Fenugreek (Trigonella foenum-graecum) is used in many parts of the world for diabetes, cardiovasculardisease, and depression. While the mechanism(s) of these actions is not known, it is becoming increasinglyclear that gut microbiota are key players in homeostasis and also mediate true first-pass metabolism of dietarycompounds. Furthermore, the high protein and fiber content of fenugreek seeds is particularly suited to modifyintestinal bacteria and offset the dysbiotic effects of high fat/low fiber Western-style diets. Indeed, data from ourlab shows that fenugreek significantly alters intestinal microbial populations, and reverses key Western diet-induced changes to gut microbiota. Based on these observations, we propose the following hypothesis: Fenugreek induces physiologic resiliency via changes to intestinal microbiotaOur hypothesis is based on the ability of fenugreek to offset diet-induced gut dysbiosis, and on data showingthat fenugreek-shaped microbiota can replicate selected effects of fenugreek. While these data support a rolefor gut bacteria in beneficial responses to fenugreek, key data are needed to confirm and identify mechanismsby which fenugreek-microbiota interactions drive physiologic benefits. First, the impact of gut microbiota on thebeneficial profile of fenugreek needs to be established. Further, whether fenugreek-microbiota interactions alterthe gut metabolome directly via unmasking/generation of otherwise-inaccessible botanical phytochemicals;or indirectly via altered metabolism of Western diets should be resolved. Finally, identification of intestinal andblood-based metabolites that mediate fenugreek-based physiologic resiliency is needed to accelerate thetranslation of these findings. To meet these needs, we have devised a unique experimental approachcombining conventional and germ-free mice, an adaptive microbiome transplantation paradigm, and a series ofcutting-edge in silico analyses to identify microbiome-derived, fenugreek-based metabolites that drivephysiologic resiliency. Specific aim 1 conventionally-housed and germ-free mice to determine how intestinalmicrobiota shape the beneficial effects of fenugreek. Aim 2 will use microbiome transplants to determine ifspecific microbiota are both necessary and sufficient for the beneficial effects of fenugreek, and will determineif fenugreek-microbiota interactions alter the gut metabolome directly via unmasking/generation of otherwise-inaccessible botanical phytochemicals; or indirectly via altered metabolism of Western diets. Aim 3 will identifythe exact metabolites in blood and intestinal fractions that predict neurobehavioral and metabolic impairmentusing validation cohorts and state-of-the art bioinformatic tools. Identification of these metabolites, and thecommensal bacteria responsible for their production, could spearhead the formulation of safe and effectivestrategies to preserve health in the today's modern environment. Furthermore, improved understanding howgut microbiota balance the interactions of adverse and beneficial dietary elements could be harnessed topromote physiologic resilience in all patients regardless of diet.