Fenugreek (Trigonella foenum-graecum) is used in many parts of the world for diabetes, cardiovascular disease, and depression. While the mechanism(s) of these actions is not known, it is becoming increasingly clear that gut microbiota are key players in homeostasis and also mediate true first-pass metabolism of dietary compounds. Furthermore, the high protein and fiber content of fenugreek seeds is particularly suited to modify intestinal bacteria and offset the dysbiotic effects of high fat/low fiber Western-style diets. Indeed, data from our lab 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 microbiota Our hypothesis is based on the ability of fenugreek to offset diet-induced gut dysbiosis, and on data showing that fenugreek-shaped microbiota can replicate selected effects of fenugreek. While these data support a role for gut bacteria in beneficial responses to fenugreek, key data are needed to confirm and identify mechanisms by which fenugreek-microbiota interactions drive physiologic benefits. First, the impact of gut microbiota on the beneficial profile of fenugreek needs to be established. Further, whether fenugreek-microbiota interactions alter the 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 and blood-based metabolites that mediate fenugreek-based physiologic resiliency is needed to accelerate the translation of these findings. To meet these needs, we have devised a unique experimental approach combining conventional and germ-free mice, an adaptive microbiome transplantation paradigm, and a series of cutting-edge in silico analyses to identify microbiome-derived, fenugreek-based metabolites that drive physiologic resiliency. Specific aim 1 conventionally-housed and germ-free mice to determine how intestinal microbiota shape the beneficial effects of fenugreek. Aim 2 will use microbiome transplants to determine if specific microbiota are both necessary and sufficient for the beneficial effects of fenugreek, and will determine if 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 identify the exact metabolites in blood and intestinal fractions that predict neurobehavioral and metabolic impairment using validation cohorts and state-of-the art bioinformatic tools. Identification of these metabolites, and the commensal bacteria responsible for their production, could spearhead the formulation of safe and effective strategies to preserve health in the today's modern environment. Furthermore, impro...