On days 0 and 1, 4-month-old secondary abiotic IL-10-/- mice (sex-matched littermates; balanced gender ratio) were infected perorally with 109 colony forming units (CFU) of the C. jejuni strain 81–176 grown on Columbia agar (with 5% sheep blood) and selective karmali agar plates (both from Oxoid, Wesel, Germany). Therefore, bacterial colonies were harvested with a sterile swab from respective agar plates after 48-h incubation under microaerophilic conditions and transferred to sterile phosphate-buffered saline (PBS, Thermo Fisher Scientific, Waltham, MA, USA). Immediately thereafter, mice received 0.3 mL of the bacterial suspension by gavage.
As described earlier, the daily pathogen loads were determined in fecal samples after C. jejuni infection and upon necropsy in luminal samples from the gastrointestinal tract (stomach, duodenum, ileum, and colon) by culture [16 (link),38 (link)]. Briefly, serial dilutions of each sample (in sterile PBS, Thermo Fisher Scientific, Waltham, MA, USA) were plated onto karmali agar and incubated under microaerophilic conditions for at least 48 h and at 37 °C. The detection limit of viable pathogens was 100 CFU per g (CFU/g).
As described earlier, the daily pathogen loads were determined in fecal samples after C. jejuni infection and upon necropsy in luminal samples from the gastrointestinal tract (stomach, duodenum, ileum, and colon) by culture [16 (link),38 (link)]. Briefly, serial dilutions of each sample (in sterile PBS, Thermo Fisher Scientific, Waltham, MA, USA) were plated onto karmali agar and incubated under microaerophilic conditions for at least 48 h and at 37 °C. The detection limit of viable pathogens was 100 CFU per g (CFU/g).
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