Fluorescence microscopy to monitor wine malolactic fermentation

Malolactic fermentation (MLF) is a natural and biological deacidification of wines and a required step for making premium red wines. MLF is carried out by lactic acid bacteria (LAB) that are present in the fermenting wines. Currently, real-time control of MLF is an issue of great interest as the classical plate count technique for assessing bacterial populations requires long incubation times that are not compatible with a tight control of MLF. The aim of this study was to apply fluorescence microscopy and the bacteria staining kit Live/Dead BacLight (TM) to quantify viable LAB populations in red wines undergoing MLF. This method proved to be a fast and reliable culture-independent method to monitor wine MLF. Moreover, comparison of bacterial population data obtained by fluorescence microscopy and classical plate counts of LAB populations allowed discriminating a population of fully active and culturable cells, from total viable cells that include cells in an intermediate unculturable state.