In vivo commensal control of Clostridioides difficile virulence

Protocol for using negative pressure isolator systems to study BSL-2 organisms in gnotobiotic murine models

“…Original stock concentration (CFU/mL) × stock volume = 5 × 10 3 CFU/mL × total gavage volume required × 1.25 (overage) Note: The working concentration of 5 × 10 3 CFU/mL is based on a 200 μL dose containing 1 × 10 3 CFU used to challenge mice in the reference article for this protocol ( Girinathan et al., 2021 ) Transfer inoculum into a 50 mL conical tube. …”

“…Note: The working concentration of 5 × 10 3 CFU/mL is based on a 200 μL dose containing 1 × 10 3 CFU used to challenge mice in the reference article for this protocol ( Girinathan et al., 2021 )…”

“… CRITICAL: Timing and conditions depend on the characteristics and growth kinetics of the commensal species being studied. We have used 1 × 10 8 CFU per mouse based on the reference article for this protocol ( Girinathan et al., 2021 ). For any new bacterial species, we recommend pilot studies to determine the appropriate concentration to dose GF mice.…”

“…This approach supports experimental systems with defined-association mice and enables high-resolution mechanistic studies of pathogen-commensal interactions and their impacts on host phenotypes. For complete details on the use and execution of this protocol, please refer to Girinathan et al. (2021) .…”

“…For complete details on the use and execution of this protocol, please refer to Girinathan et al. (2021) .…”