Abstract
Enteric pathogenic bacteria such as Vibrio cholerae and enteropathogenic Escherichia coli (E. coli) cause life-threatening diarrheal diseases that have afflicted humans for centuries. Understanding the effectors required for intestinal colonization is very important to research on bacteria pathogenesis, and is also important to testing new therapeutics and development of the novel vaccines. Here, we describe the Infant Rabbit Colonization Competition Assay, a variant method of the powerful, nonsurgical animal model reported by Ritchie et al. (2010). In our modified assay, wild type and mutant strains are mixed together and inoculated into 2-day-old New Zealand white rabbits. The competitive index for each mutant measures the colonization capacity of the mutant relative to its wild type parental strain in the gastrointestinal tract. Compared to the traditional Sucking Mice model, the clinical and histologic signs of Vibrio cholerae (V. cholerae)-induced disease of infant rabbits more closely resemble human cholera. The larger input bacteria amount of this model also facilitates high-throughput screens, such as Tn-Seq technology (Fu et al., 2013).
Keywords: Vibrio Cholerae, Enteric Bacteria, Animal Model, Colonization Factor, Competetion Assay
Materials and Reagents
Equipment
Procedure
Representative data
Notes
Recipes
Acknowledgments
This protocol was first described and used in our previous Vibrio cholerae intestinal colonization study (Fu et al., 2013), and was adapted from a precious infant rabbit model, which was described by Ritchie et al. (2010). We thank Dr. Waldor and Dr. Ritchie for helpful suggestions and comments. This work was funded by grants AI-018045 and AI-26289 to J.J.M. from the National Institute of Allergy and Infectious Disease.
References
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