Research Project: Evaluation of the Chemical and Physical Properties of Low-Value Agricultural Crops and Products to Enhance Their Use and Value

Location: Functional Foods Research

2020 Annual Report

Objectives
Objective 1: Evaluate and characterize chemicals and nutriceuticals in agricultural crops and products for new or improved food and feed uses. Objective 2: Enhance methodologies to quickly determine and evaluate chemical components in a given agricultural product. Objective 3: Enhance methodologies to rapidly and non-destructively assess the identity and levels of key phytochemicals and nutriceuticals in large sample sets of raw agricultural harvests and products. Objective 4: Evaluate and characterize phytochemical composition and bio-physical properties from mid-west area crops, and under-utilized plants and nuisance plants to develop new materials for use as naturally based bio-pesticides for microbial, insect, or weed pests, or to enhance home garden, organic, agricultural, horticultural plant growth and production. Objective 5: Evaluate and characterize phytochemical composition and biophysical properties to develop new bio-based additives and products for the production of new bio-based ingredients as plastics, fillers, delivery agents, replacement ingredients for the production of new bio-based consumer products.

Approach
The goals of this project are (1) to develop accurate analytical methodology and rapid non-destructive spectrophotometric analytical methods to rapidly assess the levels of specific phytochemicals in seeds, tissues, and processed products, and (2) to develop new uses for low value agricultural waste and co-products, under-utilized plant species. Methodologies will be applied to prepare sufficient quantities of pure phytochemicals for further research, to prepare green-process extracts with defined phytochemical composition, and to characterize the phytochemical composition in products and co-products processed from established crops and new crops, as well as in products from biofuel crops, such as oil seed press cakes, straw, and processing residues. Characterization of key phytochemicals from crude and processed agricultural products and co-products will be used to drive the development of new products from current agricultural crops, as well as developing new and alternative crops. The production of biofuels and agricultural food products generates a variety of co-products (carbon dioxide, sugars, fibers, corn dried distillers grain, glycerol, seed press cakes) and other less valuable residues. Redirecting these wastes to more profitable, higher value uses would benefit both the producers and processors. This project will evaluate and utilize green extraction methods in the preparation of refined phytochemical products, which will be used as ingredients for both functional foods, and new non-food agronomic uses, such as for new uses as functional food ingredients, bio-pesticides and bio-control agents, bio-fillers and additives for alternative bio-fiber and bio-plastic products, and as soil amendments for use in organic farming, lawn care, potting mixes, and home garden products.

Progress Report
This is the final report for this project which terminated in March 2020 and has been replaced by new Project 5010-41000-183-00D, "Development of Enhanced Bio-Based Products from Low Value Agricultural Co-Products and Wastes." The goals of this project are (1) to develop accurate analytical methodology and rapid non-destructive spectrophotometric analytical methods to rapidly assess the levels of specific phytochemicals in seeds, tissues, and processed products, and (2) to develop new uses for low value agricultural waste and co-products, and under-utilized plant species. Characterization of key physical properties and key chemicals from crude and processed agricultural products and co-products will be used to drive the development of new products from current agricultural crops and alternative crops. The use of liquid chromatography-mass spectrometry (LC-MS) as a rapid method for determining phytochemical composition has been developed 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. Major plant natural products have been identified and quantified. 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; flavonoids from broccoli for the bioavailability in rat digestion models as part of work aimed at elucidating synergistic effects of flavonoids with the isothiocyanates found in broccoli; 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. Any chemical analysis is destructive, time consuming, and is not comprehensive. Nondestructive spectral analysis techniques such as near infrared spectrometry (NIRS) are rapid, nondestructive and have been shown to be comprehensive. NIRS has been used for the rapid evaluation of bulk parameters, such as total protein, total fats, total moisture, and total fiber, in a variety of food grains. Working with funding from the United Soybean Board we developed NIRS calibrations for accurate estimates of specific chemical constituents such as the soluble sugar composition and the essential amino acid composition of soybeans. We have found that NIRS does not discriminate the individual soluble sugars or individual amino acids very accurately. However, we have found these methods can be applied to the determination of certain phytochemicals in seeds such as the isoflavones in soybeans. 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, biosolids 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 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. There has been significant interest in using non-petroleum-based adhesives and resins to fabricate wood composites, for both interior and exterior uses. Methodologies were developed using inexpensive by-products from corn, oilseed press cakes, such as milkweed, for production of a more cost-effective bio-based adhesive/resin for this purpose. These products are inexpensive to produce with excellent adhesive and mechanical properties, and can be used in building construction, for furniture components, and in manufacturing industries. As of 2010 the natural fiber composites (NFCs) industry is valued at $2.1 billion and is expected to grow about 10% annually. NFCs are extensively employed in the automotive industry in non-structural applications. Most matrices employed in NFCs are synthetic resins derived from petroleum sources and create environmental problems when disposed. To address this problem bio-based adhesives such as soy and cottonseed meal flours can be employed as adhesives/resins. We have shown that Osage orange seed meal (OOSM) can be used for its adhesive properties as resin matrix in NFCs. It was found that OOSM was as effective as soy flours. Commercial use of OOSM as bio-based adhesives would benefit both NFC manufacturing industries by fabricating a product that is completely bio-degradable and light weight. We have developed collaborations with several researchers and a private company for source materials and to develop new products. Eastern red cedar is an abundant renewable resource and represents a vast potential source of valuable natural products that may serve as natural biocides. Eastern red cedar wood is the source of cedarwood oil, a valuable commodity used for perfumes, insecticides, and repellents. We have designed new efficient methods to extract cedar wood oil from juniper wood for used as a renewable source of natural materials while providing a source of income for farmers and ranchers as well as meet consumer demand for safe products. The enhanced extracts have been tested for a variety of bioactivities. The extracts have been found to impart resistance to wood-decay and termites when they are impregnated into otherwise susceptible wood. The cedarwood oil has also been shown to be repellent to several species of ants, including red imported fire ants and little fire ants. The cedarwood oil is toxic to black-legged ticks, brown dog ticks, lone star ticks, houseflies and several species of mosquitoes. Cedarwood oil is a very safe material towards humans but has great potential for controlling a wide range of economically important pests. We have also investigated the use of a mixture of a derivative of starch and cedarwood oil (CWO) for pressure treating wood to successfully inhibit attack by termites and wood-rot fungi. In addition, the starch/CWO mixture inhibited the absorption of water by the treated wood.

Accomplishments
1. Seed meal adhesives and low value wood fibers for fabricating composite wood panels. Composite wood panels (CWPs) are engineered wood composites including particleboard, medium-density fiberboard, high-density fiberboard, oriented strand board, and plywood employed in building products. ARS scientists in Peoria, Illinois, have shown that effective low-cost adhesives can be obtained from non-commercial woody plants and low value agricultural by-products such as soybean meal, corn distillers grains, and hedgeapple fruit. These adhesives and fibers were employed to fabricate composite wood panels (CWPs) used in engineered wood composites which are important building products. Bio-based CWPs do not shatter in the same way as commercial CWPs and thermal cyclic aging tests showed the long-term durability and performance were comparable to commercially produced CWPs. The bio-based CWPs were found to meet Industrial Standards required for commercial use. Europe, Canada, and USA have strict green building standards so much interest exists in finding inexpensive effective bio-adhesives and materials to satisfy the current and future demands.

2. Rapid nondestructive determination of natural product content of ground soybeans. Rapid analytical methods using near infrared spectrometry (NIRS) are being used to determine general compositional analysis, such as percent protein, oil, fiber, and moisture, in soybeans. Soybeans contain a unique set of bioactive constituents, such as the isoflavones, in small quantities that are implicated in maintaining the health and preventing disease in the animals that consume them. We have developed NIRS methodology to accurately measure soy natural product concentrations from over 2000 samples collected from 6 crop years and from soybeans grown in locations in 14 states. ARS scientists in Peoria, Illinois, developed NIRS analytical methodology to assess natural product concentrations rapidly and accurately on large sets of samples. Accurately measuring the levels of these compounds in ground soybeans will aid in breeding programs and nutritional assessments for beans being brought to the grain elevators and to processing plants.

3. Comparison of the absorption and metabolism of a major flavonoid compound found in broccoli in model animal digestive tracts. Broccoli has a number of interesting constituents that may have an effect on the prevention of the development of chronic diseases in mammals. It is unknown whether flavonoid-sugars are absorbed intact, or whether the sugars are removed in the gut before the flavonoid is absorbed. Using the purified broccoli flavonoid-sugar compound prepared by ARS scientists in Peoria, Illinois, with University of Illinois collaborators in Champaign, Illinois, showed that the flavonoid-sugar is both absorbed intact in a model digestive system, and after the original compound is broken down into the non-sugar form. This research provides a key piece of knowledge to lead to the further assessment of the biological activity of these compounds in mammals.

4. New cat litter formulation developed from low value plant fibers. ARS scientists in Peoria, Illinois, are utilizing low value invasive Eastern red cedar (ERC; Juniperus virginiana L.) wood chips for the development of new absorbents and pest control uses. ERC lumber and sawdust were processed for optimal use as bio-based absorbents which contains the essential oils that have been shown to be repellant and/or toxic to fleas and ticks. An absorbent formulation consisting of 10% ERC biochar, 84% ERC wood fibers, 4% guar gum, and 2% mineral oil had excellent suppression of odors, including the major cat urine odor compound, and had physical and chemical properties equal to or superior to the top three-selling biobased cat litters currently on the market. This formulation had excellent physical and chemical properties for commercial development as a biobased litter and absorbent.

5. New bio-absorbents developed with biochars from low value plant fibers. Biochar produced from Easten red cedar (ERC) wood was found to have very high surface areas compared to most other biochars allowing it to absorb large amounts of water, nutrients, and organic compounds. ARS scientists in Peoria, Illinois, conducted research for its utilization as a slow-release fertilizer for nursery/greenhouse crops when preconditioned with both organic and inorganic fertilizers. When used as a slow-release fertilizer mixed with a peat/vermiculite potting substrate at 10 and 20% (v%/v%), growth of tomato transplants were comparable to a control using commercial slow-release fertilizers. These results indicate this new formulation of biodegradable absorbent will perform as effectively as other commercially available non-biodegradable absorbents in soil supplements.

6. Evaluation of mustard (carinata) seed meals as feed ingredients for hybrid striped bass. Defatted mustard seed meals are a potential source of protein in prepared feeds for farm raised fish, but often contain bitter compounds that can deter feeding. In collaboration with university researchers, ARS scientists in Peoria, Illinois, conducted feeding experiments to determine the maximum level of bitter phytochemicals in cold-pressed carinata seed meals that could be tolerated by hybrid striped bass. Dietary concentrations of carinata meals below 10% did not reduce feed consumption, utilization and growth. This mustard seed meal could be used as an added protein and nutrient source to diets for fish farming operations.

Review Publications
Eller, F.J., Kirker, G.T., Mankowski, M.E., Hay, W.T., Palmquist, D.E. 2020. Effect of burgundy solid extracted from Eastern Red Cedar heartwood on subterranean termites and wood-decay fungi. Industrial Crops and Products. 144:112023. https://doi.org/10.1016/j.indcrop.2019.112023.
Vaughn, S.F., Moser, J.K., Berhow, M.A., Byars, J.A., Liu, S.X., Jackson, M.A., Peterson, S.C., Eller, F.J. 2020. An odor-reducing, low dust-forming, clumping cat litter produced from Eastern red cedar (Juniperus virginiana L.) wood fibers and biochar. Industrial Crops and Products. 147. Article 112224. https://doi.org/10.1016/j.indcrop.2020.112224.
Tisserat, B., Harry-O'kuru, R.E. 2019. Osage orange, honey locust and black locust seed meal adhesives employed to fabricate composite wood panels. Fibers. 7(10):91. http://doi.org/10.3390/fib7100091.
Bantchev, G.B., Vermillion, K.E., Biresaw, G., Berhow, M.A. 2019. Acetylthiostearates – mass spectroscopy and NMR characterization. Journal of Sulfur Chemistry. 41(2):154-169. https://doi.org/10.1080/17415993.2019.1699928.
Kasiga, T., Fey, A.L., Berhow, M.A., Bruce, T.J., Brown, M.L. 2020. Growth, feeding and thyroxine-related responses of hybrid striped (sunshine) bass (Morone chrysops X M. saxatilis) fed de-oiled carinata (Brassica carinata) meal. Aquaculture Nutrition. 26(1):109-122. https://doi.org/10.1111/anu.12971.
Morales-Ramos, J.A., Rojas, M.G., Dossey, A.T., Berhow, M.A. 2020. Self-selection of food ingredients and agricultural by-products by the house cricket, Acheta domesticus (Orthoptera: Gryllidae): A holistic approach to develop optimized diets. PLoS One. 15(1):e0227400. https://doi.org/10.1371/journal.pone.0227400.
Yegin, S., Saha, B.C., Kennedy, G.J., Berhow, M.A., Vermillion, K. 2020. Efficient bioconversion of waste bread into 2-keto-D-gluconic acid by Pseudomonas reptilivora NRRL B-6. Biomass Conversion and Biorefinery. 10(2):545-553. https://doi.org/10.1007/s13399-020-00656-7.
Tisserat, B., Montesdeoca, N., Boddu, V.M. 2020. Accelerated thermal aging of bio-based composite wood panels. Fibers. 8(5):32. https://doi.org/10.3390/fib8050032.
Berhow, M.A., Singh, M., Bowman, M.J., Price, N.P.J., Vaughn, S.F., Liu, S.X. 2020. Quantitative NIR determination of isoflavone and saponin content of ground soybeans. Food Chemistry. 317:126373. https://doi.org/10.1016/j.foodchem.2020.126373.
Hay, W.T., Behle, R.W., Berhow, M.A., Miller, A.C., Selling, G.W. 2020. Biopesticide synergy when combining plant flavonoids and entomopathogenic baculovirus. Nature. 10:6806. https://doi.org/10.1038/s41598-020-63746-6.
Wang, Y., Berhow, M.A., Black, M., Jeffery, E.H. 2019. A comparison of the absorption and metabolism of the major quercetin in brassica, quercetin-3-O-sophoroside, to that of quercetin aglycone, in rats. Food Chemistry. 311:125880. https://doi.org/10.1016/j.foodchem.2019.125880.
Mazewski, C., Luna-Vital, D., Berhow, M., Gonzalez de Mejia, E. 2020. Reduction of colitis-associated colon carcinogenesis by a black lentil water extract through inhibition of inflammatory and immunomodulatory cytokines. Carcinogenesis. 41(6):790-803. https://doi.org/10.1093/carcin/bgaa008.
Kenar, J.A., Felker, F.C., Singh, M., Byars, J.A., Berhow, M.A., Bowman, M.J., Moser, J.K. 2020. Comparison of composition and physical properties of soluble and insoluble navy bean flour components after jet-cooking, soaking, and cooking. LWT - Food Science and Technology. 130. Article 109765. https://doi.org/10.1016/j.lwt.2020.109765.