Trypticase soy agar deeps

WT or PLD2^−/− mice were subjected to CLP surgery. 24 h after CLP, peritoneal lavage fluid, peripheral blood, and BALF were collected and cultured overnight on blood–agar base plates (Trypticase Soy Agar Deeps; BD) at 37°C. The lungs, livers, and spleens were also isolated, and 10 mg of each tissue was homogenized in 700 µl of PBS. 50 µl of the tissue homogenate was then cultured overnight on blood–agar plates at 37°C. CFUs were determined as described previously (Kim et al., 2011 (link)).

Male ICR mice at the age of 6 weeks (25–28 g body weight) were used. Procedures for the animal experiments were approved by the Animal Experimentation Committee at Hallym University. Mice were intraperitoneally injected with E. coli (5 × 10⁸). Mice were pretreated subcutaneously with PBS or strychnine (0.4 mg/kg) for 30 min before E. coli injection. At 30 min after E. coli injection, vehicle (PBS) or glycine (30 mg/kg) was subcutaneously injected. Eight hours after E.coli injection, mice were sacrificed, and peritoneal lavage fluid was collected and cultured overnight on blood agar base plates (Trypticase Soy Agar Deeps; BD, Franklin Lakes, NJ, USA) at 37°C and the numbers of CFUs were counted.