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Merck

Citrobacter rodentium mouse model of bacterial infection.

Nature protocols (2016-09-09)
Valerie F Crepin, James W Collins, Maryam Habibzay, Gad Frankel
ABSTRACT

Infection of mice with Citrobacter rodentium is a robust model to study bacterial pathogenesis, mucosal immunology, the health benefits of probiotics and the role of the microbiota during infection. C. rodentium was first isolated by Barthold from an outbreak of mouse diarrhea in Yale University in 1972 and was 'rediscovered' by Falkow and Schauer in 1993. Since then the use of the model has proliferated, and it is now the gold standard for studying virulence of the closely related human pathogens enteropathogenic and enterohemorrhagic Escherichia coli (EPEC and EHEC, respectively). Here we provide a detailed protocol for various applications of the model, including bacterial growth, site-directed mutagenesis, mouse inoculation (from cultured cells and after cohabitation), monitoring of bacterial colonization, tissue extraction and analysis, immune responses, probiotic treatment and microbiota analysis. The main protocol, from mouse infection to clearance and analysis of tissues and host responses, takes ∼5 weeks to complete.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
L-Cysteine, 97%
Sigma-Aldrich
HEPES solution, 1 M, pH 7.0-7.6, sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
Donkey serum
Sigma-Aldrich
Chloramphenicol-Water Soluble, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Brefeldin A, from Penicillium brefeldianum, ≥99% (HPLC and TLC)
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, pH 7, ≥98%
Sigma-Aldrich
Phorbol-12-myristate-13-acetate, Phorbol-12-myristate-13-acetate, CAS 16561-29-8, is the most common phorbol ester. Activates PKC at nanomolar concentrations.