GRA Treatment and Rotavirus Infection in Mice

All animal experiments were performed according to the NIH Guidelines for Care and Use of Animals with protocol approval from the Montana State University (Protocol number 2011-44) and Washington State University (Protocol number 00453-004) Institutional Animal Care and Use Committees. Male C57Bl/6, C3H/HeJ, NOD2^−/−, and MyD88^−/− mice were obtained from Jackson Laboratories. Animals were co-housed according to treatment group under SPF conditions, with ad libitum access to food and water. All experiments were performed when animals were eight weeks of age and all groups contained five mice/group. GRA treatments and rotavirus infections were performed as previously described [12] (link). Briefly, mice were administered 50 mg/kg of GRA by oral gavage on day one and then a second time on day three. No adverse effects were observed in GRA treated mice. In experiments where groups included rotavirus infected mice, 10⁵ shedding dose 50 (SD₅₀) of murine rotavirus strain EW was administered by oral gavage on day two. EW does not cause diarrhea or other illness in mice >15 days of age, and infection is measured by antigen shedding in feces. Mice were sacrificed on day eleven by overdose CO₂ inhalation.