Quantitative detection of lactic acid bacteria in dried sourdoughs using real-time PCR

A real-time PCR system with 16S rRNA gene-targeted group-specific primers was developed to quantitatively detect lactic acid bacteria (LAB) of the genera Lactobacillus, Leuconostoc, Pediococcus, and Weissella in different types of commercially available dried sourdoughs. Despite a high degree of degradation in the DNA isolated from the doughs, the 341-bp 16S rRNA gene fragment of the sourdough LAB biota could specifically be amplified. For dried sourdoughs, the resulting calculated LAB cell counts were determined to be up to 3.7 × 107 cells/g fresh dough, whereas in non-fermented dough acidifiers, used as a control, the calculated LAB cell counts did not exceed 3.6 × 104 cells/g fresh dough. Moreover, the effect of low pH and/or high lactic acid concentrations prevailing in the doughs on the detectability of LAB cells in spray- and roller-dried sourdoughs by PCR was investigated. Drying of non-acidified sourdoughs still permitted to detect the LAB cells by PCR, whereas drying of acidified doughs reduced the detectable cell counts by approximately 5 (spray dried) and 2 (roller dried) orders of magnitudes. Incubation of acidified doughs for 24 h did not affect the detectability of LAB cells in spray-dried doughs but further reduced the detectable cell counts in roller-dried doughs by additional 2 orders of magnitude.