Gaseous CO2 signal initiates growth of butyric-acid-producing Clostridium butyricum in both pure culture and mixed cultures with Lactobacillus brevis

Abstract

Microbial strains produce numerous volatile substances in the anaerobic conditions of the human intestines. The availability of CO2 is known to be a prerequisite for bacterial growth in general. In experiments with anaerobic Lactobacillus brevis and Clostridium butyricum bacteria in the Portable Microbial Enrichment Unit (PMEU) it was shown that these strains interact; this interaction being mediated by CO2 emission. CO2 promoted clostridial growth in pure cultures and mixed cultures with lactobacilli. The growth of C. butyricum in pure cultures was much delayed or did not start at all without CO2 from outside. Conversely, the onset of growth was provoked by a short (15 min) CO2 burst. In mixed cultures the presence of lactobacilli in equal numbers speeded up the onset of clostridial growth by 10 h. If C. butyricum cultures designated as PMEU 1, 2, and 3 in cultivation syringes were chained by connecting the gas flow thereby allowing the volatiles of the preceding syringe culture to bubble to the next one, the growth started in 20, 10, or 6 h, respectively. This effect of gaseous emissions from other cultures speeding up the bacterial growth initiation was abolished if the gas was passed through sodium hydroxide to remove the CO2. The positive contribution of lactobacilli to the growth of butyric-acid-producing clostridia documented in this simulation experiment with PMEU has in vivo implications and indicates molecular communication between the species. CO2 is a necessary signal for the growth of clostridia, and lactobacilli can promote clostridial growth in mixed cultures where both bacteria grow well with mutual benefit.