Changes in antibiotic tolerance are not necessarily predictable a priori. In addition to considering nutrient environment, this data suggests it is critical Pictilisib to know if an antibiotic treatment will be effective over a device’s operational temperature range. 4. AI-2 quorum sensing perturbations Bacteria can communicate with other organisms and can sense properties related to their surroundings using small soluble molecules in a process termed quorum sensing (QS). QS has been associated with the multicellular coordination of many microbial behaviors including pathogenicity and biofilm formation (reviewed in e.g. [21, 22]). Combining QS interference strategies with antibiotic

treatments has been effective against certain microbes under certain conditions and has generated considerable scientific interest (e.g.[23], reviewed in [24]). The efficacy of such combined treatments under perturbed culturing conditions therefore represents a critical assessment of the general applicability of the strategy. A set of E. coli AI-2 QS gene deletion mutants was constructed to act as proxies for QS interference strategies targeting different aspects of AI-2 QS. The Wortmannin solubility dmso strains lacked key enzymes in AI-2 synthesis (ΔluxS), phosphorylation (ΔlsrK), regulation

(ΔlsrR), and degradation LY333531 nmr pathways (ΔlsrF) (reviewed in [25]). The AI-2 system was chosen because of its wide distribution among both Gram negative and positive organisms and because it has been shown to modulate Fossariinae biofilm formation [25]. The E. coli K-12 MG1655 AI-2 QS mutants were constructed using the KEIO gene knock-out library and P1 transduction methods (see materials and methods). The strains were characterized for planktonic and biofilm growth characteristics. Mutant and wild-type planktonic growth rates were nearly identical (Additional file1, Fig. S2). Colony biofilm growth rates and final cell densities also showed no statistical difference (Additional file1, Fig. S3). The AI-2 production profiles for planktonic cultures can be found in Additional

file 1, Fig. S4. The AI-2 profiles were similar to previous reports [26–28]. Perturbation of AI-2 QS did not result in any significant changes in biofilm antibiotic tolerance when cultured at 37°C on LB only medium (Fig. 7a). When the AI-2 QS deletion mutants were perturbed with glucose, non-intuitive changes in antibiotic tolerance were observed. Deleting genes associated with AI-2 synthesis (ΔluxS), regulation (ΔlsrR), or degradation (Δlsrf) increased ampicillin antibiotic tolerance. These cultures had 6 orders of magnitude more cfu’s/biofilm after ampicillin treatment as compared to both wild-type and AI-2 phosphorylation (ΔlsrK) mutants. Additional experimental data regarding the effects of AI-2 gene deletions on antibiotic tolerance can be found in Additional file 1, Figs. S5-S9. Interestingly, the ΔluxS mutant demonstrated an altered glucose catabolite repression response.