MARKETING AND DELIVERY OF QUALITY GRAINS AND BIOPROCESS COPRODUCTS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: TERMINATED
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 1002265
• Project No.: IND010683R
• Multistate No.: NC-_old213
• Project Start Date: Dec 30, 2013
• Project End Date: Sep 30, 2018

Project Director
Ileleji, KL, .

Recipient Organization
PURDUE UNIVERSITY
(N/A)
WEST LAFAYETTE,IN 47907

Performing Department
Ag & Biological Engineering

Non Technical Summary
In 2011 the United States harvested approximately 84 million acres of corn for grain and cash receipts from sales of corn were estimated at $63.9 billion dollars (U.S.). It also harvested 73.8 million acres of soybeans and 45.7 million acres of wheat worth an estimated $37.6 billion and $14.6 billion dollars (U.S.). NC-213 participants focus their efforts on maintaining the quality and value of these important commodities and many of the co-products produced during processing of these products. Although many of the challenges or concerns facing the industry are not new, the way in which they combine in a given harvest season is often unique. Therefore, continual adaptation and innovation are needed to maintain and improve grain and co-product quality. During the 2012 Annual Technical Meeting, a roundtable consisting of industry representatives discussed trends currently influencing the U.S. and global grain industries, and outlined research needs of the industry. These were combined with the knowledge of NC 213 researchers to give the following listing of challenges that are on the horizon. First, it is likely that the yearly volume of grain produced will continue to increase, requiring an increase in storage, handling, and drying capacity. During recent years the rise in importance of local markets (e.g. ethanol processors) and the desire on the part of producers for more marketing options have led to an increase in the volume of grain stored on farms. At the same time, public concern for food safety has increased and the Food Safety Modernization Act (FMSA) of 2012 includes requirements for supplier verification, record keeping, hazard analysis, inspection, and product tracing that will require compliance from sectors of the grain industry. Finally, additional demands are being exerted on grain supplies by new uses for grain (biofuels, bio-products). Together these challenges mean that there will be additional pressures on the industry and its management infrastructure. The opportunities and challenges presented by the above are explained in more detail in the Multistate Research Project Summary.The needs of the industry will be addressed using a collaborative approach to accomplish the following project objectives: 1) development of methods to measure grain quality and methods that preserve and improve quality; 2) development of grain storage, handling and processing systems and strategies that maintain quality, capture value and preserve food safety; and 3) effectively disseminate to the industry, research results and knowledge that can help the industry preserve and improve grain value and quality. We anticipate that the end result will be an increase in overall grain and co-product quality and purity, improvements in efficiency and effectiveness of grain handling, storage and processing facilities and systems, and increased public confidence in the quality and value of their grain based food sources.

Animal Health Component

0%

Research Effort Categories

Basic

0%

Applied

50%

Developmental

50%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
401	1510	2020	50%
712	7310	3010	10%
502	1510	2020	10%
503	1510	2020	20%
503	1510	1130	10%

Knowledge Area
502 - New and Improved Food Products; 401 - Structures, Facilities, and General Purpose Farm Supplies; 712 - Protect Food from Contamination by Pathogenic Microorganisms, Parasites, and Naturally Occurring Toxins; 503 - Quality Maintenance in Storing and Marketing Food Products;

Subject Of Investigation
7310 - Experimental design and statistical methods; 1510 - Corn;

Field Of Science
2020 - Engineering; 3010 - Economics; 1130 - Entomology and acarology;

Keywords

corn

maize

grain

grain drying

grain storage

grain handling

bioenergy

distiller's dried grains

ddgs

ozone

monitoring

insect control

mold growth

Goals / Objectives
To characterize quality and safety attributes of cereals, oilseeds, and their processed products, and to develop related measurement systems. To develop efficient operating and management systems that maintain quality, capture value, and preserve food safety in the farm-to-user supply chain. To be a multi-institutional framework for the creation of measureable impacts generated by improvements in the supply chain that maintain quality, increase value, and protect food safety/security.

Project Methods
Purdue University faculty, staff, and students will participate in NC 213 project activities, which focus on the farm-to-user supply chain that starts with raw grains and ends with processed products. World emphasis on food safety, environmental management, biosecurity, and overall quality continues to increase the need for collaboration across disciplines and institutions in developing solutions to problems and addressing the challenges faced by the farmers and industries in this chain.NC-213's annual technical meetings facilitate the interaction of professionals from USDA, universities, government agencies, and industry. It encourages informal collaborations and, through its funding of team projects, also facilitates formal collaborations among participating organizations and individuals. Purdue faculty, staff and graduate students will participate in these meetings by presenting preliminary data and will also benefit from interaction with others attendees. Purdue personnel will submit research proposals for team and individual competitions.Purdue's planned contributions to the NC-213 objectives are summarized below under each of the NC213 objectives.States that are planning related or similar research are listed using their abbreviationsObjective 1. 1a) Developing Methods to Identify, Measure, and Analyze Quality Attributes (Contributors IA, IL, IN, KS, MO, and USDA-CGAHR). 1.) Develop non-invasive imaging techniques (NIR, soft X-rays, X-ray microtomography) to determine the quality and processing characteristics of grains, oil seeds, and food products. (Contributors IA, IL, KS, IN, KY, USDA). Purdue's efforts will include evaluating the effectiveness of optical sorting for removing corn kernels infected by fungi from a corn lot thereby improving the quality of the corn lot and reducing mycotoxin levels.2.) Investigate the use of a commercially available CO2 test kit for measuring the susceptibility of shelled corn to growth of storage fungi. Purdue has been conducting tests with the CO2 test kit for almost a decade and believes the test kit has potential for improving the management of stored shelled corn by identifying corn lots that are highly susceptible to invasion by storage fungi and that therefore should be utilized before their quality deteriorates. Approaches to incorporating the test in stored corn management program will be explored and the industry will be educated on the potential use of the test to better manage stored shelled corn.3) Evaluate corn and soybean kernels to determine differences in properties between normal and diseased kernels. If resources are available, evaluate corn hybrids and soybean varieties to identify those with desirable traits that have added value to processors for specific end uses. Objective 1b) Develop Methods to Preserve and Improve Quality Attributes. (Participants IA, IN, KS, ND, OK, and USDA-CGAHR.)1.) Investigate physical and chemical processes to remove common mycotoxins (aflatoxins, DON, fumonisins, zearalenone and T-2 toxins) in dried distillers grains with solubles. Approaches include pre-cleaning and sorting and application of physical and chemical treatments. (This is an NC213 Team Project involving IN and ND.)2.) Develop methods for determining caking potential of dried distillers grains with solubles.3) Develop systems that reduce the cost of storing grain in large storage structures (i.e., large diameter steel bins, bunkers and circular ground piles) and improve the post-storage quality of grains stored in those structures. Objective 2. 2a) Improve Food Safety and Handling and Traceability of Grain and Oilseeds. (Participants IA, IL, IN, KS, MN, ND, NE, OK, and USDA-CGAHR.)1) Create and validate a survey instrument to measure quality climate (perceptions employees have about the relative importance of grain quality as compared with other business goals) within grain handling environments. Relate employee perceptions to their quality decision-making patterns in terms of food safety and security with bulk commodity crops. (Participants IA, IN).2) Develop efficient handling methods for cereal grain products and their coproducts, including: simulation models to predict flowability of grains and coproducts; simulation models to optimize reduction, mixing, and other food and feed unit operations. (Participants KS, USDA-CGAHR, IN, NE.)3) Develop harvesting and storage methods suited for developing countries that reduce postharvest corn and soybean loss during on-farm and village-level storage. (Participants IL, IN).Objective 2. 2b) Identify, Measure, and Evaluate Aflatoxin, Insects, and Other Grain Contaminants. (Participants IA, IL, IN, KS, KY, ND, NE, OK, USDA-CGAHR, and WA.)1) Investigate efficacy and economic viability of insect control technologies for food processing facilities that can be used as alternatives to current technologies such as methyl bromide fumigation and improve stored grain insect control with a 3D stored grain ecosystem and fumigant distribution model. (Participants KS, IN, MN, OK, USDA- CGAHR.) Objective 3. Fulfillment of this objective involves Multi-State Collaboration Efforts.1) Purdue NC213 participates will collaborate with other NC213 participants in conducting outreach workshops and similar activities for training grain industry personnel on the proper management of stored grain and transferring results of research to the industry.2) Purdue personnel will collaborate with other NC-213 participants in the organization of the 2015/2016 International Grain Quality and Technology Conference to be held in Manhattan KS. KS and USDA-CGAHR station representatives will be the conference co-chairs. The conference will be a forum for exchange of technical and practical information, including challenges and economic opportunities involved in creating and capturing value in the grain-based food, feed, fiber and fuel supply chains. The last International Conference was held in 2008.3) Purdue personnel will collaborate with other NC-213 participants in the development of a grain production and handling food safety- quality management system (FSQMS) template built around ISO 9000 and 22000 certification, and will adapt the FSQMS to the eastern corn belt and provide support, training, and documentation to those utilizing the system.4) Purdue personnel will collaborate with other NC-213 participants in the development of a national strategy for preserving the quality and safety of grain based food products and ensuring biosecurity within the market chain that begins with growers and ends with processed products. US grain production and market chain, with regionally or end use adapted methodologies.5) Purdue personnel will collaborate with other NC-213 participants in the training of FDA personnel and FDA-contract personnel in the conducting of food safety inspections of bulk grain and grain processing facilities under the authority of the Food Safety Modernization Act. They will also help to extend training to distance education based programs for industry professionals and assist firms with FSMA compliance in cost effective ways.6) Purdue personnel will collaborate with other NC-213 participants in the development of the International Grain Operations and Processing Center into a major industry-funded organization for distributing grain operations training and for organizing applied problem solving in emerging issues.

Progress 12/30/13 to 09/30/18

Outputs
Target Audience:Feed processing operations utilizing distillers grains with solubles. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?There were sixgraduate students and a post-doc (previously a graduate student of one of the PI). Threeof the graduate students have obtained their PhD degrees and two have obtained MS degress.Two of the former PhD students are post-docs while one is a faculty at one of the U.S. land grants. Threeof the PhD students were female and one of the females was African-American. The research has generated 8 publications (one still in review) and several of the topics were presented at conferences. How have the results been disseminated to communities of interest? What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? The four researchers listed in this Hatch project worked on four different topics of interest related to Objectives 1, 2 and 3 of the project. The topics were:?(1) Reduction in aflatoxins in corn distillers grains,?(2) Increase of shelf-life in stored wet distillers grains, (3) Evaluating the susceptibility of dried distillers grain with solubles (DDGS) to insect infestation by Tribolium castaneum, (4) Starch based hydrogel seed coating on corn germination and?(5) Characterizing dust grain dispersion and dust cloud formation.?A recap of brief highlights of the studies are given below: The effectiveness of several food additives (sodium bisulfite, sodium hypochlorite, citric acid, and ammonium persulfate) in degrading aflatoxin in samples of distillers wet grains (DWG) and condensed distillers solubles (CDS) obtained from an industrial ethanol plant was evaluated. Aflatoxin reduction was enhanced by increasing the levels of citric acid and prolonging the heating time. Another study related kernel physical characteristics to the reduction in Aflatoxin achieved using screening and density separation techniques, which are commonly used in the seed industry. The results indicate that aflatoxin levels can be reduced using a screen cleaner and a gravity table. There are currently limitations to storing and feeding distillers wet grains (DWG) due to their potential for spoilage and mycotoxin contamination in storage. A study on in-package treatments of DWG using high-voltage atmospheric cold plasma (HVACP) treatment and storage in modified atmosphere with carbon dioxide was conducted with the primary purpose of increasing product shelf-life. There was a significant difference among the various treatments to increase the shelf-life of DWG. The HVACP treatment alone and its combination with carbon dioxide-modified atmosphere storage provided a better control of microbial growth and preservation of DWG for up to 28 days of storage at 10 and 25 °C. It appears that a better strategy for the combination treatment would be to treat substrate using HVACP with modified atmospheric packaging (MAP) having a high O2 concentration (65%) and store samples post- treatment in-package in a high CO2 environment.? Germination of seeds depends on the proximity of wet soil, prolonged presence of moisture and a good soil-seed surface contact to derive water for germination and early seedling growth. This work hypothesizes that coating of seeds with a water absorbing material can help seed growth. A hydrogel is an absorbent material that can absorb and retain high quantities of water in their molecular structure, which is composed of a network of hydrophilic polymers. A hydrogel seed coating can potentially localize high quantities of water near the seed and prolong its availability. Studies subjecting seeds to unique spells of dry conditions was conducted to assess the advantages of a hydrogel coating.?While the results were promising, they were not conclusive and thus further studies on hydrogel are still needed to confirm their potential use in seed coating. Dust explosion cannot take place unless the dust cloud concentration reaches to minimum explosion limit and the severity of the explosion depend on the thermodynamics of the explosion and the kinetics, which is affected by dust dispersion, kinetic properties and environmental conditions. A dust dispersion system was developed in this study to simulate the dust leakage in the industry. Corn starch particle aerodynamic properties is being studied using this system. The effect of particles size, agglomeration degree, dust concentration in cloud and density on dust suspension and dust cloud concentration is being studied as well.

Publications

• Type: Journal Articles Status: Accepted Year Published: 2018 Citation: Fardisi, M., L.J. Mason, K.E. Ileleji and D.S. Richmond. 2018. Effect of chemical and physical properties of dried distillers grains with solubles on Tribolium castaneum (Herbst) development. Journal of Stored Product Research

Progress 10/01/16 to 09/30/17

Outputs
Target Audience:The target audience were researchers, grain storage operations and grain processing operations personnel. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?There were 3 graduate students and a post-doc (previously a graduate student of one of the PI), who researched the topics investigated as part of their PhD studies. The research generated 6 publications and several of the topics were presented at conferences. How have the results been disseminated to communities of interest?Peer-reviewed journal papers What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? The four researchers listed in this Hatch project worked on four different topics of interest related to Objectivs 1, 2 and 3 of the project. The topics were: 1. Reduction in aflatoxins in corn distillers grains. 2. Increase of shelf-life in stored wet distillers grains. 3. Evaluating the susceptibility of dried distilers grain with solubles (DDGS) to insect infestation byTribolium castaneum. 4. Starch based hydrogel seed coating on corn germination. 5. Characterizing dust grain dispersion and dust cloud formation. Brief highlights of the studies are given below: The food additives sodium bisulfite, sodium hypochlorite, citric acid, and ammonium persulfate were evaluated for their effectiveness in degrading aflatoxin in samples of distillers wet grains (DWG) and condensed distillers solubles (CDS) obtained from an industrial ethanol plant. Aflatoxin reduction was enhanced by increasing the citric acid addition level and prolonging the heating time. Unacceptably high levels of aflatoxin in shelled corn drastically reduce its market value and result in substantial economic losses. A study looked at the reduction in aflatoxin levels that could be achieved by removal of kernels discolored by mold and presumed to contain aflatoxin. The study related kernel physical characteristics to the reduction achieved using screening and density separation techniques, which are currently in use in the grain industry. The results indicate that, at least for some corn lots, aflatoxin levels can be reduced using a screen cleaner and a gravity table. There are currently limitations to storing and feeding distillers wet grains (DWG) due to their potential for spoilage and mycotoxin contamination in storage. In this study, in- package treatments of DWG using high-voltage atmospheric cold plasma (HVACP) treatment and storage in modified atmosphere with carbon dioxide were investigated with the primary purpose of increasing product shelf-life. There was a significant difference among the treatments indicated by CFU/g (P < 0.003) and pH (P < 0.0001). The HVACP treatment alone and its combination with carbon dioxide-modified atmosphere storage provided a better control of microbial growth and preservation in the wet substrate (>60% moisture), DWG for up to 28 days of storage at 10 and 25 °C. It is thought that a better strategy for the combination treatment would be to treat substrate using HVACP with modified atmospheric packaging (MAP) having a high O2 concentration (65%) and store samples post-treatment in- package in a high CO2 environment. Germination of seeds depend on the proximity of wet soil, prolonged presence of moisture and a good soil-seed surface contact to derive water for germination and early seedling growth. This work hypothesizes that coating of seeds with a water-absorbing material can help seed growth. A hydrogel is an absorbent material that can absorb and retain high quantities of water in their molecular structure, which is composed of a network of hydrophilic polymers. A hydrogel seed coating can potentially localize high quantities of water near the seed and prolong its availability. This subjected seeds to unique spells of dry conditions and assessed the advantages of a hydrogel coating. Dust explosion cannot take place unless the dust cloud concentration reaches to minimum explosion limit and the severity of the explosion depend on the thermodynamics of the explosion and the kinetics, which is affected by dust dispersion, kinetic properties and environmental conditions. A dust dispersion system was developed in this study to simulate the dust leakage in the industry. Corn starch particle aerodynamic properties will be studied using this system. The effect of particles size, agglomeration degree, dust concentration in cloud and density on dust suspension and dust cloud concentration will be studied as well.

Publications

• Type: Journal Articles Status: Published Year Published: 2017 Citation: Shi, H., R.L. Stroshine, and K.E. Ileleji. 2017. Determination of the relative effectiveness of four food additives in degrading aflatoxin in distillers wet grains and condensed distillers solubles. Journal of Food Protection 80 (1): 90-95.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Shi, H., K.E. Ileleji*, R.L. Stroshine, K.M. Keener, and J. Jensen. 2017. Reduction of aflatoxin in corn by high voltage atmospheric cold plasma. Food and Bioprocess Technology 10(6): 1042-1052.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: McClurkin-Moore, J.D., K.E. Ileleji*, and K.M. Keener. 2017. The effect of high voltage atmospheric cold plasma treatment on the shelf-life of distillers wet grains. Food and Bioprocess Technology 10(8), 1431-1440.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Shi, H., R.L. Stroshine, and K.E. Ileleji. 2017. Differences in kernel shape, size and density between healthy kernels and mold discolored kernels and their relationship to reduction in aflatoxin levels in a sample of shelled corn. Applied Engineering in Agriculture 33(3): 421-431
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Fardisi, M., L.J. Mason and K.E. Ileleji. 2017. The Susceptibility of animal feed containing dried distillers grains with solubles to Tribolium castaneum (Herbst) infestation. Journal of Stored Product Research, 72: 59-63.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Shi, H., B. Cooper, R.L. Stroshine, K.E. Ileleji and K. Keener. 2017. Structures of degradation products and degradation pathways of aflatoxin B1 by high voltage atmospheric cold plasma (HVACP) treatment. Journal of Agricultural and Food Chemistry 65(30), 6222-6230.

Progress 10/01/15 to 09/30/16

Outputs
Target Audience:Reduction of aflatoxin level in DWG and CDS by food additives The food additives sodium bisulfite, sodium hypochlorite, citric acid, and ammonium persulfate, were evaluated for their effectiveness in degrading aflatoxin in samples of distillers wet grains (DWG) and condensed distillers solubles (CDS) obtained from an industrial ethanol plant. Aqueous food additive solutions, 0.5% by weight, were added to DWG or CDS at the level of 0.5ml/g of sample, and the materials were heated at 90 °C for one hour. Sodium bisulfite was not effective in degrading aflatoxin in either DWG or CDS. Among the 4 food additives tested sodium hypochlorite was the most effective. However, it bleached the substrate and left an off-odor. Citric acid and ammonium persulfate reduced aflatoxin levels by 31-51%. Citric acid is the most promising additive for degrading aflatoxin because it has been classified as GRAS (generally recognized as safe) by FDA. Aflatoxin reduction was enhanced by increasing the citric acid addition level and prolonging the heating time. Reductions of 65% and 80% in DWG and CDS, respectively, were obtained by addition of 2.5% (by wt.) citric acid and heating at 90°C for 1 h. Aflatoxin levels in DWG and CDS were gradually reduced with prolonged heating at 90°C even without addition of food additives. Aflatoxin reductions of 53 and 73% were achieved in DWG and CDS as a result of heating at 90°C for 5 h. A number of food additives could react with AFB1, including acid, alkaline, and neutral food additives that are oxidizers. Since both DWG and CDS are acidic, a significant amount of alkaline must be added to them to treat aflatoxin. Also alkalization leaves an off-odor, making alkaline food additives a nonviable option. Among neutral food additives, sodium bisulfate, which has a relatively low oxidizing power, was not effective in aflatoxin degradation in DWG and CDS because it reacted with the substrate. The additives sodium hypochlorite and ammonium persulfate, with high oxidizing capacity, were able to significantly degrade aflatoxin in DWG and CDS. However, the distillers grains were bleached by sodium hypochlorite, which is a strong oxidizer, their nutrient value could also be affected. However, this would need to be confirmed using nutritional analyses. Citric acid appears to be the most promising food additive for degrading aflatoxin in distiller grains if there is a severe aflatoxin problem because of its efficacy and safety. During ethanol bioprocessing, the corn goes through several processes including cooking, liquefaction, fermentation, distillation, and drying of coproducts. During liquefaction and fermentation, the pH needs to be stable for maximal yield from the yeast. Therefore, the citric acid would have to be added after the fermentation process. In this study, the addition of 2.5% (by wt.) citric acid along with heating at 90°C for 1 hour reduced aflatoxin in DWG by 80% and in CDS by 61%. Addition of citric acid at a higher rate or a longer heating time would be required for higher degradation of aflatoxins. However, a balance should be achieved between aflatoxin degradation efficacy and the cost of treatment. Citric acid is 3 to 5 times more expensive than DDGS. Adding around 2.5%(by wt.) citric acid could increase the cost of distillers grains by around 10%. Reduction of aflatoxin in corn by high voltage atmospheric cold plasma The technology of high voltage atmospheric cold plasma (HVACP) generates non-thermal plasma that is widely used for microbial decontamination in medicine, material processing, and food processing. The efficacy of HVACP treatment on degradation of aflatoxin in corn was investigated. In addition, the influences of gas type (Air, MA65), relative humidity (RH 5, 40, 80%), treatment time (1, 2, 5, 10, 20 ,30 min), mode of reaction, post-treatment storage, and stirring on degradation of aflatoxin by HVACP treatment were determined. The effects of relative humidity and gas type on concentration of generated Ozone and NOx, and on optical emission spectra during HVACP treatment were also investigated. Degradation of aflatoxin B1 in corn by HVACP is mainly attributed to ozone and hydrogen peroxide generated during HVACP treatment. The majority of aflatoxin degradation during HVACP treatment occurs within minutes. The concentration of ozone and NOx increased with increasing HVACP treatment time until the ozone and NOx concentrations reached their maximum values and stabilized after10 minutes of treatment. Aflatoxin in corn was rapidly degraded by HVACP treatment. More than half of the aflatoxin was degraded within 1 minute of treatment. A greater reduction of aflatoxin could be obtained by increasing HVACP treatment time, although the increase in aflatoxin degradation in corn was less when the treatment time exceeded 10 minutes. In this study 82% of the aflatoxin in the corn was degraded after 10 minutes HVACP treatment in RH 40% air. Stirring the aflatoxin during HVACP treatment significantly increased aflatoxin degradation to 92%. The inability of the reactive species generated by HVACP to penetrate into corn kernels seemed to be the limiting factor that prevented complete degradation of aflatoxin in the corn kernels. The percent degradation of aflatoxin in corn by HVACP treatment followed a kinetic model y=a ln (b t+1) (R2adj > 0.99). MA was slightly more effective than air as a working gas for HVACP treatment for degradation of aflatoxin in corn because higher concentrations of Ozone and NOx species were generated in MA. HVACP treatment with humid air/MA at relative humidities of 40 and 80 were more effective than using dry gas (RH 5%). Direct or indrect HVACP treatments were equally effective in degrading aflatoxin. Post-treatment storage of corn with reactive gas from HVACP could slightly increase the amount of aflatoxin degraded. Results of the emission study indicated that the major emission peaks were for the nitrogen and oxygen atoms. When the air and MA emission spectra from HVACP treatment were investigated, it was found that the emission intensity of the oxygen atom decreased with increasing relative humidity and longer treatment time. The HVACP system is less efficient over a longer time period in breaking down diatomic nitrogen and oxygen. Higher concentrations of Ozone and NOx were generated during HVACP treatment in MA than in air, with lower relative humidity. Structures of Degradation Products and Degradation Pathway of AFB1 by HVACP: In this study, HVACP was applied to treat pure Aflatoxin B1 (AFB1) powder on a glass slide. HVAP treatments degraded 76% of AFB1 in 5 minutes. The Degradation products of AFB1 were separated and their formulas were elucidated using liquid chromatograph quadrupole time of flight mass spectrometers ( LC-TOF MS). Six main degradation products were observed. The structures of these degradation products were further clarified using Orbitrap Mass Spectrometry after fragmentation of the parental ions. Two degradation pathways were proposed based on the structure of degradation products. The main degradation mechanism of AFB1 involves hydrogenation, hydration, and oxidation of the furan ring. The hydrogen radical, the hydroxyl radical and ozone were proposed as the major reactive agents for AFB1 degradation generated by the HVACP treatment. According to the structure- bioactivity relationship of AFB1, the bioactivity of AFB1 sample treated by HVACP was significantly reduced due to disappearance of C8-C9 double bound in the furofuran ring in all major degradation products as well modification of lactone ring, cyclopatenone and the methoxyl group. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?We plan to demonstrate the effectiveness of cleaning on mycotoxin reduction in maize. How have the results been disseminated to communities of interest?The results have been presented at the annual NC-213 meeting and some will be written up in Extension manuscripts. What do you plan to do during the next reporting period to accomplish the goals?We plan to submit all the potential manuscripts for publiction and pursue funding to develop fundamental understanding of the HVACP process.

Impacts
What was accomplished under these goals? Reducing the levels of mycotoxins in distiller's grains will provide ethanol plants another means of controlling outbreaks of mycotoxins, and thus increase the marketability of DDGS co-products. The importance and relevance of this study were demonstrated by the 2009 and 2012 growing seasons in which a significant amount of the corn grown in Indiana (and other Midwestern states) contained mycotoxins (Deoxynevalenol in 2009 and Aflatoxin in 2012). When corn is processed into ethanol, the levels of mycotoxins in the DDGS co-product can be almost three times the levels in the incoming corn. Therefore, reduction of mycotoxin levels by removal of some of the infected kernels could significantly reduce levels of mycotoxins in the DDGS. A general indication of the effectiveness of screen cleaning, density sorting, and color sorting will allow processors to estimate the cost associated with achieving various levels of reduction. Alternative approaches involving use of solvent extraction to reduce mycotoxins in the co-product stream (objective 2) or treatments that destroy the mycotoxins present (objective 3). Results related to objective 3 indicated that some reduction can be obtained by heating at 90°C and that treatment with citric acid could be an effective means of reducing aflatoxin levels in a batch of DDGS or WDG. High voltage atmospheric cold plasma (HVACP) treatment shows promise for reducing aflatoxin and may be effective once it is commercialized and the process is better understood.

Publications

• Type: Journal Articles Status: Accepted Year Published: 2016 Citation: Shi, H., Stroshine, R.L., Ileleji, K. Determination of the Relative Effectiveness of Four Food Additives in Degrading Aflatoxin in Distillers Wet Grains and Condensed Distillers Solubles. Journal of Food Protection. 2016.
• Type: Journal Articles Status: Under Review Year Published: 2016 Citation: Shi, H., Stroshine, R.L., Ileleji, K. Differences in kernel shape, size and density between healthy and aflatoxin contaminated kernels and their relationship to reduction in aflatoxin levels in a sample of shelled corn. Applied Engineering in Agriculture. 2016.
• Type: Journal Articles Status: Under Review Year Published: 2017 Citation: Shi, H., Stroshine, R.L., Ileleji, K., Keener, K. Reduction of aflatoxin in corn by high voltage atmospheric cold plasma. Journal of Food and Bioprocess Technology. 2016,
• Type: Journal Articles Status: Under Review Year Published: 2017 Citation: Shi, H., Cooper, B., Stroshine, R.L., Ileleji, K., Keener, K. Structure of Degradation Products and Degradation Pathway of AFB1 by High Voltage Atmospheric Cold Plasma (HVACP) Treatment. Journal of Agricultural and Food Chemistry. 2016.

Progress 10/01/14 to 09/30/15

Outputs
Target Audience:Farmers, Grain Ethanol (DDGS) Processors, Grain Elevator Managers, Government Officials Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The NC-213 annual meetings provide participants with an opportunity to learn more about problems that confront the grain industry including grain processors andmanagers of grain storage facilities. The attendance of the industry advisory board members promotes interaction with industry. The informative technical presentations educate participants on a wide variety of topics of importance to the grain industry. How have the results been disseminated to communities of interest?Manuscripts are being prepared that will be submitted to professional journals. Results will also be included in presentations at extension meetings and at the NC-213 meeting. What do you plan to do during the next reporting period to accomplish the goals?The company that developed the CO2 kit is sponsoring a project for the fall of 2015 and winter of 2016 in which samples will be collected from farm bins in December, March, and, if the grain has not been sold, in late April and again in late June. These samples will be evaluated by a licensed grain inspector and also using the CO2 test kit. Results will be made available to the farmers and will eventually be published in a journal. Research on mycotoxin reduction in shelled corn will be completed. The effectiveness of HVACP treatment and of microwave heating in reducing the levels of aflatoxin in samples previously spiked with aflatoxin will be determined. The studies on fungal susceptibility testing of shelled corn and on mycotoxin reduction in DDGS will be continued during the 2015-16 reporting period. We anticipate publications on: 1) the effects of HVACP followed by storage in a high CO2 environment, 2) the effectiveness of cleaning and sorting on mycotoxin reduction in shelled corn; and 3) the effect of additives combined with heating on degradation of the aflatoxin in CDS and WDG. Studies on the effectiveness of HVACP and microwave heating on degradation of aflatoxin will be concluded. Dr. Kingsly Ambrose recently joined our research team and he will be conducting hopper flow studies on low-oil DDGS to determine flow characteristics and tendencies for segregation. In addition, he plans to conduct a study on the optimum hopper opening size for low-oil DDGS. Finally, he will study the effect of pretreatments on sorghum milling.

Impacts
What was accomplished under these goals? Purdue's current contribution to this multistate project consists of three research projects all of which align with Objective 2. The first project involves experiments on shelled corn with the Solvita (trademark) Grain CO2 test kit developed by Woods End Laboratories. Past research with the test kit indicated that it can be used to determine the fungal susceptibility (FS) of a sample of shelled corn. FS indicates how rapidly fungi will grow on the corn if the corn is exposed to conditions conducive to mold growth. If fungi grow rapidly then it is assumed the corn has a shorter "shelf life" meaning that there is greater risk of deterioration in quality of the sample caused by fungal growth. Project personnel believe the results can be used to manage stored grain to reduce the likelihood of quality deterioration caused by mold growth. After testing corn lots with higher levels of risk could be marketed first. For this first project, plans were developed for sampling and monitoring shelled corn stored on approximately 5 farms and 2 or 3 grain elevators to determine the relative fungal susceptibility of corn stored in individual bins or ground piles. More details on the CO2 test kit are included in the 2013-14 report. Samples from about 30 bins or storage piles will be gathered in the winter of 2015 and spring of 2016 and evaluated using the test kit. A second study investigated methods of increasing the time that Distillers Wet Grains (DWG) can be safely stored without quality deterioration caused by growth of fungi in the Wet Grains. This would allow more of the co-product to be utilized wet and thereby save the expense of drying. The effects of high voltage atmospheric cold plasma treatment (HVACP) combined with storage in a modified atmosphere (high CO2) were investigated as a means of extending the "shelf life" of the DWG. Results indicated that if the combination of treatments were used, the storage life of DWG could be extended to up to 28 days and up to 21 days at 10 and 25 degrees Celsius, respectively. A third study is examining methods of reducing mycotoxin levels in DDGS. This is important when the corn available for processing into ethanol contains unacceptable levels of mycotoxins. When ear rot fungi invade corn prior to harvest or when fungi grow in stored corn, they can produce mycotoxins. When contaminated corn is processed into ethanol and DDGS, the levels of mycotoxins in the DDGS can be higher than the levels in the corn that was processed. DDGS is primarily utilized as an animal feed ingredient and therefore mycotoxins make it difficult if not impossible for it to be fully utilized in feeds. Reduction of the mycotoxin levels in the DDGS could help to overcome this problem. Results of cleaning and gravity table sorting of corn prior to processing into ethanol and DDGS were reported in the 2013-14 report. This year's work focused on treatments that had the potential for transforming the mycotoxin into harmless breakdown products. Condensed soluble (CDS) and Distillers Wet Grain (DWG) were treated by heating at 60 and 90 degrees Celsius and also by conditioning the acidic materials (pH approximately 4.5) to either neutral or alkaline conditions. Heating at 90 degrees had a slight effect under acid condition, a smaller effect under neutral conditions and the greatest effect for alkaline conditions. The effect of addition of several food additives was also investigated. Additives used were those that had been reported in the literature as being effective in reducing aflatoxin levels. The samples treated with the additives were first held at 90 degrees Celsius for 1.5 hours. The most effective treatment was sodium chlorite which gave a 50% reduction. However, citric acid was considered to be the most promising because it has been classified as GRAS (generally regarded as safe). Heating for 1 hour at 90 degrees Celsius gave an 80% and 61% reduction of aflatoxin in DWG and CDS, respectively. Results of the studies are being prepared for publication. The effectiveness of HVACP and microwave treatments of aflatoxin contaminated shelled corn on reduction in aflatoxin levels are currently being studied.

Publications

Progress 12/30/13 to 09/30/14

Outputs
Target Audience: Farmers, Grain Ethanol (DDGS) Processors, Grain Elevator Managers, Government Officials. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? The NC-213 annual meetings provide participants with an opportunity to learn more about problems that confront the grain industry including grain processors andmanagers of grain storage facilities. The attendance of the industry advisory board members promotes interaction with industry. The informative technical presentations educate participants on a wide variety of topics of importance to the grain industry. How have the results been disseminated to communities of interest? Results of the research have been presented at professional meetings (ASABE, GEAPS,NC-213 Annual Meetings), and published in professional journals. They are also included in presentations at extension meetings. What do you plan to do during the next reporting period to accomplish the goals? The company that developed the CO2 kit will continue to look for people and companies involved in the grain industry who are willing to use of the CO2 kit as a tool for managing their stored corn. If time and resources permit, additional tests will be conducted to better understand how to interpret the results of the tests. Studies on invasion of DDGS by Tribolium castaneum (red flour beetle) and other insect species will be continued if support is available. Research on mycotoxin reduction in shelled corn will continue and will focus on treatment of the contaminated DDGS to reduce or eliminate the mycotoxins. Treatments planned include use of microwaves, addition of chemicals, and exposure to ozone.

Impacts
What was accomplished under these goals? Purdue’s current contribution to this multistate project consists of three research projects all of which align with Objective 2. The first project involves experiments with a CO2 test kit developed by Woods End Laboratories. When the test kit is used to evaluate shelled corn it is assumed that the results indicate how rapidly fungi will grow on the corn if it is exposed to conditions conducive to mold growth. If fungi grow rapidly then it is assumed the corn has a shorter “shelf life” meaning that there is greater risk of deterioration in quality of the sample caused by fungal growth. If the assumptions are correct then grain elevators or farmers could use the results to identify corn lots for which there is a higher risk of mold growth as a result of continued storage. Corn lots with higher levels of risk could be sold and/or processed before their quality drops, thereby decreasing losses. The second and third studies are investigating two aspects of the quality of a co-product of ethanol production, DDGS. The first of these studies is examining the susceptibility both of DDGS and of mixtures of flour and DDGS to invasion by a secondary insect feeder, tribolium castnaeum (Red Flour Beetle). If insects such as the Red Flour Beetle are going to be controlled during storage of DDGS, a better understanding is needed of the growth of this insect and other insects that invade stored grain and grain co-products. The second of the two DDGS projects has the goal of determining methods whereby mycotoxin levels in DDGS can be reduced. When ear rot fungi invade corn prior to harvest or when fungi grow in stored corn, they can produce mycotoxins. When contaminated corn is processed into ethanol and DDGS, the levels of mycotoxins in the DDGS can be higher than the levels in the corn that was processed. DDGS is primarily utilized as an animal feed ingredient and therefore mycotoxins make it difficult if not impossible for it to be fully utilized in feeds. Reduction of the mycotoxin levels in the DDGS could help to overcome this problem. For the first project, which involved the CO2 test kit, corn samples collected from rail cars by a licensed grain inspector were analyzed using the test kit. The rail cars were being shipped from elevators in west central Indiana to various destinations in the eastern and southern United States. The test kit contains a test strip with a color indicator that changes color in response to carbon dioxide in the surrounding air. The CO2 kit procedure involves first rewetting corn samples to make conditions conducive to mold growth and then incubation of the samples in sealed glass jars. When the test kit was used to evaluate the samples from the grain shipments, there were differences in CO2 production among the samples. Assuming that fungi will develop more rapidly in samples that more quickly produce CO2, the results suggest that the risk involved in continued storage of the shelled corn varied among the shipments and that there was a trend in which mold growth tended to be slightly more rapid in samples that had been stored for longer periods of time prior to shipment. Tests were also conducted on samples from the same shipment that had been stored at room temperature for 8 to 14 months and then tested using the CO2 test kit. Results of these tests indicated that the risk of mold growth in the stored corn increases with time in storage. The first study conducted on DDGS, the study of the invasion by Tribolium castaneum, revealed that “pure” DDGS is not a suitable food source for the insect. However, when the DDGS was mixed with flour and yeast, T. castaneum larvae grew as fast as when fed on a mixture of flour and yeast alone. Additionally, DDGS having large particles was less susceptible to T. castaneum than finely ground DDGS. However, T. castaneum may be able to survive and thrive in feeds containing DDGS because they can obtain sufficient nutrition other finely ground material in the feed. The second study on DDGS is investigating methods of reducing the mycotoxin levels in dried distillers DDGS. The first aspect of this study to be completed evaluated the effectiveness of removing kernels contaminated with aflatoxin from shelled corn prior to processing. Differences between the physical properties of moldy and healthy corn kernels were determined, including size, shape and density. There were statistically significant differences in the major diameters, minor diameters, sphericities, and density of moldy and healthy corn kernels. The averages for these attributes were 12.016 mm, 5.337 mm, 0.644, and 1.215 g/cm3 for healthy kernels, and 10.551 mm, 5.969 mm, 0.757, 1.147 g/cm3 for moldy kernels, respectively. Reduction in aflatoxin levels by means of screen cleaning and gravity table separation were determined by processing a (29 bu) sample of shelled corn containing high levels of aflatoxin at a commercial seed corn processing facility. The sample had unusually high levels of fine material (10%) and removing the fine materials with a screen cleaner reduced the aflatoxin levels in the retained corn kernels by 84%. Additional cleaning using a larger screen that removed small kernels reduced aflatoxin levels by 1.8% to 9.4%. Density sorting of this same sample reduced aflatoxins levels by 12.6 to 16.4% in two successive passes in which contaminated kernels were added back to the sample before the second pass. The results indicate that, for at least some samples, aflatoxin levels can be reduced significantly in at least some corn lots by removing fine material, smaller particles and lower density kernels.

Publications

• Type: Journal Articles Status: Published Year Published: 2013 Citation: Fardisi, M., Mason, L.J., Ileleji, K.E., 2013. Development and fecundity rate of Tribolium castaneum (Herbst) on distillers dried grains with solubles. Journal of Stored Product Research 52, 74-77.
• Type: Other Status: Published Year Published: 2014 Citation: Shi, H., Stroshine, R.L., and Ileleji, K.E., 2014. Aflatoxin reduction in Corn by Cleaning and Sorting. Paper No. 14-1890901, ASABE, St. Joseph, Michigan.