Oral inoculation of TLR2/4 knockout mice

The Institutional Animal Care and Use Committee at the University of Florida approved all mice-related procedures and experiments (IACUC, protocol #201304539). Eight-week old male B6.129-Tlr2^tm1Kir/J and C.C3-Tlr4^Lps-d/J mice were obtained from the Jackson Laboratories (Bar Harbor, ME). Upon arrival, TLR2 and 4 mice were allowed to adapt to the new environment for at least 1 week prior to initiation of any experiment (Rivera et al., 2013 (link)). Mice were housed under specific-pathogen-free facility in University of Florida vivarium. Twenty-four mice were randomly assigned to the infection group and twenty-four to the control group for both TLR2^−/− and TLR4^−/− groups. The infected group orally inoculated by lavage with 10⁹ total cells per mL in RTF-4% CMC as previously described (Rivera et al., 2013 (link), Velsko et al., 2015 (link)), while sham-infected mice were mock-infected with RTF-4% CMC only. At twenty-four weeks of infection, mice were euthanized and sera, jaws and internal organs were collected.