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

Area of Science:

• Microbiology
• Infectious Diseases
• Animal Models

Background:

• Studying host-pathogen interactions requires containment of biological safety level 2 (BSL-2) agents.
• Germ-free and gnotobiotic mouse models offer controlled environments for mechanistic studies.

Purpose of the Study:

• To present a protocol for utilizing negative pressure isolator systems.
• To ensure containment of BSL-2 pathogens in germ-free and gnotobiotic mouse studies.
• To facilitate high-resolution mechanistic studies of pathogen-commensal interactions.

Main Methods:

• Detailed setup and operational procedures for negative pressure isolators.
• Integration of microbiologic procedures within isolator systems.
• Utilizing Clostridioides difficile as a model BSL-2 pathogen.

Main Results:

• Demonstrated a viable protocol for maintaining defined associations in mice.
• Successfully contained BSL-2 pathogens within the isolator systems.
• Enabled mechanistic studies on pathogen-commensal-host interactions.

Conclusions:

• Negative pressure isolators provide a robust system for BSL-2 pathogen research in gnotobiotic mice.
• This approach supports detailed investigation of host phenotypes influenced by microbial communities.
• The protocol facilitates advanced research into infectious disease mechanisms.