Ampicillin sulbactam

IL-10^−/− mice (in C57BL/6j background, B6) were bred and maintained in the facilities of the “Forschungseinrichtungen für Experimentelle Medizin” (FEM, Charité-Universitätsmedizin, Berlin, Germany) under specific pathogen-free (SPF) conditions.
Gnotobiotic IL-10^−/− mice (with a virtually depleted gastrointestinal microbiota) were generated following broad-spectrum antibiotic treatment as described earlier [31 (link), 36 (link)]. In brief, mice were transferred to sterile cages and treated by adding ampicillin/sulbactam (1 g/L; Pfizer, Berlin, Germany), vancomycin (500 mg/L; Hexal, Holzkirchen, Germany), ciprofloxacin (200 mg/L; Hexal), imipenem (250 mg/L; Fresenius Kabi, Graz, Austria), and metronidazole (1 g/L; Braun, Melsungen, Germany) to the drinking water ad libitum starting at 3 weeks of age immediately after weaning and continued for 3 months before the infection experiment [33 (link)]. Three days prior infection, the antibiotic cocktail was replaced by sterile tap water (ad libitum). Mice were continuously kept in a sterile environment (autoclaved food and tap water) and handled under strict antiseptic conditions.

The spinal contusion was performed using the New York University impactor under isoflurane anesthesia (by a mixture of 4% enflurane and 95% O₂). A laminectomy was performed at the T11 segment of the vertebra and the spinal column was stabilized by clamping the spinous processes of T10 and T12 with the alligator clips. A 10 g weight was dropped onto the spinal cord from a 12.5 mm height (n = 47). After the spinal cord contusion, the musculature was sutured, the skin was autoclipped, and the animals were allowed to recover from the anesthesia. A sham operation (n = 7) was performed using only a laminectomy of the T11 vertebra without a spinal contusion. The bladders were manually expressed twice a day until spontaneous urination returned. In order to prevent a urinary tract infection, Unasyn (ampicillin/sulbactam, 100 mg/kg, Pfizer, Seoul, Korea) was injected intraperitoneally once a day for the first 3 to 5 days. Highly absorbent bedding was used to prevent sores and infections in the paralyzed animals.

IL-10^-/- mice (in C57BL/10 background, B10) were bred and kept in the facilities of the “Forschungseinrichtungen für Experimentelle Medizin” (FEM, Charité - Universitätsmedizin, Berlin, Germany) under specific pathogen-free (SPF) housing conditions. Gnotobiotic IL-10^-/- mice were generated by broad-spectrum antibiotic treatment as described earlier [19 (link)]. In brief, mice were kept in sterile cages and and had ad libitum access to an antibiotic cocktail consisting of ampicillin/sulbactam (1 g/L; Pfizer, Berlin, Germany), vancomycin (500 mg/L; Hexal, Holzkirchen, Germany), ciprofloxacin (200 mg/L; Hexal, Holzkirchen, Germany), imipenem (250 mg/L; Fresenius Kabi, Graz, Austria), and metronidazole (1 g/L; Braun, Melsungen, Germany) in drinking water starting at 3 weeks of age immediately after weaning and continued for 3–4 months before the infection experiment [20 (link)]. Three days prior infection, the antibiotic cocktail was replaced by sterile tap water (ad libitum). These so generated gnotobiotic (i.e. secondary abiotic) mice were continuously kept in a sterile environment (with autoclaved food and drinking water), handeled under strict aseptic conditions and displayed a virtually depleted gastrointestinal microbiota.