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Research Project: Plant Antinutrients and the Gut Microbiome: A New Dimension

Location: Children's Nutrition Research Center

2021 Annual Report

Objective 1: Determine if antinutrients in plant foods impact the gut microbiome. Subobjective 1A: Using mice feeding studies, analyze the interaction between antinutrient content in plant-based diets and gut microbiota. Subobjective 1B: Assess the impact of plant-based antinutrient content on animal and human microbiomes using in vitro systems. Objective 2: Utilize a germ-free murine model to determine the inter-relationships between microbial gut ecology, plant anti-nutrients and host mineral bioavailability.

Unique plant diets differing in antinutrient content can be used along with 16S ribosomal RNA sequencing to determine how bacterial populations fluctuate as a function of antinutrients. Calcium (Ca) is often sequestered as an oxalate salt making it an 'antinutrient' and bio-unavailable. Medicago truncatula contains insoluble calcium oxalate crystals making it a poor source of dietary Ca. However, a M. truncatula mutant lacking oxalate crystals in the leaf tissue is an excellent source of Ca and allows the preparation of diets that differ in a single plant mutation to be used to analyze the impact of oxalate on the microbiome. Using primary cell culture models from both humans and mice will provide further insights into the impact of antinutrients on microbial composition. Meanwhile, mice reconstituted with either a microbiome associated with an antinutrient replete or antinutrient deficient diet, but consuming equivalent diets, will be analyzed for differences in calcium, iron, and zinc absorption.

Progress Report
This year we have tried to reestablish plant growth conditions and assays that were paused during the laboratory shutdowns due to COVID-19. We have reached out a company, in Yuma, Arizona, for help growing barley. For Sub-objective 3B, we have successfully established hydroponic growth conditions for the labeling. We have also updated all animal and recombinant DNA protocols associated with this work. We have worked with Collaborators to establish the formulation of the diets. We plan to begin feeding studies in the Fall of 2021.