Rats were placed in an anesthesia chamber with isoflurane and then transferred to an inoculation apparatus, which allowed for continuous general anesthesia during procedure. An otoscope was used as a light source and a special tip was used for visualization of the vocal cords in a method similar to direct laryngoscopy. Once the cords were visualized, 50 μl of 1% bupivicaine was injected for local anesthesia, followed by passage of a catheter through the vocal cords into the trachea and advanced to distal lung tissue. An inoculating syringe was used to deliver 100 μl of the inoculum prepared as described above. Rats were sacrificed at 24 hours and the lung bacterial clearance was quantified by determining the residual for colony-forming units (CFU) of bacteria. Briefly, the entire infected lung was collected and placed in 20 ml of sterile PBS. All inoculated animals only exhibited evidence of gross infection on one side (presumably the site of inoculation), and only that lung that was collected for plating. The unaffected lung did not show bacteria when plated in preliminary experiments and therefore was
