Real-time monitoring method of microbial growth using a simple pressure-based respiration detection system

Abstract

Dry cell weight (DCW) and optical density (OD) measurement methods provide useful data for assessing microbial growth. However, their sampling process is labor-intensive and time-consuming. Therefore, we aimed to evaluate a method for measuring microbial growth through continuous CO₂ measurement under aerobic conditions using a pressure-based respiration detection system, which is traditionally used in anaerobic environments and applies measurement of reduced pressure by capturing CO₂ with KOH. The pressure reduction rate, OD, and DCW values were compared during Ralstonia eutropha H16 culture, which revealed a correlation of R² of 0.99 between the pressure reduction and DCW and a change of DCW (g/L) per pressure (1 mbar) of −0.02 g/L. It showed theoretical limit of detection at 14.67 mbar corresponding to 0.0428 g/L of DCW and theoretical limit of quantification at 48.9 mbar as lower limits. When the pressure-based method was applied to compare carbon source utilization and growth of different strains, such as E. coli sp., Pseudomonas sp., Burkholderia sp., and Bacillus sp., it showed a high correlation with DCW. Overall, these results demonstrate that the pressure-based respiration detection system is a reliable tool for microbial growth monitoring and offers significant advantages by providing real-time data with less labor.