Conventional, Gnotobiotic, and Humanized Microbiota Mouse Models of C. difficile Infection

Area of Science:

• Microbiology
• Immunology
• Gastroenterology

Background:

• Clostridioides difficile infection (CDI) is a major public health concern.
• Murine models are crucial for understanding host-pathogen interactions and microbiota dynamics in CDI.
• Current models often rely on antibiotic-induced gut microbiota disruption for C. difficile colonization.

Purpose of the Study:

• To describe flexible and reproducible mouse models for Clostridioides difficile infection (CDI).
• To provide a protocol adaptable for studying acute disease, long-term colonization, and microbiota-mediated resistance.
• To enable research on defined or humanized microbiota in CDI pathogenesis.

Main Methods:

• Antibiotic pretreatment to disrupt the gut microbiota.
• Controlled challenge with Clostridioides difficile.
• Clinical monitoring and microbial enumeration for disease assessment.
• Microbial reconstitution in germ-free mice for defined microbiota studies.

Main Results:

• Development of adaptable and reproducible mouse models for CDI research.
• Detailed protocol covering antibiotic pretreatment, C. difficile challenge, and monitoring.
• Methods for microbial reconstitution in germ-free mice to study defined microbiota.

Conclusions:

• The described mouse models offer a flexible platform for diverse CDI research.
• The protocol supports studies on acute and chronic CDI, as well as microbiota-host interactions.
• These models enhance reproducibility and minimize environmental contamination in CDI research.