Towards lag phase of microbial populations at growth-limiting conditions: The role of the variability in the growth limits of individual cells

Abstract

The water activity (aw) growth limits of unheated and heat stressed Listeria monocytogenes individual cells were studied. The aw limits varied from 0.940 to 0.997 and 0.951 to 0.997 for unheated and heat stressed cells, respectively. Due to the above variability a decrease in aw results in the presence of a non-growing fraction in the population leading to an additional pseudo-lag in population growth. In this case the total apparent lag of the population is the sum of the physiological lag of the growing cells (time required to adjust to the new environment) and the pseudo-lag. To investigate the effect of aw on the above lag components, the growth kinetics of L. monocytogenes on tryptone soy agar with aw adjusted to values ranging from 0.997 to 0.940 was monitored. The model of B&R was fitted to the data for the estimation of the apparent lag. In order to estimate the physiological lag of the growing fraction of the inoculum, the model was refitted to the growth data using as initial population level the number of cells that were able to grow (estimated from the number of colonies formed on the agar at the end of storage) and excluding the rest data during the lag. The results showed that for the unheated cells the apparent lag was almost identical to the physiological lag for aw values ranging from 0.997 to 0.970, as the majority of the cells in the initial population was able to grow in these conditions. As the aw decreased from 0.970 to 0.940 however, the number of cells in the population which were able to grow, decreased resulting to an increase in the pseudo-lag. The maximum value of pseudo-lag was 13.1h and it was observed at aw=0.940 where 10% of the total inoculated cells were able to grow. For heat stressed populations a pseudo-lag started to increase at higher aw conditions (0.982) compared to unheated cells. In contrast to the apparent lag, a linear relation between physiological lag and aw was observed for both unheated and heat stressed cells.