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…

IMPACT OF MUST NITROGEN DEFICIENCY ON WHITE WINE COMPOSITION DEPENDING ON GRAPE VARIETY

Nitrogen (N) nutrition of the vineyard strongly influences the must and the wine compositions. Several chemical markers present in wine (i.e., proline, succinic acid, higher alcohols and phenolic compounds) have been proposed for the cultivar Chasselas, as indicators of N deficiency in the grape must at harvest [1]. Grape genetics potentially influences the impact of N deficiency on grape composition, as well as on the concentration of potential indicators in the wine. The goal of this study was to evaluate if the che- mical markers found in Chasselas wine can be extended for other white wines to indicate N deficiency in the grape must.

ENRICHMENT OF THE OENOLOGICAL MALDI-TOF/MS PROTEIN SPECTRA DATABASE FOR RELIABLE OENOLOGICAL YEAST AND BACTERIA IDENTIFICATION

The Matrix Assisted Laser Desorption/Ionization–Time-Of-Flight Mass Spectrometry (MALDI-TOF MS) technology is commonly used in food and medical sector to identify yeast or bacteria species isolated from a nutritive culture media. Since a decade, brewery and oenology industries have been attracted to this method which combines fast analysis times, reliability and low cost of analysis. Briefly, this method is based on the comparison of the MALDI-TOF/MS protein spectra of an isolated colony of yeast or bacteria with those contain in a manufacturer’s reference protein spectra database. Initiated in 2015, the creation of the first oenological mass spectra database has proved to be essential for increase quality of species identification.

CHANGES IN CU FRACTIONS AND RIBOFLAVIN IN WHITE WINES DURING SHORT-TERM LIGHT EXPOSURE: IMPACTS OF OXYGEN AND BOTTLE COLOUR

Copper in white wine can be associated with Cu(II) organic acids (Cu fraction I), Cu(I) thiol species (Cu fraction II), and Cu sulfides (Cu fraction III). The first two fractions are associated with the repression of reductive aromas in white wine, but these fractions gradually decrease in concentration during the normal bottle aging of wine. Although exposure of white wine to fluorescent light is known to induce the accumulation of volatile sulfur compounds, causing light-struck aroma, the influence on the loss of protective Cu fractions is uncertain. Riboflavin is known to be a critical initiator of photochemical reac-tions in wine, but the rate of its decay under short-term light exposure in different coloured bottles and for wine of different oxygen concentrations is not well understood.

WINE SWIRLING: A FIRST STEP TOWARDS THE UNLOCKING OF THE WINE’STASTER GESTURE

Right after the pouring of wine in a glass, a myriad of volatile organic compounds, including ethanol, overwhelm the glass headspace, thus causing the so-called wine’s bouquet [1]. Otherwise, it is worth noting that during wine tasting, most people automatically swirl their glass to enhance the release of aromas in the glass headspace [1]. About a decade ago, Swiss researchers revealed the complex fluid mechanics underlying wine swirling [2]. However, despite mechanically repeated throughout wine tasting, the consequences of glass swirling on the chemical space found in the headspace of wine glasses are still barely known.

S. CEREVISIAE AND O. ŒNI BIOFILMS FOR CONTINUOUS ALCOHOLIC AND MALOLACTIC FERMENTATIONS IN WINEMAKING

Biofilms are sessile microbial communities whose lifestyle confers specific properties. They can be defined as a structured community of bacterial cells enclosed in a self-produced polymeric matrix and adherent to a surface and considered as a method of immobilisation. Immobilised microorganisms offer many advantages for industrial processes in the production of alcoholic beverages and specially increasing cell densities for a better management of fermentation rates.