Soybeans have long been used as an important component of production animal diets, particularly in the form of soybean meal (SBM), and serve as a high-quality source of protein with a nearly complete amino acid profile. Outside of its primary nutritive fractions, SBM contains a multitude of minor bioactive components that have variable effects on animal health. Soy saponins are one such component that were previously lumped into the category of anti-nutritional factors found in SBM but have more recently been investigated for their potential anti-inflammatory and anti-oxidative properties. Challenges with their isolation have limited research to in vitro or rodent models, which only require small quantities of purified saponins. However, these studies are less applicable to understanding how saponins are affecting larger production animals that are the primary consumers of SBM. In this research, a process for creating greater quantities of saponins was evaluated for its practicality and effectiveness. A total of 10 kg SBM was used as the starting material for the extraction protocol, which initially contained 5.30 mg total saponins/g SBM. The saponin profile of the meal was predominantly made up of group B saponins, at 57.6% of total saponins, with limited quantities of group A saponins at only 7.17% of the total. The SBM was subjected to two consecutive rounds of mixing with 75% undenatured ethanol at 40℃ for 24 hr. After these mixes, remaining SBM solids were removed and ethanol was evaporated to obtain 12.4 kg of a soy molasses product consisting of the carbohydrate, isoflavone, and saponin fractions of the original SBM. This molasses product was then acidified under elevated temperatures (40℃) using concentrated hydrochloric acid to a pH of 3.05. Under these conditions, a precipitate formed and was recovered via centrifugation to obtain 494 g of precipitated solids likely consisting of isoflavones and saponins. This precipitate was combined with 80% American Chemical Society grade acetone at a 10:1 ratio. The pH of the solution was elevated to 6.00 using sodium hydroxide and the solution was heated to reflux at 56℃ for 90 min. The solution was chilled to 4℃ at which time a precipitate formed and was recovered using vacuum filtration. Analyses are currently being performed to determine the total saponin and isoflavone content as well as the saponin profile of this final product to ensure that saponins were adequately extracted from the initial SBM and isolated from other bioactive components like isoflavones. Pending these results, this large-scale saponin extraction protocol may be utilized to create a saponin-enriched product that can be used in future live animal trials using larger production animals.

Mycotoxin accumulation during the malting of wheat and barley grain causes millions of dollars in annual losses for the U.S. malting and brewing industry. This research investigates the use of biofumigant treatments from plant derived metabolites to reduce grain Fusarium contamination and mycotoxin production during malting. Defatted seed meals from five glucosinolate-containing members of the Brassicaceae Family, Brassica juncea, Brassica carinata, Lepidium sativum, Sinapis alba, and Thlaspi arvense, were used to fumigate inoculated wheat and barley. We found that naturally produced gaseous allyl isothiocyanate was able to control Fusarium graminearum growth and mycotoxin contamination without harming wheat or barley germination. Treatment of Fusarium contaminated barley with volatiles from defatted Brassica juncea, in simulated malting conditions, completely prevented mycotoxin accumulation. No residual isothiocyanates were detected on the malted barley post kilning. This research demonstrates an alternative valorization of Brassicaceae seed meal byproducts as potent inhibitors of fungal contamination during malting.

Fusobacterium necrophorum commonly causes liver abscesses in grain-finished beef, which is mitigated using antibiotics in feed. However, microbial resistance development necessitates exploration of antibiotic alternatives. Here we report the antimicrobial efficacy of Cannabis sativa L. (hemp) crude oil extract and its constituents on Fusobacterium species.

Field pennycress, Thlaspi arvense L., is a new oil seed cover crop being commercialized as a source of biofuel in the United States. Along the path to commercialization, pennycress seed and meal will be marketed as animal feed. As a member of the Brassicacaeae family, pennycress seed is abundant in glucosinolates which are antinutritional factors that make the seed and meal less palatable to animals. More specifically, sinigrin has been identified as the only glucosinolate present in pennycress seed. As work continues to develop varieties low in glucosinolates, a rapid assay is needed to determine the sinigrin level in pennycress seed. The objective of this study was to develop and utilize a high throughput, rapid assay to screen pennycress seeds from wild populations for total sinigrin content. Pennycress seeds were ground in potassium phosphate buffer with endogenous myrosinase to catalyze the enzymatic hydrolysis of sinigrin to equimolar amounts of glucose. Glucose content was determined using a blood glucose meter and test strips. From the molar concentration of glucose, the molar concentration of sinigrin was known and the sinigrin level in seeds was calculated in mu mol/g. The results were confirmed by HPLC/MS analysis. A rapid and inexpensive method was developed and used to screen seeds of 429 wild pennycress lines for sinigrin. Sinigrin levels in seed ranged from 25 to 85 mu mol/g in the pennycress lines screened. Environmental factors also seemed to contribute to the levels of sinigrin in seeds found within the line 'tt8-t/ARV1 ' grown at ten different locations in the Midwest, USA. This rapid and inexpensive assay will allow breeders to efficiently screen lines and select for varieties that produce seeds with low sinigrin levels for improved feed value.

Dehulled peas (DHP) are increasingly popular in food applications, but their integration in food products is limited due to high starch and fiber causing gelling and aggregation during cooking. Additionally, DHP contains saponins (secondary metabolite), contributing to bitter flavors, yet they also hold health benefits. We hypothesize that fungal fermentation could enhance DHP nutritional profile and integration into food products. This study evaluated the effects of six fungal organisms (Aspergillus niger [An], Aspergillus oryzae [Ao], Aureobasidium pullulans [Ap], Neurospora crassa [Nc], Rhizopus microspores var. oligosporus [Ro], Trichoderma reesei [Tr]) on DHP over 120 h of submerged fermentation, evaluating total phenolics, starch, saponins, crude proteins, and overall mass balance. Results from the study demonstrated notable changes post-fermentation, including increased overall protein content and solubility, decreased starch content, reduced overall mass recovery, and elevated levels of total phenolics and saponins. Filamentous fungi exhibited a significant reduction in starch content, contributing to a substantial reduction in mass recovery (31%-60%) compared to the control. Unexpectedly, saponin concentrations increased (1.5 to 3 folds) during fermentation, possibly attributed to the breakdown of the substrate matrix and release of bound saponins. Total phenolic levels varied among microorganisms, with An and Nc demonstrating the highest increases (6 to 10 folds) as compared to the control. Overall, these findings point to fungal fermentation as a tool for adding value to yellow peas and other crops facing similar processing challenges. Further research is warranted to understand the health impacts and value of these enhancements.

Amino acid-fermenting Clostridia have undesirable effects in agricultural systems, which can be mitigated by antibiotics, but resistance necessitates alternatives. Here, we demonstrate the efficacy of cannabidiol on growth and ammonia inhibition of five agriculturally relevant Clostridia: Clostridium sporogenes, Peptostreptococcus spp., Clostridioides difficile, Acetoanaerobium sticklandii, and Clostridium aminophilum.

Strategic, sustainable, and ecofriendly alternatives to chemical pesticides are needed to effectively control mosquitoes and reduce the incidence of their vectored diseases. We evaluated several Brassicaceae (mustard family) seed meals as sources of plant derived isothiocyanates produced from the enzymatic hydrolysis of biologically inactive glucosinolates for the control of Aedes aegypti (L., 1762). Five defatted seed meals (Brassica juncea (L) Czern., 1859, Lepidium sativum L., 1753, Sinapis alba L., 1753, Thlaspi arvense L., 1753, and Thlaspi arvense-heat inactivated and three major chemical products of enzymatic degradation (allyl isothiocyanate, benzyl isothiocyanate and 4-hydroxybenzyl isothiocyanate) were assayed to determine toxicity (LC50) to Ae. aegypti larvae. All seed meals except the heat inactivated T. arvense were toxic to mosquito larvae. L. sativum seed meal was the most toxic treatment to larvae (LC50 = 0.04 g/120 mL dH2O) at the 24-h exposure. At the 72-h evaluation, the LC50 values for B. juncea, S. alba and T. arvense seed meals were 0.05, 0.08 and 0.1 g/120 mL dH2O, respectively. Synthetic benzyl isothiocyanate was more toxic to larvae 24-h post treatment (LC50 = 5.29 ppm) compared with allyl isothiocyanate (LC50 = 19.35 ppm) and 4-hydroxybenzyl isothiocyanate (LC50 = 55.41 ppm). These results were consistent with the higher performance of the benzyl isothiocyanate producing L. sativum seed meal. Isothiocyanates produced from seed meals were more effective than the pure chemical compounds, based on calculated LC50 rates. Using seed meal may provide an effective method of delivery for mosquito control. This is the first report evaluating the efficacy of five Brassicaceae seed meals and their major chemical constituent against mosquito larvae and demonstrates how natural compounds from Brassicaceae seed meals can serve as a promising ecofriendly larvicides to control mosquitoes.

This chapter summarizes the published literature regarding the antimicrobial and phytotoxic effects of plant flavonoids and discusses their potential exploitation. The authors have chosen to provide just a few examples of the diverse functionality of just one class of plant secondary metabolites the flavonoids. The chapter covers all aspects of plant flavonoid interactions with other living things in the plant environment, not strictly plant plant interactions, to cover the broadest sense of the term allelopathy. Several general reviews of flavonoid biosynthesis in plants have been published. Flavonoid biosynthesis pathways are highly regulated and controlled by both normal growth and development, as well as induction by wounding or attack by pathogens. Induced accumulation of flavonoids may be in addition to constitutive levels found in unstressed plants, which also may be transformed during the induction process.

Oxidative stress and inflammation play a key role in diverse pathological conditions such as cancer and meta-bolic disorders. The objective of this study was to determine the antioxidant and anti-inflammatory potentials of crude extract (CE) and phenolic-enriched extract (PHE) obtained from the seed coats (SCs) of black bean (BB) and pinto bean (PB) varieties. Delphinidin-3-O-glucoside (46 mg/g SC), malvidin-3-O-glucoside (29.9 mg/g SC), and petunidin-3-O-glucoside (7.5 mg/g SC) were found in major concentrations in the PHE-BB. Pelargonidin (0.53 mg/g SC) was only identified in the PHE-PB. PHE from both varieties showed antioxidant and radical scavenging capacities, with strong correlations associated with total phenolic content (TPC). Polyphenolics, including catechin, myricetin, kaempferol, quercetin, and isorhamnetin glucosides, were identified in the ex-tracts. In terms of the anti-inflammatory potentials, PHE-PB had an IC50 of 10.5 mu g dry extract/mL (mu g DE/mL) for cyclooxygenase-2 (COX-2) inhibition. The inhibition values for cyclooxygenase-1 (COX-1) ranged from 118.1 to 162.7 mu g DE/mL. Regarding inducible nitric oxide synthase (iNOS) inhibition, PHE-BB had an IC50 of 62.6 mu g DE/mL. As determined via in silico analysis, pelargonidin showed binding affinities of-7.8 and-8.5 kcal/mol for COX-1 and iNOS, respectively, and catechin had a value of-8.3 kcal/mol for COX-2. Phenolic-enriched extracts from seed coats of black and pinto beans showed good antioxidant and anti-inflammatory potential that warrants in vitro and in vivo studies.

Soybean hull biochar (SHB) was produced from soybean hulls (SH). Nine pelletized cat litter formulations at two different particle sizes (particle size 1 = between 0.84 and 2.00 mm; particle size 2 <= 0.84 mm) were produced from SHB and SH to determine water absorption (termed hydration capacity) and reduction of volatile thiols. One of these formulations from particle size 2 was chosen for additional testing. This formulation was combined with guar gum as a clumping agent and mineral oil as a dust reducer. The final formulation treated with simulated cat urine significantly reduced the population of three pathogenic bacteria over a three-day test period. Compared to four commercial biobased cat litters, this formulation was superior for odor and dust reduction while having hydration capacity and clumping ability equal to or better than the commercial litters.

This study investigated the extraction and properties of an arabinogalactan polysaccharide from frost grape (FGP) as a potential alternative to gum arabic (GA). Collection date, solvent:feed ratio (S:F), chip size, C-18 filtration, ultrafiltration, freeze drying versus spray drying, methanol pre-extraction, and water absorption were examined. Sugar composition, elemental analysis, dietary fiber content, emulsification activity index (EAI), emulsification stability index (ESI), and viscosity were used to evaluate the extracts. Exudates collected in March from live stems were viscous with high percentage solids and FGP, while May collections were watery with low percentage solids and FGP. Frost grape stems were collected, chipped, and classified by size. The extraction system utilized pressure or vacuum to increase contact between the chips and extraction water. A S:F ratio of ca. 24% gave an excellent yield of FGP. Chips between 1.8 and 3.8 mm gave the highest mass yields. Pre-extracting the chips with methanol and C-18 filtration of the water extract both yielded a lighter product. The EAI for the FGP was higher than that for GA; however, its ESI was lower. Ultrafiltration of the crude extract separated glucose, fructose, and sucrose from the FGP. FGP with glucose, fructose, and sucrose adsorbed water and became darker.

Yellow peas are a high-protein pulse crop with many potential food applications. Air-classified protein products are gaining popularity for their conserved native functionality and environmental benefits. However, air-classification of yellow peas has a few challenges preventing products from entering food markets, including fiber contamination and off-flavors/smells. A commonly recognized source of these off-flavor is due to the presence of saponins, compounds produced as secondary metabolites as a response to pathogens or environmental stress. Fungal fermentation has the potential to improve quality and broaden the applications of air-classified pea proteins in food markets. This study included a 120 h fermentation of air classified pea proteins (0.52 g/g protein) using 6 fungal organisms. Fermented material was analyzed for total phenolic content (TPC), saponin profile, mass balance, soluble and insoluble dietary fibers, and crude protein. Results indicate that fermentation can alter functional characteristics of air classified pea proteins. These alterations include potential increases in TPC, protein content, protein solubility, saponin content, and fiber fractions. Overall, these results demonstrate potential benefits of microbial fermentation processes for new and improved plant protein sources, specifically in underutilized crops like yellow peas.

Plant-based agricultural residues are readily available. However, due to the presence of several undesirable plant components such as high starch content, low protein yield, phytic acid, saponins, phenolics, etc., these feedstocks need to be processed prior to their end use1. Fermentation technology has been successful in bringing some of these feedstocks to the animal feed and human food markets by improving the nutritional composition through microbial metabolic activity2. Submerged state fermentation (SMF) is an effective way of controlling fermentation parameters (pH, temperature, agitation, etc.) to achieve high product yield and improve the quality 1,2. In this study, our goal is to use fungal fermentation to enhance the proximate composition of three different feedstocks [dry pea protein (DPP), dehulled yellow pea (DHP), and distiller's dried grains with solubles (DDGS)]. Filamentous fungi have shown varied responses with regards to cellulase production depending upon the substrate composition, leading to a change in the structural and chemical composition of the substrate3. Hence, estimation of cellulases production during submerged fermentation of different feedstocks would generate the knowledge that can be implemented to optimize the fermentation process needed for upgrading the nutritional, and economic value of the agricultural commodities The specific objectives of this study were to: 1. Ferment three substrates using three generally recognized as safe (GRAS) microbes (Aureobasidium pullulans, Neurospora crassa and Trichoderma reesei) for 120 h under submerged conditions. 2. Determine and compare the proximate compositions of substrates with their unfermented counterparts. Estimate the microbial cellulase activities at 120 h of fermentation.

BACKGROUND Canola meal has limited utilization in feed and food applications because of the presence of antinutritional factors and a high fiber content. Thus, the present study used 3-day canola seed sprouting followed by hull removal to improve the nutritional quality of canola as a feed and food ingredient to further enhance and diversify the canola market. RESULTS Seed sprouting and the hull removal process resulted in 63.2% sprouts, 29.3% mix fractions (MF) (hulls, ungerminated seed, and delayed sprouts) and 8.1% mass loss during sprouting. Fresh sprouts and MF were dried, ground and defatted to compare the obtained meals and oils with their counterparts of raw seed. Defatted sprouts (DFSP) resulted in a 46.2% reduction in crude fiber, a 34.3% reduction in acid detergent fiber and a 43.4% reduction in neutral detergent fiber compared to defatted raw seed (DFSE). DFSP provided a 10.1% higher protein content and a 5.9% increase in total amino acid content with higher essential amino acids compared to DFSE. Total carbohydrate was lowered by 5.5%, phytic acid content was lowered by 25.9%, and ash content was lowered by 5.5% in DFSP, whereas total glucosinolate content was higher in DFSP (13.1 mu mol g(-1)) than in DFSE (8.8 mu mol g(-1)). Sprouts and MF showed an oil content of 38.4% and 9.6%, respectively, compared to raw seed (34.5%). CONCLUSION Sprouting and hull removal of canola seed can potentially provide nutritive meal for food and feed applications. (c) 2022 Society of Chemical Industry.

The nutritional integrity of wheat is jeopardized by rapidly rising atmospheric carbon dioxide (CO2) and the associated emergence and enhanced virulence of plant pathogens. To evaluate how disease resistance traits may impact wheat climate resilience, 15 wheat cultivars with varying levels of resistance to Fusarium Head Blight (FHB) were grown at ambient and elevated CO2. Although all wheat cultivars had increased yield when grown at elevated CO2, the nutritional contents of FHB moderately resistant (MR) cultivars were impacted more than susceptible cultivars. At elevated CO2, the MR cultivars had more significant differences in plant growth, grain protein, starch, fructan, and macro and micro-nutrient content compared with susceptible wheat. Furthermore, changes in protein, starch, phosphorus, and magnesium content were correlated with the cultivar FHB resistance rating, with more FHB resistant cultivars having greater changes in nutrient content. This is the first report of a correlation between the degree of plant pathogen resistance and grain nutritional content loss in response to elevated CO2. Our results demonstrate the importance of identifying wheat cultivars that can maintain nutritional integrity and FHB resistance in future atmospheric CO2 conditions.

NDOLA-2 (Reg. no. GP-9, PI 698655) is an open-pollinated, non-genetically modified, spring-type canola (Brassica napus L.) developed at North Dakota State University (NDSU) and approved for specialty release by the North Dakota Agricultural Experiment Station. NDOLA-2 was tested as experimental line NDC-E16198 from 2017 to 2020 (16 location-years) in NDSU canola breeding nurseries and in 2019 and 2020 (8 location-years) in North Dakota Canola Variety Trials (ND-CVTs). NDOLA-2 has been identified as a germplasm with higher seed yield potential and disease tolerance. NDOLA-2 had 5.5% higher seed yield and 2.1% lower oil contents compared with those of commercial hybrid checks in NDSU canola breeding trials. Also, NDOLA-2 had 5.4% higher seed yield and 1.6% lower oil content over the means of commercial cultivars/hybrids in ND-CVTs. It has been identified as resistant to blackleg disease (Pathogenicity Group 4) and Sclerotinia stem rot disease of canola. NDOLA-2 flowers within 44 d from seeding, has a flowering period that lasts 22 d, and reaches maturity within 93 d from seeding with plants that are 103 cm tall. It is resistant to lodging, with a score of 2.1 out of 9, and has an overall breeder score of 8.0, where the trial mean score for all locations was 7.7.

Amaranthus hypochondriacus is a source of molecules with reported health benefits such as antioxidant activity and cancer prevention. The objective of this research was to optimize the conditions for preparing a liposome formulation using amaranth unsaponifiable matter as a source of squalene in order to minimize the particle size and to maximize the encapsulation efficiency of liposomes for carrying and delivering soybean lunasin into melanoma cell lines. Amaranth oil was extracted using supercritical dioxide carbon extraction (55.2 MPa pressure, 80 °C temperature, solvent (CO2)-to-feed (oil) ratio of 20). The extracted oil from amaranth was used to obtain the unsaponifiable enriched content of squalene, which was incorporated into liposomes. A Box–Behnken response surface methodology design was used to optimize the liposome formulation containing the unsaponifiable matter, once liposomes were optimized. Soybean lunasin was loaded into the liposomes and tested on A-375 and B16-F10 melanoma cells. The squalene concentration in the extracted oil was 36.64 ± 0.64 g/ 100 g of oil. The particle size in liposomes was between 115.8 and 163.1 nm; the squalene encapsulation efficiency ranged from 33.14% to 76.08%. The optimized liposome formulation contained 15.27 mg of phospholipids and 1.1 mg of unsaponifiable matter. Cell viability was affected by the liposome formulation with a half-maximum inhibitory concentration (IC50) equivalent to 225 μM in B16-F10 and 215 μM in A-375. The liposomes formulated with lunasin achieved 82.14 ± 3.34% lunasin encapsulation efficiency and improved efficacy by decreasing lunasin IC50 by 31.81% in B16-F10 and by 41.89% in A-375 compared with unencapsulated lunasin.

Sprouting is a beneficial way to increase the nutritional value of the original seeds. Besides, fungal fermentation of sprouts can further improve sprouts composition by reducing antinutritional factors and concentrating protein content. Thus, this study characterized the daily nutritional changes in canola sprouts and further evaluated the effect of fungal fermentation on 144 h sprouts under solid state fermentation conditions. Sprouting process resulted in high moisture containing sprouts (75.3%) due to water uptake by seeds. The oil content of sprouts (27.2% at 144 h) was significantly (p ≤ 0.05) reduced when compared to raw seeds (39.6%). Likewise, phytic acid, crude fiber, acid detergent fiber, and neutral detergent fibers were reduced by 49.7, 32.8, 19.7, and 16.6%, respectively, when compared to raw seeds. There were significant increases in protein and carbohydrate contents of sprouts, and glucosinolates also increased from 1.3 to 3.5 µM/g post sprouting. Fungal fermentation with Neurospora crassa resulted in the highest protein increase (32.8%). Heat-sterilization reduced total glucosinolates by 38.8%, and a further reduction (4.0%) was obtained by fermentation with Trichoderma reesei. A reduction in phytic acid content of 81.4, 45.8, and 10.2% was achieved by fermentation with N. crassa, T. reesei, and Aureobasidium pullulans, respectively. Total carbohydrates was reduced by 3.3 mg/mL post heat-sterilization, and fungal fermentation led to the further reduction of total carbohydrates, but total fibers were found to be increased post fermentation. These results highlight the enhancement of nutritional values of sprouted seeds and further fermented sprouts compared to ungerminated seeds and unfermented sprouts, respectively.