Maintenance of S. aureus in Co-culture With P. aeruginosa While Growing as Biofilms.

Bacterial biofilms are found in various environmental niches and are mostly comprised by two or more bacterial species. One such example, are the mixed species bacterial biofilms found in chronic lung infections of cystic fibrosis (CF) patients, which include the Staphylococcus aureus and Pseudomonas aeruginosa bacterial species. S. aureus is one of the CF lung initial colonizers and is assumed to be abrogated when P. aeruginosa becomes established, eliminating its involvement as the infection evolves. Common models used in research do not mimic the actual progression of the mixed species biofilms thus, in this work we developed an in vitro model, where S. aureus biofilms establish prior to the introduction of P. aeruginosa, simulating a state that is phenotypically more similar to the one found in CF lungs. Overall our results demonstrate that S. aureus is not outcompeted, and that timing of inoculation and bacterial concentration affect the final bacterial ratio and quorum sensing related gene expression during the dual species biofilm development.

Interaction of Staphylococcus aureus persister cells with the host when in a persister state and following awakening.

Persister cells, a tolerant cell sub-population, are commonly associated with chronic and recurrent infections. However, little is known about their ability to actually initiate or establish an infection, become virulent and cause pathogenicity within a host. Here we investigated whether Staphylococcus aureus persister cells initiate an infection and are recognized by macrophages, while in a persister cell status, and upon awakening due to exposure to cis-2-decenoic acid (cis-DA). Our results show that S. aureus persister cells are not able to initiate infections in A. thaliana and present significantly reduced virulence towards C. elegans compared to total populations. In contrast, awakened S. aureus persister cells are able to initiate infections in A. thaliana and in C. elegans albeit, with lower mortality than total population. Furthermore, exposure of S. aureus persister cells to cis-DA led to a loss of tolerance to ciprofloxacin, and an increase of the bacterial fluorescence to levels found in total population. In addition, macrophage engulfment of persister cells was significantly lower than engulfment of total population, both before and following awakening. Overall our findings indicate that upon awakening of a persister population the cells regain their ability to infect hosts despite the absence of an increased immune response.