The effect of subminimal inhibitory concentrations of antimicrobial agents on three bacterial mixtures

A test tube technique was developed to screen bacterial mixtures to detect interbacterial interactions that play a role in determining sensitivity to antimicrobial agents. We found 3 mixtures where these bacterial interactions change the sensitivity to antimicrobials or change the proportions of each bacterial species in the mixture. The mixtures were: Fusobacterium nucleatum 102.3 and Bacteroides endodontalis ATCC 35406; F. nucleatum 102.3 and B. endodontalis BN11 a-f; and Capnocytophaga ochracea 1956c and Eubacterium saburreum 162.4. The antimicrobials used were metronidazole for the first 2 mixtures and tetracycline for the last. F. nucleatum seems to protect B. endodontalis from the action of metronidazole. Conversely, the growth inhibition of C. ochracea by E. saburreum was lifted when tetracycline was present. We also found that the growth of C. ochracea can then permit the subsequent growth of E. saburreum. The test tube method permits the evaluation both of interbacterial interactions and the detection of any protective mechanism against antimicrobial agents in a bacterial mixture. We found that F. nucleatum 102.3 can decrease the metronidazole level in the culture medium, and by the use of 14C-metronidazole we demonstrated that acetamide is produced from metronidazole.