Murine Peritonitis/Sepsis Model Evaluation

The in vivo murine peritonitis/sepsis model was set up as previously described [48 (link)]. Briefly, eight NMRI mice per treatment group received an intraperitoneal injection with 5×10⁶ c.f.u. of P. aeruginosa PAO1. One hour after infection the mice were treated with 30 mg kg⁻¹ BW tobramycin, 1 mg kg⁻¹ BW C-30 or a combination of both, and an untreated control group was treated with saline. The clinical condition of the mice was scored with a score from 0 to 6 (0=unaffected, 1=slightly affected, 2=affected, 3=clearly affected, 4=very affected and mouse must be sacrificed, 5=motionless and cold, 6=dead) as previously described [48 (link)]. The number of c.f.u. in the blood and peritoneal fluid 2 and 4 h after treatment was determined via serial dilution and drop plating on modified Conradi-Drigalski agar (10 g l⁻¹ detergent, 1 g l⁻¹ Na₂S₂O₃ H₂O, 0.1 g l⁻¹ bromothymol blue, 9 g l⁻¹ lactose and 0.4 g l⁻¹ glucose, pH8.0; SSI, Denmark).