Quantitative analysis of Lactobacillus delbrueckii subsp. bulgaricus cell division and death using fluorescent dye tracking

Cell growth of lactic acid bacteria is of vital importance in starter culture manufacture in the dairy industry. However, one of the major stumbling blocks in the understanding of bacterial cell growth relates to challenges in the quantification of the cell division or death. To overcome these shortcomings, the intracellular fluorescent cell tracking assay was implemented to monitor Lactobacillus delbrueckii subsp. bulgaricus sp1.1 cell division and death. Optimization of fluorescent dye concentration suggested it could be applied in the tracking of cell division without cytotoxicity. Technical validation of the fluorescent tracking demonstrated this assay was accurate for quantitatively analyzing cell division. Furthermore, the cell death was quantified using the precursor cohort distribution of the time-series fluorescence data in batch culture. The results indicated a dynamic and unbalanced relationship between bacterial cell division and death after exponential phase in the batch culture. These findings suggested that fluorescent dye tracking is a powerful tool for monitoring L. bulgaricus sp1.1 cell division and death and can provide valuable information for bacterial growth behavior in batch culture.