Heterogeneity between and within Strains of Lactobacillus brevis Exposed to Beer Compounds

This study attempted to investigate the physiological response of six Lactobacillus brevis strains to hop stress, with and without the addition of Mn²⁺ or ethanol. Based on the use of different fluorescent probes, cell viability and intracellular pH (pHi) were assessed by fluorescence microscopy combined with flow cytometry, at the single cell level. The combined approach was faster than the traditional colony based method, but also provided additional information about population heterogeneity with regard to membrane damage and cell size reduction, when exposed to hop compounds. Different physiological subpopulations were detected under hop stress in both hop tolerant and sensitive strains. A large proportion of cells were killed in all the tested strains, but a small subpopulation from the hop tolerant strains eventually recovered as revealed by pHi measurements. Furthermore, a short term protection against hop compounds was obtained for both hop tolerant and sensitive strains, by addition of high concentration of Mn²⁺. Addition of ethanol in combination with hop compounds caused an additional short term increase in damaged subpopulation, but the subsequent growth suggested that the presence of ethanol provides a slight cross resistance toward hop compounds.