Source: UNIV OF WISCONSIN submitted to NRP
PARTNERSHIP: BIOAVAILABILITY OF PROVITAMIN A CAROTENOIDS AND ZINC FROM BIOFORTIFIED MAIZE AND THE IMPACT ON HUMAN HEALTH
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
ACTIVE
Funding Source
Reporting Frequency
Annual
Accession No.
1029750
Grant No.
2023-67017-39057
Cumulative Award Amt.
$788,000.00
Proposal No.
2022-09450
Multistate No.
(N/A)
Project Start Date
Mar 1, 2023
Project End Date
Feb 28, 2027
Grant Year
2023
Program Code
[A1343]- Food and Human Health
Recipient Organization
UNIV OF WISCONSIN
21 N PARK ST STE 6401
MADISON,WI 53715-1218
Performing Department
(N/A)
Non Technical Summary
Biofortification is the process of increasing the nutritional value of the foods that we eat. Vitamin A and zinc are essential micronutrients that the World Health Organization considers to be short-fall nutrients. This proposal seeks to develop and evaluate the efficacy of conventionally bred biofortified orange corn (maize) with high provitamin A carotenoids and zinc to improve human nutrition. The main objectives of this proposal are to: 1) select and evaluate maize germplasm for increased provitamin A and zinc in Mexico and the US; 2) determine the influence of ß-cryptoxanthin-biofortified maize on zinc bioavailability to hen eggs compared with ß-carotene-biofortified maize and test bioefficacy of the biofortified maize and the nutritionally-enhanced eggs in Mongolian gerbils; and 3) ultimately test the selected high carotenoid/zinc variety in a bioefficacy intervention study in Mexican schoolchildren. We will evaluate the influence of the nutritionally enhanced maize on the microbiome profile in the children consuming the biofortified maize versus a typical maize. The two model animals (hens and gerbils) will demonstrate proof-of-concept for enhanced human nutrition prior to orchestrating the human intervention study in Mexico. This proposal aligns with the improvement of the nutritional profile of maize and hen eggs as more nutritious food for human consumption. This US-Mexico partnership encompasses a soil-to- maize-to-animal-to-human approach to meet the global demand for nutritious foods, which is included as part of the US Government Global Food Security Strategy and the United Nation's Sustainable Development Goals.
Animal Health Component
25%
Research Effort Categories
Basic
50%
Applied
25%
Developmental
25%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
7041510101050%
1021510204025%
2021510108125%
Goals / Objectives
We will select agronomically promising high ß-cryptoxanthin (ßCX) and high zinc (Zn) maize varieties in Mexico and evaluate the elite cultivars in Texas. We will develop and select open-pollinated varieties using elite lines with high concentrations of ßCX and Zn in the grain in Mexico under subtropical conditions. We will further evaluate the selected germplasm lines in the semi-arid to sub-humid climate of Texas in high pH soils that limit bioavailability of Zn to maize. We will conduct field and laboratory screening to select the best varieties with the highest micronutrient content to be used in animal models and a human efficacy study. In the animal studies, we will determine the influence of ßCX-biofortified maize on zinc bioavailability to hen eggs compared with ß-carotene-biofortified maize and test bioefficacy of the biofortified maize and eggs in Mongolian gerbils. Animal models appropriate for carotenoid metabolism are few. This study will determine the difference in Zn bioavailability to eggs from hens fed maize biofortified with ßCX or ß-carotene. We will utilize the egg yolks collected from laying hens to evaluate their vitamin A and Zn nutritional value in gerbils. Provitamin A carotenoids are converted to vitamin A during times of vitamin A depletion in gerbils similarly to humans. Investigating health effects of biofortified maize and eggs are essential for marketing enhanced food products.Finally, we will orchestrate a ßCX- and Zn-biofortified maize intervention study in Mexican schoolchildren to assess vitamin A and Zn status and influence on gut microbiome. We will assess changes in total body vitamin A stores and estimate changes in total liver vitamin A reserves with a randomized intervention study using retinol isotope dilution techniques. We will evaluate the impact of biofortified maize feeding on Zn status using changes in serum concentrations. We will evaluate changes in fecal microbiome in a subset of the children between the biofortified maize and negative control groups.
Project Methods
Methods: We will use traditional plant breeding techniques to optimize zinc in orange maize cultivars. We will test food products maize from this nutirionally enhanced maize in animal models. We will finally assess the improvement in vitamin A and zinc status in children using state of the art techniques.Efforts: The maize germplasm will be grown in Mexico and Texas. We will hold field days during the growing season and harvest to demonstrate the nutritional benefits of maize to graduate and undergraduate students. We will orchestrate the animal studies in Wisconsin to show proof of concept while involving undergraduates in the research. Finally, we will orchestrate a maize feeding intervention trial in Mexican schoolchildren that will involve graduate students from both Mexico and Wisconsin.Evaluation: Our studies will be evaluated on many different levels.Evaluation of the proposed objectives will require many types of data acquisition. This project will generate maize genotype and phenotype data at the International Maize and Wheat Improvement Center (CIMMYT) and Texas A&M. CIMMYT will do preliminary evaluation of the zinc content and carotenoid profiles of the maize using standardized methods. CIMMYT will also do retention studies on the products developed for the human study in Mexico. Soil background properties (e.g., pH, electrical conductivity, overall soil organic carbon, inorganic carbon, and soil nitrogen, soil texture), soil fertility assessments (e.g., Mehlich-3 extraction for P and K analysis and soil nitrate and ammonium), and trace element analysis of soil and plant tissues (Zn, Fe, P, K, Mg, Mn, Ca, etc.,) will be obtained by Texas A&M. Animal data will include growth, organ weights, serum vitamin A, tissue carotenoid, and Zn concentrations. Human studies will have de-identified data on all survey tools. Codes will be stored in a locked cabinet in a locked office at University of Sonora. All training and outreach materials will be stored and disseminated as needed. Isotope enrichment data will only have codes associated with them at University of Wisconsin-Madison (UW). All animal and human data will be evaluated by a biostatistician at UW familiar with such study designs.

Progress 03/01/24 to 02/28/25

Outputs
Target Audience:During this period our target audience has been maize breeders. Dr. Tanumihardjoworked with Torbert Rocheford, Purdue University, and Dr. Howe worked withSeth Murray, Texas A&M to grow maize lines within the United States to be used in the upcoming chicken study. At the same time, the Center for Wheat and Maize Improvement under the guidance of Dr. Palacios-Rojas has worked with local maize breeders in Mexico to provide the remaining maize lines. Changes/Problems:We have not made any major changes to our objectives. We are confident that we will be able to orchestrate the chicken study this year. We did substitute one of the experimental arms with a high xanthophyll variety that is currently in development at Purdue under collaborator Rocheford. This enhances the science and does not deter from our objectives. What opportunities for training and professional development has the project provided?We have identified three graduate students and three undergraduate students to assist with these projects. One graduate student was enrolled in University of Sonora to work on his PhD. One student was assigned to this project at Texas A&M to work in the field last summer. Another student was enrolled at the University of Wisconsin-Madison and is currently working on another human study with hopes to transition to this project. Three undergraduates are currently working in Dr. Tanumihardjo's laboratory and learning analytical methods and human research requirements. How have the results been disseminated to communities of interest?We currently do not have complete results from these studies to disseminate. However, during this period we went through the APHIS application process in order to import the maize. We now have permission to import the maize and we can hand-carry it in if we go through the appropriate procedures that have been outlined by USDA. What do you plan to do during the next reporting period to accomplish the goals?We will be using maize lines that have been grown in a chicken study. We will start working on recipe development for the maize studies in Mexican children.

Impacts
What was accomplished under these goals? We were able to grow maize for the chicken study in Mexico, Texas, and Indiana. All of the maize have enhanced xanthophyll content and the we have genotypes in Mexico with enhanced zinc. Two maize lines that were biofortified with β-crytoxanthin were provided by Co-PI Natalia Palacios at CIMMYT in Mexico to co-PI Howe. These lines were planted in early spring 2024 along with a commercial line commonly produced in Texas. These lines were treated with no supplemental zinc or two different supplementation foliar rates. Corn was hand-harvested in fall of 2024 and is currently being shucked and thrushed. Seed will be analyzed for Zn and remainder will be sent to PI Tanumihardjo for carotenoid analysis and use in subsequent experiments. At theUniversity of Sonora, the graduate student analyzed the maize that we had in storage with enhanced beta-cryptoxanthin. The levels were still good. We intend to start working on recipe development for the studies in children.

Publications


    Progress 03/01/23 to 02/29/24

    Outputs
    Target Audience:During this first year, we have been nurturing the partnership among the co-project directors. This has included extensive contract negotiations among the institutions. Therefore, our target audience has been the administrators and scientists that will be assisting with the project for the next four years. This has included maize breeders and nutritionists working together. Changes/Problems:We have not changed any of the approaches yet in the science. This year was spent negotiating the subcontracts. University of Wisconsin-Madison was finally successful in receiving all of the paperwork needed for the two Mexican subcontracts. The Texas A&M subcontract was set up quickly in comparison. What opportunities for training and professional development has the project provided?We have recruited one graduate student for the project at the University of Wisconsin-Madison and she will start in June 2024. We also have one Mexican graduate student who would like to join the human feeding trial in Mexico when we are ready to orchestrate that trial. He is going to register at a local University for this year. How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals?We plan to continue to support the maize breeding efforts in Mexico. Grain will be grown for the human and animal studies. Seed will be sent to Texas as needed. Crops that are harvested in Texas may be able to be used for animal studies. We have identified two assistant professors at University of Wisconsin-Madison who have expertise in chicken and mouse microbiome. We now plan to add this piece to the chicken and gerbil studies with their input. Their names are Vanessa Leone in Animal and Dairy Science and Joe Pierre in Nutritional Sciences.

    Impacts
    What was accomplished under these goals? We started working with the Center for Wheat and Maize Improvement (CIMMYT) in Mexico. They were the first of the four institutions to begin the research. We are currently breeding the maize to have high beta-cryptoxanthin (BCX) and zinc. With oversight by co-PD, Natalia Palacios Rojas, two synthetic materials, namely HighBCX synthetic (BCXS1801) and High ProVA/Zinc synthetic (SYNPROVAZINC), wereutilized for grain production in the main maize cycle in Mexico. The harvest yielded 22 Kg and 19 Kg, respectively, for BCX and zinc synthetics. Analysis of BCX and zinc content using quick technologies (NIR and XRF) resulted in values lower than anticipated, particularly for zinc (below 15 ppm). To address this, we plan to repeat the analysis using inductively coupled plasma mass spectrometryand reevaluate the growing conditions to gain insights into these results. Once the data was confirmed and funding secured, the harvested grain wasready for immediate shipment. Working with collaborator Seth Murray at Texas A&M, co-PD Julie Howe was able to provide the import permit to CIMMYT. She received the seed and will multiply the seed this year for performing the experiments in Texas in the fields. Dr. Palacios is evalutating what germplasm she will plant this year. We have two meetings for the project. We had one in Mexico. We invited two maize geneticists to help us with "brainstorming". Theseincluded Kevin Pixley, Genetic Resource Manager at CIMMYT, and Torbert Rocheford, Professor at Purdue University. In addition to this face-to-face meeting, Dr. Tanumihardjo continues to meet with the co-project directors individually. We also have a group zoom meeting in early spring 2024 to work out logistics for this year. Our next face-to-face meeting will be in August at CIMMYT. We have been working with Dr. Howe on setting up the zinc analysis for the animal and human samples. We still have to do some trouble shooting on the amount of plasma needed for the zinc analysis in the human samples. We are hoping to have a meeting with Scientist Bryan Gannon and Dr. Howe soon to start evaluating why the human samples did not work. Samples for swine were sent down to Dr. Howe, we will evaluate those soon.

    Publications