomyces (S.) cerevisiae because of the diversity of microflora naturally present on grape skins. Non-Saccharomyces yeast are being extensively studied for their ability to positively contribute to wine aroma and flavour. These yeasts are known to liberate more bound volatile compounds present in grape must than S. cerevisiae through the enzymatic action of β-glucosidases and β-lyases1.

MONITOR SOME KEY PARAMETERS THROUGH THE IMPLEMENTATION OFCONTINUOUS CONTROL SYSTEMS OF THE MUST-WINE DURING MACERATION-FERMENTATION IN RED WINEMAKING TO MANAGE OPERATIONS IN “AUTOMATION”

This study is aimed to develop a complete tool for the winemaker with, complete and targeted “winemaking recipes” that can be adapted to criteria set by the winemaker, such as: grape variety, grape health status, degree of ripening, desired wine, redox status throughout the alcoholic fermentation.
To get such aim, specific sets of experiments using red grape juices from different varieties (Nebbiolo, Barbera, Pinot noir, etc.) collected at different technological and phenolic maturity points, will be held with “automatized 4.0 tanks” equipped with sensors for measuring: redox potential, dissolved oxygen, relative density, temperature, and color in order to collect a sufficient amount of data preparatory to the creation of operating models in the most widely winemaking situations in which the automatized 4.0 tanks “will be able to independently respond” with the right corrective actions (opening/closing aeration valve, execution/block pumping overs , etc.) if the key parameters exceed the limits of the recommended ranges set in the selected recipe.