Research Project: Development of Enhanced Bio-Based Products from Low Value Agricultural Co-Products and Wastes

Location: Functional Foods Research

2021 Annual Report

Objectives
Objective 1: Resolve the unknown biophysical properties of novel bio-based composites and their ingredients to enable commercial fabrication of engineered wood products. Goal 1.1: Identify and develop techniques to convert low value ag-waste (i.e., fermentation residue solids and seed press cakes) and juvenile perennial biomass into marketable commodities. Goal 1.2: Identify and evaluate the factors associated with the response of novel EWP panels to various environmental conditions and methods of their ultimate disposal once their utility function is fulfilled. Objective 2: Convert agricultural wastes and low value byproducts into bio-based pesticides and enhanced soil amendments to increase commercial agricultural and horticultural yields. Goal 2.1: Identify chemical and physical properties of biochars produced from renewable biomass sources and from low-value agricultural co-products and develop these biochars for use as novel, high-value horticultural substrates and for bio based products. Goal 2.2: Evaluate the use of alternative pesticides from a variety of low value plant biomass and from harvesting and processing waste streams. Objective 3: Utilize specific phytochemicals and nutraceuticals from agricultural wastes and low value byproducts to develop new or improve nutritional value in foods and animal feeds. Goal 3.1 Identify key phytochemical components from low value products and wastes to characterize their chemical and biological activities when present alone or in mixtures for determining synergistic properties for new uses as food and feed ingredients. Goal 3.2 Use collaborative studies to determine the role/activities of key phytochemical components for use as bio-pesticides in feeds, feed storage, and plant growth systems. Objective 4: Enhance methodologies to quickly determine and evaluate chemical components and to rapidly and non-destructively assess levels of compositional components in large sample sets of raw agricultural harvests and products. Goal 4.1 Determine if single step accurate mass spectrometric analysis can be used to accurately determine the chemical formulas of phytochemicals present in extracts of seeds, leaves, stems, or bark of several target plant species. Goal 4.2. Determine if accurate NIR calibrations can be obtained for glucosinolate and flavonoid phytochemical components in plant species identified and characterized in the previous research project.

Approach
The overall goal of this project plan is to convert selected low-value agricultural feedstocks into value-added bio-products based upon their physiochemical or chemical properties. The specific bio-products being presented are: (a) engineered wood products (EWP) for indoor uses; (b) biochar as an adaptive for plant growth media; (c) slow-release bio-pesticides; (d) phytochemical (e.g. plant natural essences) based functional food and feed ingredients; and (e) phytochemical based pest control agents. In addition, it is proposed to develop convenient methods for phytochemical discovery and high-throughput methods for measuring amounts of known chemicals present in plant tissues. The feedstocks being investigated include residual pressed oil seed, distillers’ grains with soluble (DDGS) from corn ethanol plants, low-value Midwestern growing trees as well as cedars, and pelletized soybean hulls. Seed cakes will include from soybeans and oil seeds of belonging to the Brassica family that are of emerging interest for industrial applications: Lesquerella, cuphea, and pennycress. One notable aspect of this work is that the combination of feedstock and bioproduct were selected to exploit specific properties of each. The research will also make use of pre-existing expertise in supercritical fluids to develop “green” methods for recovery of bioactive chemicals from plants. Finally, the phytochemical discovery element will be expanded to other crops or plants of emerging interest to further arbitrage newly developed methods.

Progress Report
The overall goal of this project plan is to convert selected low-value agricultural feedstocks into value-added bio-products based upon their physiochemical or chemical properties. The specific bio-products being presented are: (a) engineered manufactured wood for indoor uses; (b) biochar as an adaptive for plant growth media; (c) slow-release bio-pesticides; (d) phytochemical (e.g. plant natural essences) based functional food and feed ingredients; and (e) phytochemical-based pest control agents. Under research for Objective 1: Composite wood panels (CWPs) are employed as building and furniture materials and include plywood, high-density panels, medium-density panels, particle boards. There is much international interest in development of completely biobased CWPs to provide sustainable alternatives to current CWPs which are fabricated with synthetic adhesives. Synthetic adhesives are relatively expensive, non-compostable and are environmentally/chemically hazardous. In contrast, bio-based adhesives are considerably less intrusive to the natural environment and to human health. The majority of biobased adhesives are derived from seed protein flours especially from soybeans. However, these soybean proteins are utilized as a food product which precludes their use as an adhesive because of their cost. In addition, we have continued to study the use of Distiller’s dried grains with solubles (DDGSs) adhesives as substitute to soybean adhesives. DDGSs are considerably less expensive than soybeans and can provide adequate adhesive properties. Under research for Objective 2: Low-value agricultural co-products and bio-solids are being used to make biochar which can be combined with a variety of low-value wastes to create new liquid absorbents and new soil amendments with enhanced plant growth properties. Co-products include harvest residuals and waste products from agricultural processing, such as soybean hulls, bio-solids including municipal sewage solids, and agricultural processing waste streams such as wood sawdust. Biochar is the carbon-rich residual product created under anaerobic conditions by the pyrolysis of plant-based biomass. We determined the addition of processed biosolids and biochars to potting substrates in greenhouse systems and to large-scale turf systems such as golf greens, golf tees, and athletic fields can greatly increase water and nutrient retention, especially in sandy soils. An additional advantage of using biochar instead of other organic amendments is its resistance to microbial decomposition and hence longevity in these applications. We have developed a funded collaboration with the United Soybean Board to examine the use of soy hulls in developing biochar-enhanced absorbents. Eastern red cedar is an abundant renewable resource in the U.S. and represents a vast potential source of valuable natural products that could serve as natural biocides. Cedarwood oil from cedars is toxic to a wide range of economically important arthropods but is very safe for humans. Cedarwood oil has also been demonstrated to impart resistance against both termites and wood-decay fungi in treated wood. Two amylose inclusion complexes were prepared with a primary and a quaternary alkyl amine in combination with burgundy oil from Eastern red cedar which was shown to provide equivalent protection against termites. However, the quaternary alkyl amine was slightly more inhibitory towards wood-decay fungi. Under research for Objectives 3 and 4: The general goal of these objectives is to identify key phytochemical components from low value products and wastes for new uses as food and feed ingredients. We developed the use of liquid chromatography-mass spectrometry (LC-MS) as a rapid method for determining phytochemical composition for a number of plant materials based on collaboration and funding opportunities including: sugar beets, sorghum, soybeans, legume beans and peas, and mustard family plant seeds including broccoli, carinata, and canola. Ongoing work includes the isolation and characterization of saponins from sugar beet processing wastes for use as potential biological control agents of field pests; ferulic acid containing compounds from sorghum stems for elucidating their role in controlling insect pests; anthocyanins from pigmented corn and beans to develop new bioactive food colorants; and volatile isothiocyanates produced from characterized defatted mustard seed meals for use as fungicides. We have also been collaborating with scientists at South Dakota State University to determine if natural antibiotics can be elicited and accumulated in treated soybeans samples for use in animal feeds. Nondestructive spectral analysis techniques such as near infrared spectrometry (NIRS) is rapid and has been shown to be comprehensive. We have shown that NIRS can be applied to the determination of certain phytochemicals in seeds such as the isoflavones in soybeans. We are applying this methodology to other agricultural products. In April of 2020, we were asked by the Office of National Programs to develop an ARS hemp phytochemical analysis protocol. In FY 2020, Congress directed ARS to “conduct regionally driven research, development, and stakeholder engagement to improve agronomic and agro-economic understanding of effectively integrating hemp into existing agricultural cropping, processing, and marketing systems”. This ARS project “Integration of Hemp Production into U.S. Farming Systems” has the objective to provide agronomic information that helps support the incorporation of hemp into farming operations within the U.S. ARS is collaborating with the Global Hemp Innovation Center (Corvallis, Oregon) on ARS research at Peoria, Illinois; Lexington, Kentucky; and Corvallis, Oregon. We began research on postproduction aspects of hemp for use in a variety of new products and applications. Collaboration has been developed to 1) develop consistent and reproducible analytical methods for the analysis of the cannabinoids in hemp samples, and 2) use these methods to measure cannabinoid composition in hemp plant field trials coordinated by the Global Hemp Innovation Center at Oregon State University to develop an understanding of the effect of cultivar and environmental factors on cannabinoid accumulation in hemp. This is going to be integrated with hemp germplasm evaluation work in Oregon, California, and with the hemp breeding program coordinated by the ARS Plant Genetic Resources Lab in Geneva, New York, and the Forage Animal Production Research Unit in Lexington, Kentucky. Work has begun on the evaluation of over 500 samples sent to Peoria from this study. This project received additional funding which was used to hire a post doc and upgrade the mass spectrometry equipment. The team obtained all ARS and U.S. Drug Enforcement Administration permissions and licenses for initiating this research program at Peoria. This new hemp funding supported Peoria ARS researchers to develop and validate a reproducible extraction and measuring procedure to be used worldwide for detecting the total tetrahydrohydrocannabinoid concentrations (total THC: the sum of delta-9 tetrahydrohydrocannabinol (d-9 THC) and tetrahydrohydrocannabinoic acid (THCA) concentrations) in hemp materials, which must be below 0.3% total THC in order to be used as hemp. Meeting this standard is a challenge to U.S. Hemp producers and requires clear and reproducible analytical methodology that does not currently exist. Developing and proving a standard method will assist in the exportation of hemp and hemp products across state and international borders. We now are in the process of running over 500 samples sent from the GHIC utilizing this method. We participated in the National Institute of Standards and Technology cannabinoid evaluation test program with several other labs to examine the reproducibility of analytical methods, the results of which will be published sometime next fiscal year. There is also interest in profiling other plant phytochemicals found in hemp, which includes phenolics, flavonoids, terpenoids, and anthocyanins, and we have initiated that analytical work.

Accomplishments
1. Use of biochar to improve natural potting mix slow-release fertilizer. The majority of components currently used in greenhouse and nursery potting substrates are lightweight materials. However, these materials lack sufficient nutrients for plant growth, so slow-release chemical fertilizers are normally added to supply sufficient nutrients until the plants are placed in gardens. An ARS researcher at Peoria, Illinois, demonstrated that biochar produced from Eastern red cedar wood retained fertilizer if infused with nutrients from synthetic chemical or organic sources, with the potential to release them slowly. Slow-release fertilizers were produced from Eastern red cedar biochar that were infused with either a hydroponic chemical fertilizer or an organic fish-emulsion liquid fertilizer, which are undergoing further evaluation. Eastern red cedar biochar has excellent potential as a slow-release fertilizer for both starter and transplant potting substrates when infused with either inorganic or organic fertilizers and can be used to create new biobased, environmentally-friendly potting mixes with superior performance.

2. Use of dredged waterway sediments for engineered soils for improved bioretention sites along roadways. In order to keep waterways such as rivers and lakes navigable, the United States Army Corps of Engineers performs dredging operations in these bodies of water. However, storage space for the dredged material is limited, so value-added uses for it are needed. A team of ARS researchers at Peoria, Illinois, and collaborators from the Greater Peoria Sanitary District, the U.S. Corps of Engineers, and LHF Compost, Inc. was established in 2018 to study beneficial uses of dredged sediments. Initially, the team has focused on the use of dredged sand from the main channel of the Illinois River to produce engineered soils for use in bioretention sites such as bioswales, rain gardens, and along state highways. We found that grass species used by the Illinois Department of Transportation had excellent growth in engineered soils containing dredged sand and locally produced compost, and good growth if the grasses were grown in engineered soil containing sand, compost, and Greater Peoria Sanitary District biosolids treated to reduce odors, eliminate pathogens and chemical contaminants. The use of dredged material to replace the use of other soil sources for these remediation projects will provide a new use for an underutilized waste material.

3. New biobased litter based on soyhulls. Cats are among the most popular pets in the U.S., and the majority are kept indoors where litter boxes containing some type of absorbent litter material are needed. Multiple companies in the United States are marketing biobased cat litters made of a variety of materials. Biobased litters are attractive because many cat owners are concerned with disposal problems encountered with traditional inorganic clay-based litters. Pet owners fear that cats might harm themselves by ingesting clay litters or inhaling clay dust. ARS researchers at Peoria, Illinois, produced a biobased cat litter based on soybean hulls and soybean hull biochar that had superior odor suppression with much lower dust particle formation compared to three top commercial biobased cat litters, with liquid absorption and clumping ability similar to these commercial litters. A patent application was filed in September 2020 for this formulation to further commercialization of the invention.

Review Publications
Vaughn, S.F., Byars, J.A., Jackson, M.A., Peterson, S.C., Eller, F.J. 2021. Tomato seed germination and transplant growth in a commercial potting substrate amended with nutrient-preconditioned Eastern red cedar (Juniperus virginiana L.) wood biochar. Scientia Horticulturae. 280. Article 109947. https://doi.org/10.1016/j.scienta.2021.109947.
Vaughn, S.F., Theiling, C., Rosenbohm, P., Eller, F.J., Peterson, S.C. 2021. Evaluation of engineered soils for bioretention areas containing dredged Illinois River sand, compost, biosolids and pyrolyzed biosolids. Crop, Forage & Turfgrass Management. 7(1). Article e20096. https://doi.org/10.1002/cft2.20096.
Alhomodi, A.F., Zavadil, A., Berhow, M.A., Gibbons, W.R., Karki, B. 2021. Composition of canola seed sprouts fermented by Aureobasidium pullulans, Neurospora crassa, and Trichoderma reesei under submerged-state fermentation. Food and Bioproducts Processing. 126:256–264. https://doi.org/10.1016/j.fbp.2021.01.008.
Singh, M., Bowman, M.J., Berhow, M.A., Price, N.P., Liu, S.X. 2021. Application of near infrared spectroscopy for determination of relationship between crop year, maturity group, location, and carbohydrate composition in soybeans. Crop Science. 61(4): 2409-2422. https://doi.org/10.1002/csc2.20503.
Alhomodi, A.F., Zavadil, A., Berhow, M.A., Gibbons, W.R., Karki, B. 2021. Application of cocultures of fungal mycelium during solid-state fermentation of canola meal for potential feed application. Journal of the American Oil Chemists' Society. 98(5):509-517. https://doi.org/10.1002/aocs.12479.
Liu, S.X., Chen, D., Plumier, B.M., Berhow, M.A., Xu, J., Byars, J.A. 2020. Impact of particle size fractions on composition, antioxidant activities, and functional properties of soybean hulls. Journal of Food Measurement and Characterization. 15:1547-1562. https://doi.org/10.1007/s11694-020-00746-0.
Cuellar-Núñez, M.L., Loarca-Piña, G., Berhow, M.A., Gonzalez De Mejia, E. 2021. Glucosinolate-rich hydrolyzed extract from Moringa oleifera leaves decreased the production of TNF-alpha and IL-1ß cytokines and induced ROS and apoptosis in human colon cancer cells. Journal of Functional Foods. 75: Article 104270. https://doi.org/10.1016/j.jff.2020.104270.
Hwang, H-S., Winkler-Moser, J.K., Tisserat, B., Harry-O'kuru, R.E., Berhow, M.A., Liu, S.X. 2020. Antioxidant activity of Osage orange extract in soybean oil and fish oil during storage. Journal of the American Oil Chemists' Society. 98(1):73-87. https://doi.org/10.1002/aocs.12458.