CBBP bacterial strains (Clostridium bolteae, Blautia producta, Bacteroides sartorii, Parabacteroides distasonis) were initially isolated from a mouse colony as described elsewhere (Caballero et al., 2017 (link)). Strains were separately maintained in glycerol stock at −80°C. To generate the inoculum to be administered to mice, bacteria were thawed and 50 μl of stock were spread onto pre-reduced Columbia blood agar plates (BD) at 37°C in anaerobic conditions (Coy Laboratory Products) for 48h. Bacterial lawns were collected by scraping and resuspended in pre-reduced PBS to an OD of ~2, equal amounts of suspensions were mixed for the 4 bacteria, and 200 μl of mix were administered to ampicillin-treated mice by oral gavage.
Mice were administered ampicillin in drinking water (0.5 g/l) starting for 4 days prior to reconstitution with CBBP (with the exception of experiments carried on GF mice, for which amipicillin was not used).
Mice were administered ampicillin in drinking water (0.5 g/l) starting for 4 days prior to reconstitution with CBBP (with the exception of experiments carried on GF mice, for which amipicillin was not used).