Determining the Injury-Associated Microbial Nutrients that Facilitate Secondary Bacterial Infection in Acutely Injured Lungs

The goal of this experiment was to determine the changes in the transcriptome of Pseudomonas aeruginosa when cultured in bronchoalveolar lavage fluid (BALF) from mice with oxygen-induced lung injury relative to healthy controls. Overall design: Adult C57BL/6 mice were sacrificed following exposure to hyperoxia (FiO2 95% for 72 hours) or room air (FiO2 21% for 72 hours), BALF was collected and centrifuged to remove host and microbial cells, and stored at -80 C. Upon thawing, BALF was pooled within exposure group, divided into 1 mL aliquots, inoculated with Pseudomonas aeruginosa strain PAO1, and incubated at 37 C shaking to mid-log phase (10 hours). Lennox Broth was used as a positive control for the experiment. Bacterial cell pellets were collected for RNA isolation and bulk RNA-sequencing.