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Applied and Environmental Microbiology, October 2006, p. 6725-6733, Vol. 72, No. 10
0099-2240/06/$08.00+0     doi:10.1128/AEM.01183-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Use of Flow Cytometry To Follow the Physiological States of Microorganisms in Cider Fermentation Processes

Mónica Herrero, Covadonga Quirós, Luis A. García, and Mario Díaz^*

Department of Chemical Engineering and Environmental Technology, University of Oviedo, Oviedo, Spain

Received 22 May 2006/ Accepted 26 July 2006

The flow cytometry (FC) technique used with certain fluorescent dyes (ChemChrome V6 [CV6], DRAQ5, and PI) has proven useful to label and to detect different physiological states of yeast and malolactic bacterium starters conducting cider fermentation over time (by performing sequential inoculation of microorganisms). First, the technique was tested with pure cultures of both types of microorganisms grown in synthetic media under different induced stress conditions. Metabolically active cells detected by FC and by the standard plate-counting method for both types of microorganisms in fresh overnight pure cultures gave good correlations between the two techniques in samples taken at this stage. Otherwise, combining the results obtained by FC and plating during alcoholic and malolactic fermentation over time in the cider-making process, different subpopulations were detected, showing significant differences between the methods. A small number of studies have applied the FC technique to analyze fermentation processes and mixed cultures over time. The results were used to postulate equations explaining the different physiological states in cell populations taken from fresh, pure overnight cultures under nonstress conditions or cells subjected to stress conditions over time, either under a pure-culture fermentation process (in this work, corresponding to alcoholic fermentation) or under mixed-fermentation conditions (for the malolactic-fermentation phase), that could be useful to improve the control of the processes.

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* Corresponding author. Mailing address: Department of Chemical Engineering and Environmental Technology, Faculty of Chemistry, University of Oviedo, C/Julián Clavería s/n, 33071 Oviedo, Spain. Phone: 34 985 103439. Fax: 34 985 103434. E-mail: mariodiaz{at}uniovi.es.

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Applied and Environmental Microbiology, October 2006, p. 6725-6733, Vol. 72, No. 10
0099-2240/06/$08.00+0     doi:10.1128/AEM.01183-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

This article has been cited by other articles:

• Quiros, C., Herrero, M., Garcia, L. A., Diaz, M. (2009). Quantitative Approach to Determining the Contribution of Viable-but-Nonculturable Subpopulations to Malolactic Fermentation Processes. Appl. Environ. Microbiol. 75: 2977-2981 [Abstract] [Full Text]  
• Quiros, C., Herrero, M., Garcia, L. A., Diaz, M. (2007). Application of Flow Cytometry to Segregated Kinetic Modeling Based on the Physiological States of Microorganisms. Appl. Environ. Microbiol. 73: 3993-4000 [Abstract] [Full Text]