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…

CONTRIBUTION OF VOLATILE THIOLS TO THE AROMA OF RIESLING WINES FROM THREE REGIONS IN GERMANY AND FRANCE (RHEINGAU, MOSEL, AND ALSACE)

Riesling wines are appreciated for their diverse aromas, ranging from the fruity fresh characters in young vintages to the fragrant empyreumatic notes developed with aging. Wine tasters often refer to Riesling wines as prime examples showcasing terroir, with their typical aroma profiles reflecting the geographical provenance of the wine. However, the molecular basis of the distinctive aromas of these varietal wines from major Riesling producing regions in Europe have not been fully elucidated. In this study, new lights were shed on the chemical characterization and the sensory contribution of volatile thiols to Riesling wines from Rheingau, Mosel, and Alsace. First, Riesling wines (n = 46) from the three regions were collected and assessed for their aroma typicality by an expert panel.

Read More

CHARACTERIZATION OF ENOLOGICAL OAK TANNIN EXTRACTS BY MULTI-ANALYTICAL METHODS APPROACH

Oak tannin extracts are commonly used to improve wine properties. The main polyphenols found in oak wood extracts are ellagitannins¹ that release ellagic acid upon hydrolysis and comprise numerous structures². Moreover, oak tannin extracts contain other compounds giving a complex mixture. Consequently, the official OIV method based on gravimetric analysis of the tannin fraction adsorbed on polyvinylpolypyrrolidone is not sufficient to describe their composition and highlight their chemical diversity.

Read More

INSIGHTS ON THE ROLE OF GENES ON AROMA FORMATION OF WINES

Yeast secondary metabolism is a complex network of biochemical pathways and the genetic profile of the yeast carrying out the alcoholic fermentation is obviously important in the formation of the metabolites conferring specific odors to wine. The aim of the present research was to investigate the relative expression of genes involved in flavor compound production in eight different Saccharomyces cerevisiae strains.
Two commercial yeast strains Sc1 (S.cerevisiae x S.bayanus) and Sc2 (S.cerevisiae) and six indigenous S. cerevisiae strains (Sc3, Sc4, Sc5, Sc6, Sc7, Sc8) isolated during spontaneous fermentations were inoculated in Assyrtiko and Vidiano grape must.

Read More

LARGE-SCALE PHENOTYPIC SCREENING OF THE SPOILAGE YEAST BRETTANOMYCES BRUXELLENSIS: UNTANGLING PATTERNS OF ADAPTATION AND SELECTION, AND CONSEQUENCES FOR INNOVATIVE WINE TREATMENTS

Brettanomyces bruxellensis is considered as the main spoilage yeast in oenology. Its presence in red wine leads to off-flavour due to the production of volatile phenols such as 4-vinylphenol, 4-vinylguaiacol, 4-ethylphenol and 4-ethylguaiacol, whose aromatic notes are unpleasant (e.g. animal, leather, horse or pharmaceutical). Beside wine, B. bruxellensis is commonly isolated from beer, kombucha and bioethanol production, where its role can be described as negative or positive. Recent genomic studies unveiled the existence of various populations.

Read More

MONOSACCHARIDE COMPOSITION AND POLYSACCHARIDE FAMILIES OF LYOPHILISED EXTRACTS OBTAINED FROM POMACES OF DIFFERENT WHITE GRAPE VARIETIES

The recovery of bioactive compounds from grape and wine by-products is currently an important and necessary objective for sustainability. Grape pomace is one of the main by-products and is a rich source of some bioactive compounds such as polyphenols, polysaccharides, fatty acids, minerals and seed oil. Polysaccharides contained in the grape cell wall can be rhamnogalacturonans type II (RG-II), polysaccharides rich in arabinose and galactose (PRAG), mannoproteins (MP), homogalacturonans (HG) and non pectic polysaccharides (NPP).

Read More