Salmonella enterica serovar Typhimurium strain ST14028 was used as the strain for the immune modulator preparation (EBST) preparation. S. Typhimurium strain 14028, aroA- (del-STM0978::KanR; aroA null) was used as the live attenuated bacterial (AroA) vaccine [24] control in the mice studies. The cells were grown overnight at 37°C in Tryptic Soy Broth (TSB), washed twice in PBS, and suspended in PBS at a cell density of approximately 109 CFU/ml. To prepare the heat-killed S. Typhimurium (HKST) preparation, aliquots of the bacterial suspension were heat inactivated at 70°C for 1 h. Six to eight weeks old female C57BL/6J mice (Jacksons Laboratory, Bar Harbor, Maine) used in this study were housed in specific pathogen free conditions and cared for according to Texas A&M University Institutional Animal Care and Use Committee guidelines (Protocol number: 2015–0010). This protocol was approved by the Texas A&M University Institutional Animal Care and Use Committee. For bone marrow harvesting, mice were euthanized by Carbon di oxide asphyxiation causing rapid anesthesia, and death occurring with in 2–3 minutes. For experiments involving organ harvest, cervical dislocation was performed manually that resulted in euthanasia with in approximately 10–15 seconds.

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