Murine Models of Klebsiella pneumoniae Gastrointestinal Infection
Juan D Valencia-Bacca1,2, Md Maidul Islam3,2, Karen M Haas1
1Department of Microbiology and Immunology, Wake Forest University School of Medicine, Winston-Salem, North Carolina.
Abstract:
Klebsiella pneumoniae (Kpn) is a Gram-negative opportunistic pathogen and a leading cause of hospital-acquired infections. The gastrointestinal (GI) tract serves as a critical site for Kpn colonization and a reservoir for host-to-host spread that primarily occurs through the fecal-oral route. The emergence of multidrug-resistant strains, coupled with virulence factors such as enhanced capsule production, presents significant challenges for clinical management and public health. Animal models-particularly murine models of GI colonization-are essential for advancing our understanding of Kpn pathogenesis, host-pathogen interactions, and for identifying therapeutic strategies. In this manuscript, we describe standardized protocols for studying Kpn GI colonization and systemic dissemination. These methods are designed to facilitate reproducibility and consistency across laboratories investigating Kpn colonization dynamics, dissemination routes, and immune evasion mechanisms. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. By providing detailed and optimized experimental approaches, this work aims to support the development of uniform methodologies for Kpn GI colonization and host response studies Basic Protocol 1: Murine model of Kpn gastrointestinal colonization via oral feeding Basic Protocol 2: Murine model of Kpn gastrointestinal colonization via gavage Basic Protocol 3: Murine model of Kpn intraperitoneal challenge Basic Protocol 4: Murine model of systemic Kpn infection via intravenous injection Basic Protocol 5: Kpn transmission studies following gastrointestinal colonization with intact microbiota Basic Protocol 6: Transmission studies of Kpn gastrointestinal colonization in antibiotic-pretreated mice Support Protocol 1: Preparation of Kpn inoculum Support Protocol 2: Preparation of mixed Kpn inoculum for competitive infection Support Protocol 3: Assessment of Kpn bacterial burden Support Protocol 4: Preparation of cecal filtrate.
Insights
Standardized protocols for studying Klebsiella pneumoniae (Kpn) gastrointestinal colonization and spread in mice are presented. These methods aim to improve reproducibility in understanding Kpn pathogenesis and developing new treatments.
Area of Science:
- Microbiology
- Infectious Diseases
- Animal Models
Background:
- Klebsiella pneumoniae (Kpn) is a major cause of hospital-acquired infections.
- The gastrointestinal tract is a reservoir for Kpn, facilitating host-to-host spread.
- Multidrug resistance and virulence factors complicate Kpn management.
Purpose of the Study:
- To describe standardized protocols for studying Kpn gastrointestinal colonization and systemic dissemination in murine models.
- To promote reproducibility and consistency in Kpn research.
- To support the development of uniform methodologies for Kpn colonization and host response studies.
Main Methods:
- Detailed protocols for establishing Kpn gastrointestinal colonization in mice via oral feeding or gavage.
- Methods for assessing Kpn systemic dissemination through intraperitoneal challenge or intravenous injection.
- Protocols for Kpn transmission studies, including those with intact or antibiotic-altered microbiota.
Main Results:
- Established standardized experimental approaches for Kpn GI colonization and dissemination.
- Provided detailed methods for inoculum preparation and bacterial burden assessment.
- Included protocols for Kpn transmission studies under various conditions.
Conclusions:
- The described protocols facilitate reproducible research on Kpn pathogenesis and spread.
- These standardized methods are crucial for advancing our understanding of Kpn infections.
- This work supports the development of effective therapeutic strategies against Kpn.
