16Structured abstract word count: (Abstract: 217; Importance: 143)
Abstract 19Pseudomonas aeruginosa and Staphylococcus aureus often cause chronic, recalcitrant infections 20 in large part due to their ability to form biofilms. The biofilm mode of growth enables these 21 organisms to withstand antibacterial insults that would effectively eliminate their planktonic 22 counterparts. We found that P. aeruginosa supernatant increased the sensitivity of S. aureus 23 biofilms to multiple antimicrobial compounds, including fluoroquinolones and membrane-24 targeting antibacterial agents, including the antiseptic chloroxylenol. Treatment of S. aureus with 25 the antiseptic chloroxylenol alone did not decrease biofilm cell viability; however, the 26 combination of chloroxylenol and P. aeruginosa supernatant led to a 4-log reduction in S. aureus 27 biofilm viability compared to exposure to chloroxylenol alone. We found that the P. aeruginosa-28 produced small molecule 2-n-heptyl-4-hydroxyquinoline N-oxide (HQNO) is responsible for the 29 observed heightened sensitivity of S. aureus to chloroxylenol. Similarly, HQNO increased the 30 susceptibility of S. aureus biofilms to other compounds, including both traditional and non-31 traditional antibiotics, which permeabilize bacterial membranes. Genetic and phenotypic studies 32 support a model whereby HQNO causes an increase in S. aureus membrane fluidity, thereby 33 improving the efficacy of membrane-targeting antiseptics and antibiotics. Importantly, our data 34 show that P. aeruginosa exoproducts can enhance the ability of various antimicrobial agents to 35 kill biofilm populations of S. aureus that are typically difficult to eradicate, providing a path for 36 the discovery of new biofilm-targeting antimicrobial strategies. 37 38 Importance 39The thick mucus in the airways of cystic fibrosis (CF) patients predisposes them to frequent, 40 polymicrobial respiratory infections. Pseudomonas aeruginosa and Staphylococcus aureus are 41 frequently co-isolated from the airways of individuals with CF, as well as from diabetic foot 42 ulcers and other wounds. Both organisms form biofilms, which are notoriously difficult to 43 eradicate and promote chronic infection. In this study, we have shown P. aeruginosa secreted 44 factors can increase the efficacy of compounds that alone have little or no bactericidal activity 45 against S. aureus biofilms. In particular, we discovered that P. aeruginosa exoproducts can 46 potentiate the anti-staphylococcal activity of phenol-based antiseptics and other membrane-47 active drugs, including non-traditional antibiotics. Our findings illustrate that polymicrobial 48 interactions can dramatically increase antibacterial efficacy in vitro, and may guide new 49 approaches to target persistent infections, such as those commonly found in respiratory tract 50 infections and in chronic wounds. 51 52
Introduction 53Bacterial biofilms are the underlying cause of many chronic, difficult-to-treat infections. The 54 biofilm lifestyle confers high-level tolerance t...