For these two strains we re-measured the persister fractions in single antibiotics, as well as in all pairwise combinations of the three antibiotics. We found that the killing dynamics were qualitatively similar to those when using a single antibiotic: all kill curves exhibited biphasic behavior, indicating that at least two subpopulations Birinapant of cells were present (Figure 4). Figure 4 Kill curves in combinations of antibiotics are biphasic and vary between treatments. We used combinations of antibiotics to examine the dynamics of cell killing. These dynamics are
similar to those observed in single antibiotics. A–C: Killing dynamics of all replicate cultures upon treatment of strains SC552 with all pairwise combinations of the three antibiotics. D-F: Killing dynamics of strain SC649. The precise dynamics of this killing in combinations of antibiotics may yield additional insight into how persisters are formed. We briefly outline three general possibilities. BMN 673 order (1) No cells persist when a population is simultaneously treated with antibiotics. This implies that the mechanisms underlying persistence to the two antibiotics are exclusive, and cannot occur within the same cell. (2) The fraction of persistent cells under the combination of antibiotics is approximately multiplicative relative
to the fraction in the two single antibiotics. Although this observation would be consistent with several explanations, the simplest is that the mechanisms of persister formation are independently induced, and occur randomly within the same cell. (3) The fraction of persistent cells under a combination of antibiotics is similar to the fraction observed under treatment with the more lethal antibiotic. 5-Fluoracil mouse Again, although several explanations would be consistent with this, the simplest is that cells that are persistent to the more lethal antibiotic are also persistent to the second. We refer to these
three hypotheses as exclusive, independent, and coincident, respectively. We found that for these two strains, there were no cases in which persister fractions were exclusive. Instead, the persister populations were largely coincident, with the fraction of cells in combinations of antibiotics being similar to the fraction observed in the more lethal antibiotic (Figures 4 and 5). This is consistent with this subset of cells being multidrug tolerant. Thus, although not all persisters are multi-drug tolerant, there appears to be a subset that is. Figure 5 A subset of persister cells is multidrug tolerant. The persister fractions estimated from the killing dynamics are shown for single or combinations of antibiotics. A: strain SC552; B: SC649. For both strains, there is a subset of persisters that appear to be resistant to both antibiotics. Toxin-antitoxin pairs are frequently gained and lost in E.