The primary hurdle for diagnosis of some diseases is the long incubation required to culture and confirm the presence of bacteria. The concept of using microbial VOCs as "signature markers" could provide a faster and noninvasive diagnosis. Finding biomarkers is challenging due to the specificity required in complex matrices. The objectives of this study were to (1) build/test a lab-scale platform for screening of microbial VOCs and (2) apply it to Mycobacterium avium paratuberculosis; the vaccine strain of M. bovis Bacillus Calmette-Guérin; and M. kansasii to demonstrate detection times greater those typically required for culture. SPME-GC-MS was used for sampling, sample preparation, and analyses. For objective (1), a testing platform was built for headspace sampling of bacterial cultures grown in standard culture flasks via a biosecure closed-loop circulating airflow system. For (2), results show that the suites of VOCs produced by Mycobacteria ssp. change over time and that individual strains produce different VOCs. The developed method was successful in discriminating between strains using a pooled multi-group analysis, and in timepoint-specific multi- and pair-wise comparisons. The developed testing platform can be useful for minimally invasive and biosecure collection of biomarkers associated with human, wildlife and livestock diseases for development of diagnostic point-of-care and field surveillance.

Technologies for controlling gaseous emissions of livestock is of interest to producers, the public, and regulatory agencies. In our previous lab-scale study, the use of a photocatalytic coating on surfaces subjected to black ultraviolet light reduced emissions of key odorant compounds relevant to the livestock industry. Thus, an on-farm pilot-scale experiment was conducted at a commercial swine barn to evaluate a photocatalytic coating on surfaces subjected to ultraviolet light under field conditions. A flow-through reactor was constructed with a TiO2-based photocatalytic coating on the interior surfaces and black ultraviolet light fixtures. The reactor was deployed in a room downstream of the entire swine barn exhaust. Gas samples were collected from three sampling ports in the reactor, one at the inlet (control), the midpoint (half treatment) and the outlet (treatment). Compared to the control, significant reductions in emissions were observed for p-cresol (22%), odor (16%) and nitrous oxide (9%). A significant increase in carbon dioxide (3%) was also measured. Results show that the TiO2-based photocatalytic coating and black UV light are effective in mitigating odor, a key VOC responsible for downwind swine odor, and one important greenhouse effect gas when subjected to swine barn exhaust.

In this research dataset, we summarize for the first time volatile organic compounds (VOCs) emitted in vivo from ripening wine grapes. We studied four cold-hardy cultivars grown in the Midwestern U.S.: St. Croix, Frontenac, Marquette, and La Crescent. These cultivars have gained popularity among local growers and winemakers, but still very little is known about their performance compared with long-established V. vinifera grapes. Volatiles were collected using two novel approaches: biogenic emissions from grape clusters on a vine and single grape berries. A third approach was headspace collection of volatiles from crushed grapes. Solid-phase microextraction (SPME) was used to collect volatiles. Vacuum-assisted SPME was used in the case of single grape berry. Collected VOCs were analyzed using separation and identification on a gas chromatograph mass spectrometer (GC-MS). More than 120 VOCs were identified using mass spectral libraries. The dataset provides evidence that detecting biogenic emissions from growing grapes is feasible. The dataset provides a record of temporal and spatial variability of VOCs, many of which could potentially impart aroma and flavor in the wine. The number of VOCs detected followed the order from single berry (the least) to crushed berry (the most). Thus, more information for potential use in harvesting in order to obtain the desired flavor is found in data from crushed grapes.

In this research, we propose a novel concept for a non-destructive evaluation of volatiles emitted from ripening grapes using solid-phase microextraction (SPME). This concept is novel to both the traditional vinifera grapes and the cold-hardy cultivars. Our sample models are cold-hardy varieties in the upper Midwest for which many of the basic multiyear grape flavor and wine style data is needed. Non-destructive sampling included a use of polyvinyl fluoride (PVF) chambers temporarily enclosing and concentrating volatiles emitted by a whole cluster of grapes on a vine and a modified 2 mL glass vial for a vacuum-assisted sampling of volatiles from a single grape berry. We used SPME for either sampling in the field or headspace of crushed grapes in the lab and followed with analyses on gas chromatography-mass spectrometry (GC-MS). We have shown that it is feasible to detect volatile organic compounds (VOCs) emitted in-vivo from single grape berries (39 compounds) and whole clusters (44 compounds). Over 110 VOCs were released to headspace from crushed berries. Spatial (vineyard location) and temporal variations in VOC profiles were observed for all four cultivars. However, these changes were not consistent by growing season, by location, within cultivars, or by ripening stage when analyzed by multivariate analyses such as principal component analysis (PCA) and hierarchical cluster analyses (HCA). Research into aroma compounds present in cold-hardy cultivars is essential to the continued growth of the wine industry in cold climates and diversification of agriculture in the upper Midwestern area of the U.S.

Finding farm-proven, robust sampling technologies for measurement of odorous volatile organic compounds (VOCs) and evaluating the mitigation of nuisance emissions continues to be a challenge. The objective of this research was to develop a new method for quantification of odorous VOCs in air using time-weighted average (TWA) sampling. The main goal was to transform a fragile lab-based technology (i.e., solid-phase microextraction, SPME) into a rugged sampler that can be deployed for longer periods in remote locations. The developed method addresses the need to improve conventional TWA SPME that suffers from the influence of the metallic SPME needle on the sampling process. We eliminated exposure to metallic parts and replaced them with a glass tube to facilitate diffusion from odorous air onto an exposed SPME fiber. A standard gas chromatography (GC) liner recommended for SPME injections was adopted for this purpose. Acetic acid, a common odorous VOC, was selected as a model compound to prove the concept. GC with mass spectrometry (GC⁻MS) was used for air analysis. An SPME fiber exposed inside a glass liner followed the Fick's law of diffusion model. There was a linear relationship between extraction time and mass extracted up to 12 h (R² > 0.99) and the inverse of retraction depth (1/Z) (R² > 0.99). The amount of VOC adsorbed via the TWA SPME using a GC glass liner to protect the SPME was reproducible. The limit of detection (LOD, signal-to-noise ratio (S/N) = 3) and limit of quantification (LOQ, S/N = 5) were 10 and 18 µg·m-3 (4.3 and 7.2 ppbV), respectively. There was no apparent difference relative to glass liner conditioning, offering a practical simplification for use in the field. The new method related well to field conditions when comparing it to the conventional method based on sorbent tubes. This research shows that an SPME fiber exposed inside a glass liner can be a promising, practical, simple approach for field applications to quantify odorous VOCs.

Current standard odor measurement methods arc lab-based and require substantial investment in hardware, sample collection, training, and maintenance. Odor samples must be collected in the field using bags and brought to the lab to test. This can be a time-consuming process, with the possibility of the sample loss. The actual odor measurements are based on dilution olfactometry, embodied in the AC'SCENT(R) International Olfactometer, following the ASTM E679-04 standard. In recent years a portable olfactometer, the Scentroid SM100i, has been developed for odor measurements. The portable olfactometer has many advantages over lab-based standard method, especially the lower cost-per-sample. However, very little is known about the performance and reliability of portable olfactometer where the dilutions are controlled with orifices in metallic plates. It is important to evaluate the Scentroid SM100i accuracy to determine the usefulness of using it as a comparable technology for odor measurements. The main objective of this research is to compare the performance of the lab-based ASTM E679-04 method with portable odor measurement technology. Specific objectives include: (1) determining the accuracy of the dilution ratios specified by the manufacturer of both the AC'SCENT International Olfactometer and the Scentroid SM100i;(2) comparing results between olfactometers using n-butanol, a commonly used standard gas in the olfactometry field, and (3) determining the accuracy of odor measurement using real odor samples collected from livestock farms in Iowa. The AC'SCENT olfactometer had an average percent error between the factory specifications and measured dilution ratios of 5.23% compared with 14.1% for Scentroid SM100i (using plate i-2 with dilution range most comparable to the AC'SCENT olfactometer). The use of other dilution plates resulted in average percent errors ranging from 9.68% to 25.31%. The Scentroid SM100i deviated from the manufacturer specifications for flowrates and dilution ratios, but these flowrates were generally consistent with each dilution setting. Overall, the Scentroid SM100i overestimated the odor concentrations with the mean difference of 22.9% (ranging from 0.95% to 93.34%). When the post-measurement adjustment using dilution correction was made, the mean percent average diUcrcnce was 11.8%.

Insoluble fiber (IF) recovered from the enzyme-assisted aqueous extraction process (EAEP) of soybeans is a fraction rich in carbohydrates and proteins. It can be used to enhance ethanol production in an integrated corn-soy biorefinery, which combines EAEP with traditional corn-based ethanol processing. The present study evaluated IF as a substrate for ethanol production. The effects of treatment of IF (soaking in aqueous ammonia (SAA), liquid hot water (LHW), and enzymatic hydrolysis), primarily simultaneous saccharification and co-fermentation (SSCF), as well as scaling up (250 mL to 60 L) on ethanol production from IF alone or a corn and IF slurry were investigated. Enzymatic hydrolysis (pectinase, cellulase, and xylanase, each added at 5% soy solids during simultaneous saccharification and fermentation/SSCF) was the best treatment to maximize ethanol production from IF. Ethanol yield almost doubled when SSCF of IF was performed with Saccharomyces cerevisiae and Escherichia coli KO11. Addition of IF in dry-grind corn fermentation increased the ethanol production rate (~31%), but low ethanol tolerance of E. coli KO11 was a limiting factor for employing SSCF in combination corn and IF fermentation. Nonlinear Monod modeling accurately predicted the effect of ethanol concentration on E. coli KO11 growth kinetics by Hanes-Woolf linearization. Collectively, the results from this study suggest a potential of IF as a substrate, alone or in dry-grind corn fermentation, where it enhances the ethanol production rate. IF can be incorporated in the current bioethanol industry with no added capital investment, except enzymes.

Non-invasive diagnostics and finding biomarkers of disease in humans have been a very active research area. Some of the analytical technologies used for finding biomarkers of human disease are finding their use in livestock. Non-invasive sample collection from diseased cattle using breath and headspace of fecal samples have been reported. In this work, we explore the use of volatile organic compounds (VOCs) emitted from bovine nasal secretions and serum for finding biomarkers for bovine respiratory disease (BRD). One hundred nasal swabs and 100 serum samples (n = 50 for both 'sick' and 'healthy') were collected at the time of treatment for suspected BRD. Solid-phase microextraction (SPME) was used to collect headspace samples that were analyzed using gas chromatography-mass spectrometry (GC-MS). It was possible to separate sick cattle using non-invasive analyses of nasal swabs and also serum samples by analyzing and comparing volatiles emitted from each group of samples. Four volatile compounds were found to be statistically significantly different between 'sick' and 'normal' cattle nasal swabs samples. Five volatile compounds were found to be significantly different between 'sick' and 'normal' cattle serum samples, with phenol being the common marker. Future studies are warranted to improve the extraction efficiency targeting VOCs preliminarily identified in this study. These findings bring us closer to the long-term goal of real-time, animal-side detection and separation of sick cattle.

Poultry production systems are associated with emissions of odorous volatile organic compounds (VOCs), ammonia (NH 3 ), hydrogen sulfide (H 2 S), greenhouse gases, and particulate matter. Development of mitigation technologies for these emissions is important. Previous laboratory-scale research on microbial-mineral treatment has shown to be effective for mitigation of NH 3 , H 2 S and amines emissions from poultry manure. The aim of this research was to assess the effectiveness of surface application of a microbial-mineral treatment for other important odorants, i.e., phenolics and sulfur-containing VOCs. Microbial-mineral litter additive consisting of 20% (w/w) of bacteria powder (six strains of heterotrophic bacteria) and 80% of mineral carrier (perlite-bentonite) was used at a dose of 500 g∙m -2 (per ~31 kg of manure). Samples of air were collected in two series, 4 and 7 days after application of additives. An odor profile of the poultry manure was determined using simultaneous chemical and sensory analysis. Reduction levels of VOCs determined on Day 4 was between 31% and 83% for mineral adsorbent treatment and in the range of 9% and 96% for microbial-mineral additive, depending on the analyzed compound. Reduction levels on Day 7 were considerably lower than on Day 4, suggesting that the odorous VOCs treatment efficacy is relatively short. There was no significant difference between treatments consisting of microbial-mineral additive and mineral carrier alone.

Managing the environmental impacts associated with livestock production is a challenge for farmers, public and regulatory agencies. Sustainable solutions that take into account technical and socioeconomic factors are needed. For example, the comprehensive control of odors, ammonia (NH3), hydrogen sulfide (H2S), and greenhouse gas (GHG) emissions from swine production is a critical need. Stored manure is a major source of gaseous emissions. Mitigation technologies based on bio-based products such as biochar are of interest due to the potential benefits of nutrient cycling. The objective of this study was to test non-activated (non-functionalized) biochar for the mitigation of gaseous emissions from stored manure. Specifically, this included testing the effects of: (1) time; and (2) dosage of biochar application to the swine manure surface on gaseous emissions from deep-pit storage. The biochar surface application was tested with three treatments (1.14, 2.28 and 4.57 kg·m−2 manure) over a month. Significant reductions in emissions were observed for NH3 (12.7–22.6% reduction as compared to the control). Concomitantly, significant increases in CH4 emissions (22.1–24.5%) were measured. Changes to emissions of other target gases (including CO2, N2O, H2S, dimethyl disulfide/methanethiol, dimethyl trisulfide, n-butyric-, valeric-, and isovaleric acids, p-cresol, indole, and skatole) were not statistically significant. Biochar treatment could be a promising and comparably-priced option for reducing NH3 emissions from stored swine manure.

Greenhouse gas emissions (GHGs) from swine production systems are relatively well researched with the exception of emissions from land application of manure. GHGs inventories are needed for process-based modeling and science-based regulations. Thus, the objective of this observational study was to measure GHG fluxes from land application of swine manure on a typical corn field. Assessment of GHG emissions from deep injected land-applied swine manure, fall and reapplication in the spring, on a typical US Midwestern corn-on-corn farm was completed. Static chambers were used for flux measurement along with gas analysis on a GC-FID-ECD. Measured gas concentrations were used to estimate GHGs flux using four different models: linear regression, nonlinear regression, first order linear regression and the revised Hutchinson and Mosier (HMR) model, respectively for comparisons. Cumulative flux estimates after manure application of 5.85 × 105 g•ha–1 (1 ha = 0.01 km2) of CO2, 6.60 × 101 g•ha–1 of CH4, and 3.48 × 103 g•ha–1 N2O for the fall trial and 3.11 × 106 g•ha–1 of CO2, 2.95 × 103 g•ha–1 of CH4, and 1.47 × 104 g•ha–1 N2O after the spring reapplication trial were observed. The N2O net cumulative flux represents 0.595% of nitrogen applied in swine manure for the fall trial.

The swine industry, regulatory agencies, and the public are interested in farm-tested methods for controlling gaseous emissions from swine barns. In earlier lab- and pilot-scale studies, a renewable catalyst consisting of soybean peroxidase (SBP) mixed with calcium peroxide (CaO2) was found to be effective in mitigating gaseous emissions from swine manure. Thus, a farm-scale experiment was conducted at the university's 178-pig, shallow-pit, mechanically-ventilated swine barn to evaluate SBP/CaO2 as a surficial manure pit additive under field conditions. The SBP was applied once at the beginning of the 42-day experiment at an application rate of 2.28 kg m(-2) with 4.2% CaO2 added by weight. Gas samples were collected from the primary barn exhaust fans. As compared to the control, significant reductions in gaseous emissions were observed for ammonia (NH3, 21.7%), hydrogen sulfide (H2S, 79.7%), n-butyric acid (37.2%), valeric acid (47.7%), isovaleric acid (393%), indole (31.2%), and skatole (43.5%). Emissions of dimethyl disulfide/methanethiol (DMDS/MT) increased by 30.6%. Emissions of p-cresol were reduced by 14.4% but were not statistically significant. There were no significant changes to the greenhouse gas (GHG) emissions of methane (CH4), carbon dioxide (CO2) and nitrous oxide (N2O). The total (material + labor) treatment cost was $2.62 per marketed pig, equivalent to 1.5% of the pig market price. The cost of CaO2 catalyst was similar to 60% of materials cost. The cost of soybean hulls (SBP source) was $0.60 per marketed pig, i.e., only 40% of materials cost (C) 2017 Elsevier Ltd. All rights reserved.

Comprehensive control of odors, hydrogen sulfide (H2S), ammonia (NH3) and odorous volatile organic compound (VOC) emissions associated with animal production is a critical need. Current methods utilizing wind tunnels and flux chambers for measurements of gaseous emissions from area sources such as feedlots, lagoons, pastures and cropland are often criticized for potential bias. The bias is due to the temporal isolation of the emitting surface, affecting the air velocity and temperature at the surface, forcing controlled surface-to-gas-phase mass transfer, and therefore, affecting emission measurements. That bias can limit the applicability of these methods to relative and comparative measurements. In this research, a new approach is proposed to minimize these biases with a specially constructed wind tunnel. Pilot-scale experiments were conducted to evaluate an instrumented wind tunnel capable of reproducing ambient air and soil-air mass transfer conditions in real time. Matching the inside of the wind tunnel with ambient conditions is based on simultaneous measurements of wind velocity near the emitting surface, and real-time adjustment of a wind tunnel fan to minimize velocity differences. The wind tunnel’s wind speed matching parameters were first optimized in regards to wind velocity sampling rate and wind velocity match criteria. The optimum wind tunnel operating parameters chosen resulted in a 25% absolute difference in the wind velocities generated in the wind tunnel compared to the outside wind velocities. An instrumented wind tunnel capable of reproducing ambient air and soil-air mass transfer conditions in real time could be a promising tool for measuring NH3 and VOC emissions from land-applied livestock waste.

Concentrated livestock feeding operations have become a source of odorous gas emissions that impact air quality. Comprehensive and practical technologies are needed for a sustainable mitigation of the emissions. In this study, we advance the concept of using a catalyst for barn walls and ceilings for odor mitigation. Two catalysts, a new TiO2-based catalyst, PureTi Clean, and a conventional Evonik (formerly Degussa, Evonik Industries, Essen, Germany) P25 (average particle size 25 nm) catalyst, were studied for use in reducing simulated odorous volatile organic compound (VOC) emissions on a laboratory scale. The UV source was black light. Dimethyl disulfide (DMDS), diethyl disulfide (DEDS), dimethyl trisulfide (DMTS), butyric acid, p-cresol, and guaiacol were selected as model odorants. The effects of the environmental parameters, the presence of swine dust covering the catalyst, the catalyst type and layer density, and the treatment time were tested. The performance of the PureTi catalyst at 10 µg/cm2 was comparable to that of P25 at 250 µg/cm2. The odorant reduction ranged from 100.0 ± 0.0% to 40.4 ± 24.8% at a treatment time of 200 s, simulating wintertime barn ventilation. At a treatment time of 40 s (simulating summertime barn ventilation), the reductions were lower (from 27.4 ± 8.3% to 62.2 ± 7.5%). The swine dust layer on the catalyst surface blocked 15.06 ± 5.30% of UV365 and did not have a significant impact (p > 0.23) on the catalyst performance. Significant effects of relative humidity and temperature were observed.

Bovine tuberculosis is a zoonotic disease of global public health concern. Development of diagnostic tools to improve test accuracy and efficiency in domestic livestock and enable surveillance of wildlife reservoirs would improve disease management and eradication efforts. Use of volatile organic compound analysis in breath and fecal samples is being developed and optimized as a means to detect disease in humans and animals. In this study we demonstrate that VOCs present in fecal samples can be used to discriminate between non-vaccinated and BCG-vaccinated cattle prior to and after Mycobacterium bovis challenge.

The livestock and poultry production industry, regulatory agencies, and researchers lack a current, science-based guide and data base for evaluation of air quality mitigation technologies. Data collected from science-based review of mitigation technologies using practical, stakeholders-oriented evaluation criteria to identify knowledge gaps/needs and focuses for future research efforts on technologies and areas with the greatest impact potential is presented in the Literature Database tab on the air management practices tool (AMPAT). The AMPAT is web-based (available at www.agronext.iastate.edu/ampat) and provides an objective overview of mitigation practices best suited to address odor, gaseous, and particulate matter (PM) emissions at livestock operations. The data was compiled into Excel spreadsheets from a literature review of 265 papers was performed to (1) evaluate mitigation technologies performance for emissions of odor, volatile organic compounds (VOCs), ammonia (NH3), hydrogen sulfide (H2S), particulate matter (PM), and greenhouse gases (GHGs) and to (2) inform future research needs.

Comprehensive control of odors, hydrogen sulfide (H2S), ammonia (NH3), and greenhouse gas (GHG) emissions associated with swine production is a critical need. The objective of this paper is to review the use of soybean peroxidase (SBP) and peroxides as a manure additive to mitigate emissions of odorous volatile organic compounds (VOC), NH3, H2S, and GHGs. Soybean peroxidase plus peroxide (SBPP) was tested as a mitigation technology for swine manure emissions on three scales (lab, pilot and farm). Several laboratory scale experiments were completed to assess SBPP dosages and type of oxygen source mixed into swine manure and surface application. A pilot scale experiment was done with surface application of SBPP and multiple dosages to observe scale up effects. Finally, a farm scale trial was completed to assess the SBPP treatment to a swine manure surface under a fully slatted barn floor. The ‘gated’ approach to testing SBPP from labto pilotand finally the farm-scale was appropriate and allowed for controlled experiments with sufficient replication. This approach resulted in gradual decrease of the dose of SBP, decreasing the cost of treatment, increase of treatment longevity, inclusion of many key gases of concern to the experimental protocol, and finally testing the treatment on farm-scale. To date, the farm-scale results indicate that SBPP can be effective in mitigating many important odorous gas emissions without increasing GHGs. Specifically, a 2.28 kg m-2 SBP dose mixed with 4.2% CaO2 added by weight and added to manure surface resulted in significant reductions in gaseous emissions of NH3 (21.7%), H2S (79.7%), n-butyric acid (37.2%), valeric acid (47.7%), isovaleric acid (39.3%), indole (31.2%), and skatole (43.5%). Emissions of DMDS/MT increased by 30.6%. Emissions of p-cresol were reduced by 14.4% but were not statistically significant. There were no significant changes to the GHG emissions of CH4, CO2 and N2O. The treatment cost (SBP+CaO2) was $1.45 per marketed pig of which the cost of SBP was only ~40%. Thus, further research is needed to optimize the dose and the cost of catalysts. .

The livestock and poultry production industry lacks a current, science-based guide for evaluation of air quality mitigation technologies. Therefore, we performed a science-based review of mitigation technologies using practical, stakeholders-oriented evaluation criteria to identify knowledge gaps/needs and focuses for future research efforts on technologies and areas with the greatest impact potential. Our objectives were to (1) present a recently completed Literature Database, and (2) identify and rank research needs and knowledge gaps based on the Literature Database. The Air Management Practices Tool (AMPAT) is web-based (available at www.agronext.iastate.edu/ampat) and provides an objective overview of mitigation practices best suited to address odor, gaseous, and particulate matter (PM) emissions at livestock operations. This tool helps livestock and poultry producers compare and explore different mitigation technologies. Simultaneously, a literature review of 267 papers was performed to evaluate mitigation technologies performance for emissions of odor, VOCs, NH3, H2S, PM, and GHGs and inform future research needs. Swine production systems were the most researched with 52% of the data entries. Housing and manure storage were the most researched sources of emissions with 41 and 43% of the data entries respectively. Biofilters were the most popular and farm tested technology for reducing emissions from animal housing. Aeration, anaerobic digestion, composting, diet manipulation and covers were the most researched technologies for reducing emission during manure storage and handling, with aeration being the most effective means of odor reduction farm scale tested. Injection or incorporation was the most farm tested and effective technology researched for land application.

A new method was used at the Ag 450 Farm Iowa State University (41 .98N, 93.65W) from October 24, 2012 through December 14, 2012 to assess GHG emission from land-applied swine manure on crop land. Gas samples were collected daily from four static flux chambers. Gas method detection limits were 1 .99 ppm, 17 0 ppb, and 20.7 ppb for CO2, CH4 and N2O, respectively . Measured gas concentrations were used to estimate flux using four different models, i.e., (1) linear regression, (2) non-linear regression, (3) non-equilibrium, and (4) rev ised Hutchinson & Mosier (HMR). Sixteen day s of baseline measurements (before manure application) were followed by manure application with deep injection (at 41 .2 m3/ha), and thirty seven day s of measurements after manure application. Why Study Greenhouse Gases and Land Application of Swine Manure? Assessment of greenhouse gas (GHG) emissions from land-applied swine manure is needed for improved process-based modeling of nitrogen and carbon cy cles in animal-crop production sy stems.

The economic viability of enzyme-assisted aqueous extraction processing (EAEP) of soybeans depends on properties and potential applications of all fractions (skim and insolubles as well as oil). EAEP oil contained lower free fatty acid, phosphorus, and tocopherol contents, similar unsaponifiable matter levels, and higher degrees of oxidation (peroxide and p-anisidine values) than hexane-extracted oil. The phospholipid profile of EAEP fractions was mainly composed of phosphatidic acid, followed by phosphatidylcholine, phosphatidylinositol, and phosphatidylethanolamine. Most of phospholipids were present in the skim, except for phosphatidic acid, which was the major phospholipid in the cream fraction. Skim and cream contained 55 and 3 % of the soluble carbohydrates in the original extruded flakes, respectively. Soluble carbohydrates of the skim were mainly composed of stachyose (5.8 ± 0.8 mg/mL) and sucrose (9.9 ± 0.8 mg/mL), which were hydrolyzed into glucose, galactose, and fructose after addition of α-galactosidase. Skim and cream peptides contained <20 kDa MW molecules. About 71 % of the skim peptides were <20 kDa MW, with 49 % being <1.35 kDa MW, 22 % being 17–1.35 kDa MW, and 29 % being 44–670 kDa MW. Skim protein and carbohydrate contents make this fraction suitable for replacing water in ethanol fermentations, thereby improving the fermentation rate/production and the nutritional quality of distiller’s dried grains with solubles.