Establishing C. difficile Infection in HMA Mice

The HMA mice were housed in positive-pressure individually ventilated cages (IVCs) (ISOcage P; Techniplast, West Chester, PA) in order to prevent cross-contamination and maintain their gnotobiotic status ^{49 , 50}. All mice were fed a sterilized rodent breeder diet 5013 (LabDiet, St. Louis, MO). 8- to 16-week-old female and male mice were used in all experiments. All animals were handled in accordance with the protocols approved by the Institutional Animal Care and Use Committee (IACUC) at the University of Michigan. To attain robust C. difficile infection in HC-HMA mice, the animals were treated with a cocktail of antibiotics ofr 5 days. The cocktail contains kanamycin (0.4 mg/ml), gentamicin (0.035 mg/ml), colistin (850 U/ml), metronidazole, (0.215 mg/ml), and vancomycin (0.045 mg/ml). Clindamycin (10 mg/kg) was then injected intra-peritoneally at day 5. One day after clindamycin injection, mice were challenged by C. difficile 24 h post-IP injection ^{17 (link)}.

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Nagao-Kitamoto H., Leslie J.L., Kitamoto S., Jin C., Thomsson K.A., Gillilland MG I.I.I., Kuffa P., Goto Y., Jenq R.R., Ishii C., Hirayama A., Seekatz A.M., Martens E.C., Eaton K.A., Kao J.Y., Fukuda S., Higgins P.D., Karlsson N.G., Young V.B, & Kamada N. (2020). Interleukin-22-mediated host glycosylation prevents Clostridioides difficile infection by modulating the metabolic activity of the gut microbiota. Nature medicine, 26(4), 608-617.

Publication 2020

ISOcage P diet 5013

Animals Antibiotics Clindamycin Colistin Diet Female Gentamicin Infection Institutional animal care and use committee Ip injection Kanamycin Male Metronidazole Mice Pressure Rodent Vancomycin

Corresponding Organization :

Other organizations : University of Michigan–Ann Arbor, University of Gothenburg, Chiba University, The University of Texas MD Anderson Cancer Center, Keio University

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Variable analysis

independent variables

Antibiotic cocktail (kanamycin, gentamicin, colistin, metronidazole, vancomycin)
Clindamycin injection

dependent variables

Establishment of robust C. difficile infection in HC-HMA mice

control variables

HMA mice were housed in positive-pressure individually ventilated cages (IVCs) to prevent cross-contamination and maintain gnotobiotic status
Mice were fed a sterilized rodent breeder diet 5013
8- to 16-week-old female and male mice were used in all experiments
Animals were handled in accordance with the protocols approved by the Institutional Animal Care and Use Committee (IACUC) at the University of Michigan

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