Source: NORTH DAKOTA STATE UNIV submitted to
MYCOTOXINS: BIOSECURITY, FOOD SAFETY AND BIOFUELS BYPRODUCTS (NC129, NC1025)
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
Reporting Frequency
Annual
Accession No.
1008833
Grant No.
(N/A)
Project No.
ND01587
Proposal No.
(N/A)
Multistate No.
NC-_old1183
Program Code
(N/A)
Project Start Date
Dec 15, 2015
Project End Date
Sep 30, 2020
Grant Year
(N/A)
Project Director
Rao, JI, .
Recipient Organization
NORTH DAKOTA STATE UNIV
(N/A)
FARGO,ND 58105
Performing Department
Plant Sciences
Non Technical Summary
Malt, used in the production of beer, is the major value added use of barley. As Fusarium Head Blight occurs in much of the small grains production region of the USA, the contamination of malt and beer with Fusarium mycotoxins is possible. The objectives of this project are to gain a better understanding of the toxins present on barley, wheat and rye, and their development in the field and during processing. This information will aid processors in producing a safer product, and also help to establish guidelines that are fair to the producer.
Animal Health Component
0%
Research Effort Categories
Basic
30%
Applied
70%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
2041550115010%
2041550200010%
2121550115010%
2121550200010%
7121550115020%
7121550200040%
Goals / Objectives
Objective 2: Establish integrated strategies to manage and reduce mycotoxin contamination in cereals and in forages. Objective 3. Better Understand the Biology and Ecology of Mycotoxigenic Fungi. Objective 1: Develop data for use in risk assessment of mycotoxins in human and animal health.
Project Methods
Craft brewing and craft malting are driving a demand for locally produced grains and malt in areas that can be severely impacted by Fusarium Head Blight (FHB). The objectives of this study are to determine the prevalence of the tricothecene mycotoxin, deoxynivalenol (DON), in small grains produced in the eastern portion of the United States, and to study the growth of Fusarium and production of DON during the malting of contaminated wheat and rye. Samples of barley, wheat and rye will be obtained from university variety trials, as well as from local grain producers and maltsters within this region. Deoxynivalenol will be determined by GC-ECD and GC-MS. The occurrence of the masked mycotoxin, or conjugated form of DON, deoxynivalenol-3-glucoside, will be determined by LC QTOF-MS. Changes in Fusarium biomass during malting well be assessed using real-time PCR.

Progress 12/15/15 to 09/30/20

Outputs
Target Audience:The target audience during the reporting period including cereal researchers, graduate students, food and malting industry. Information was delivered through laboratory research, classroom instruction, and conference. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project has provided an opportunity for two Ph.D. students and two visiting scientists to improve their fundamental knowledge of cereal, food, physicochemical and statistics sciences. How have the results been disseminated to communities of interest?The results have been published in peer review journal and presented in national conference. What do you plan to do during the next reporting period to accomplish the goals?The next step in this work will be elucidate the mode of action of antifungal efficiency and mycotoxin inhibitory activity of Fusarium graminearum isolates using different resources of clove oil (e.g., clove bud oil, clove leaf oil). It also will evaluate the antifungal activity and mycotoxin inhibitory activity of hop essential oils against Fusarium graminearum in vitro.

Impacts
What was accomplished under these goals? In this project, parameters that impact on the formation of five EO-in-water nanoemulsions and functional properties including antifungal and mycotoxin inhibitory efficacy were evaluated in vitro. In the end, the proper-designed clove oil-in-water nanoemulsions were then applied in micro-malting process by using Fusarium infected barley seeds. Results indicated that physically stable EO-in-water nanoemulsions can be fabricated by incorporating either ≥75 wt% of long chain triacylglycerol type of oil or ≥50 wt% of medium chain triacylglycerol (MCT) into EO before homogenization and homogenized under optimized processing conditions (68.95 MPa and 2 passes). Among all selected five EOs, clove oil-in-water nanoemulsions had the greatest antifungal and mycotoxin inhibitory activities. In terms of emulsifiers, the antifungal activity was mainly dominated by EO rather than emulsifier to alter mycelial and spore cell membrane integrity. At last, clove oil-in-water nanoemulsions stabilized by three different emulsifiers (Tween 80, bovine serum albumin, quillaja saponins) were selected to apply in micro-malting process according to our germinative energy test of barley seeds. All clove oil-in-water nanoemulsions had the capability to inhibit fungal growth and DON production during the micro-malting process. Among the three emulsifiers, Tween 80-stablized clove oil nanoemulsion displayed the largest reduction of mycotoxin and least flavor impact on the final malt. The overall project showed a great potential for utilization of EO-in-water nanoemulsion as antifungal agent and mycotoxin inhibitor in the malting industry.

Publications

  • Type: Theses/Dissertations Status: Accepted Year Published: 2020 Citation: Wan Jing. Fabrication and Characterization of Antifungal Essential Oil-in-Water Nanoemulsion Delivery System Targeting Fusarium graminearum in vitro and during the Malting Process
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Jing Wan, Shaobin Zhong, Paul Schwarz, Bingcan Chen, Jiajia Rao* (2020). Clove Oil-in-Water Nanoemulsion Mitigates Growth of Fusarium graminearum and Trichothecene Mycotoxin Production during the Malting of Fusarium Infected Barley. Food Chemistry, 312, 126120.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Jing Wan, Bingcan Chen, Jiajia Rao* (2020). Occurrence and Preventive Strategies to Control Mycotoxins in Cereal-Based Food. Comprehensive Reviews in Food Science and Food Safety, 19, 928953.


Progress 10/01/18 to 09/30/19

Outputs
Target Audience:The target audience during the reporting period including cereal researchers, graduate students, food and malting industry. Information was delivered through laboratory research, classroom instruction, and conference. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project has provided an opportunity for one Ph.D. student and one visiting scientist to improve their fundamental knowledge of cereal, food, physicochemical and statistics sciences. How have the results been disseminated to communities of interest?The results have been published in peer review journal and presented in national conference. What do you plan to do during the next reporting period to accomplish the goals?The next step in this work will be elucidate the mode of action of antifungal efficiency and mycotoxin inhibitory activity of Fusarium graminearum isolates using clove oil.

Impacts
What was accomplished under these goals? In this project, parameters that impact the formation of EO-in-water nanoemulsions and functional properties including antifungal and mycotoxin inhibitory efficacy were evaluated in vitro. The proper-designed EO-in-water nanoemulsions were then applied in the micro-malting process. Results indicated that physically stable EO-in-water nanoemulsions can be fabricated by incorporating either ≥75 wt% of corn oil or ≥50 wt% of medium chain triacylglycerol (MCT) into EO before homogenization and homogenized under optimized processing conditions (68.95 MPa and 2 passes). In general, the mycotoxin inhibitory efficacy of EO was enhanced considerably in the nanoemulsion form than bulk oil. Among all selected five EOs, thyme and clove oil-in-water nanoemulsions had the greatest antifungal and mycotoxin inhibitory activities. In terms of emulsifiers, the antifungal activity was mainly dominated by EO rather than the emulsifier to alter mycelial and spore cell membrane integrity. At last, clove oil-in-water nanoemulsions stabilized by three different emulsifiers (Tween 80, bovine serum albumin, and quillaja saponins) were selected to apply in micro-malting process according to our germinative energy test of barley seeds. All clove oil-in-water nanoemulsions had the capability to inhibit fungal growth and DON production during the micro-malting process. Among the three emulsifiers, the Tween 80-stablized clove oil nanoemulsion displayed the largest reduction of mycotoxin and least flavor impact on the final malt. The overall project showed a great potential for utilization of EO-in-water nanoemulsion as an antifungal agent and mycotoxin inhibitor in the food industry.

Publications

  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Jing Wan, Shaobin Zhong, Paul Schwarz, Bingcan Chen, Jiajia Rao* (2019). Enhancement of antifungal and mycotoxin inhibitory activities of food-grade thyme oil nanoemulsions with natural emulsifiers. Food Control, 106, 106709.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Dianhui Wu, Jian Lu, Shaobin Zhong, Paul Schwarz, Bingcan Chen, Jiajia Rao* (2019). Influence of nonionic and ionic surfactants on the antifungal and mycotoxin inhibitory efficacy of cinnamon oil nanoemulsions. Food & Function, 10, 2817 - 2827.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Jing Wan, Shaobin Zhong, Paul Schwarz, Bingcan Chen, Jiajia Rao*(2019). Physical properties, antifungal and mycotoxin inhibitory activities of five essential oil nanoemulsions: impact of oil compositions and processing parameters. Food chemistry, 291, 199-206.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Dianhui Wu, Jian Lu, Shaobin Zhong, Paul Schwarz, Bingcan Chen, Jiajia Rao* (2019). Effect of chitosan coatings on physical stability, antifungal and mycotoxin inhibitory activities of lecithin stabilized cinnamon oil-in-water emulsions. Food Science and Technology, 106,98-104.


Progress 10/01/17 to 09/30/18

Outputs
Target Audience:Cereal researchers, graduate students, food and malting industry. Information was delivered through laboratory research, classroom instruction, and conference. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project has provided an opportunity for two Ph.D. students and one visiting scientist to improve their fundamental knowledge of physicochemical, biochemistry and statistics. How have the results been disseminated to communities of interest?The results have been published in peer reviewed journals and presented atnational conferences. What do you plan to do during the next reporting period to accomplish the goals?The next step in this work will be to screenthe mycotoxin inhibitory activity of essential oil in water nanoemulsion in malting system. The influence of nanoemulsion composition on germination rate and tri 5 gene production in barley seed during the malting process will also be evaluated.

Impacts
What was accomplished under these goals? The antifungal activity and mycotoxin inhibitory activity of five essential oils (thyme, clove, lemongrass, cinnamon and peppermint oil) in both bulk and nanoemulsion forms were determined using two isolates of Fusarium. graminearum. The major chemical components of essential oil had a remarkable impact on long term physical stability, antifungal activity, and inhibition of mycotoxin production. With regard to inhibition of mycotoxin production, the mycotoxin inhibitory activity of essential oils was enhanced considerably in nanoemulsion form, which was attributed to greater solubility of the essential oils. It was also noticed that the same essential oils exhibited significant difference in inhibition of mycotoxin production in the two isolates of F. graminearum.

Publications

  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Jing Wan, Shaobin Zhong, Paul Schwarz, Bingcan Chen, Jiajia Rao*. Influence of oil phase composition on antifungal and mycotoxin inhibitory activity of clove oil nanoemulsions. Food & Function, 2018, 9:2872-2882.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Dianhui Wu, Jing Wan, Jian Lu, Xingguo Wang, Shaobin Zhong, Paul Schwarz, Bingcan Chen, Jiajia Rao*. Chitosan coatings on lecithin stabilized emulsions inhibit mycotoxin production by Fusarium pathogens. Food Control, 2018, 92:276-285.


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

Outputs
Target Audience:The target audience during the reporting period including cereal researchers, graduate students, food and malting industry. Information was delivered through laboratory research, classroom instruction, and conference. The findings during the reporting period will be submitted to the peer-reviewed articles in the next month. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project has provided an opportunity for a PhD student and one visiting scientist to improve their fundamental knowledge of physicochemical, biochemistry and statistics. How have the results been disseminated to communities of interest?The results have been analyzed and discussed in a PhD research proposal. What do you plan to do during the next reporting period to accomplish the goals?The next step in this work will be screened the antifungal efficiency and mycotoxin inhibitory activity of Fusarium graminearum isolates in five different essential oils. The influence of particle size, composition and interfacial properties on antifungal activity and inhibition of mycotoxin production of selected Fusarium graminearum isolates will also be evaluated.

Impacts
What was accomplished under these goals? The primary goal of this research is to apply food-grade essential oilnanoemulsions as antifungal and detoxification agents in food systems. The physically stable clove oil-in-water nanoemulsions (pH 7) were successfully made as potential antifungal delivery systems. The pure clove-oil-in water nanoemulsions were highly unstable against droplet growth due to Ostwald ripening. Ostwald ripening could be prevented by incorporating either ≥75 wt% of corn oil, or ≥ 50 wt% of medium chain triglyceride (MCT) into clove oil before homogenization. The clove oil nanoemulsions showed high stability over 30 days with mean diameters normally <150 nm. The antifungal activity of physically stable clove oil nanoemulsions were further evaluated using effective concentration (EC) and inhibitory activities towards mycotoxinproduction in two chemotypes of Fusarium graminearum isolates. The composition of oil phase, i.e., ripening inhibitor type and concentration, in clove oil-in-water nanoemulsions had a remarkable impact on the antifungal activity as well as mycotoxins production. In general, increasing the ratio of ripening inhibitors (i.e., corn oil, MCT) levels in clove oil reduced the antifungal and mycotoxininhibitory activity of nanoemulsions. This effect is also dependent on ripening inhibitor types: under the same clove oil concentration in oil phase, the addition of MCT decreased the antifungal and mycotoxins inhibitory activity of clove oil more than corn oil. For instance, clove oil containing 75 wt% corn oil showed stronger antifungal activity (EC50= 4.15~4.67 mg/g) than the same concentration of MCT (EC50=4.67 to 5.81 mg/g). Compared with bulk clove oil, our study has also shown that mycotoxins inhibitory activity of clove was significantly enhanced when encapsulated in nanoemulsions. These results have important implications for the design of essential oil based nanoemulsions as an effective antifungal and efficient detoxification delivery system in food or other industries.

Publications


    Progress 12/15/15 to 09/30/16

    Outputs
    Target Audience:The target audience includes researchers working on mycotoxins in food products and processing, as well personnel in the major multinational brewing and craft brewing industries. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Training in mycotoxin analysis and chromatography was provided to a post-doctoral research scientist How have the results been disseminated to communities of interest?Results on the development of DON and DON3G during malting have been disseminated to maltsters and brewers in the form of webinars. Results were also presented to Fusarium researchers at the International Symposium on Fusarium Head Blight. What do you plan to do during the next reporting period to accomplish the goals?The next phase of this project will investigate factors that contribute to observed differences in DON development betwwen samples. These factors include Fusarium species/chemeotypes, Fusarium vaiability, and distribution of Fusarium within the grain.

    Impacts
    What was accomplished under these goals? Beer is the most consumed alcoholic beverage in the world. In terms of food safety, mycotoxins represent the most significant problem to brewers. The primary source of mycotoxins is brewing grains, which include barley, wheat, rye, maize and rice. The Fusarium mycotoxin, deoxynivalenol (DON), is most prevalent in North America and on a worldwide basis. Past research has shown that deoxynivalenol present on malted barley can be transferred to beer at very high levels of recovery, and thus represents a food safety threat. An understanding of Fusarium and mycotoxins from field to glass is essential to all maltsters and brewers to help maintain a safe product. However, the need is perhaps even more important to the growing craft industry. Craft brewing is the fastest growing segment of the brewing industry in the United States. At present, there are approximately 4300 craft breweries, and with continued growth, they are expected to use more than 40% of all malted grain in the United States by 2020. Craft brewers use more malt in the production of their beers, more types of malted grain, and are driving a strong demand for local to regionally grown and malted grain. This is of concern, as much of this growth in local grain production is in areas that can be severely impacted by Fusarium Head Blight, and thus be contaminated with Fusarium mycotoxins. Scientific and technical resources are also limited in many of the smaller operations. The primary objectives of this project were to determine the prevalence of Fusarium mycotoxins and congeners/conjugates in brewing grains, and to study their fate during processing. Deoxynivalenol-3-glucoside (DON3G) has previous been identified as the primary DON-conjugate in barley and wheat. Past work evaluated the presence of DON and DON3G in in barley and malt. Results showed the potential for DON and DON3G levels to increase dramatically during the malting process. DON levels in barley were observed to increase by an average of 2.2-fold during malting although there was considerable variation. On the other hand levels of DON3G increased 48-fold following malting. The balance between DON and DON3G during malting is appears to be complex, with some DON being lost during processing, additional DON being formed in association with fungal growth, and then a portion of this being converted to the conjugated form via the action of barley UDP-glucosyl transferases. Conjugation is a defense or resistance response of the plant. Current work focused on the effect of grain storage on the development of DON and DON3G during the malting of wheat. Twenty hard red spring (HRS) wheat samples from the 2015 crop in North Dakota were selected to provide a range in DON content (0.02-17.92 µg/g). DON3G levels ranged from 0.02-1.23 µg/g. Micro-malting was conducted on duplicate samples after 2 and 6 months of storage. DON was determined by GC-MS and DON-3-G by LC QTOF MS. Fusarium DNA levels in wheat and malt samples were determined by Q-PCR. When malted at 2 months of storage DON increased in all an averge of 560% over the original barely, but results ranged from 113 to 1820%. The increases in DON were likely associated with the growth of Fusarium during malting, as levels of Fusarium DNA were observed to increase and average of 69% following malting. DON levels in the malts were generally too high to be utilized in brewing. Levels of DON-3-G increased by an average of 12-fold during malting with values ranging from 1.68 to 8.86 mg/g. The increase in DON-3-G is undoubtedly due to the conversion of DON by host UDP-glucosyl transferases, which have been reported to be active in wide range of germinating cereal grains. The samples were again malted following six months of storage. Analysis of variance showed that both sample and storage time had significant effects on DON levels in malts. However the sample*storage time interaction was also significant, suggesting that samples did not respond uniformly in terms of DON production. Most of the malt samples exhibited significantly lower DON and Fusarium DNA at 6 months, while only one sample exhibited a small, but significant increase. However, malt DON fell to acceptable levels for only 2 of the 12 wheat samples with DON values that were near or below the utilization/advisory limit. While, the results of this work showed a propensity for the growth of Fusarium and production of DON during malting to decline with grain storage time, 6 months may not be adequate in commercial applications. These results should be of interest to the craft malt community, given their frequent location within FHB prone areas, limited capacity for blending and storage, and need to utilize significant portions of locally produced grain.

    Publications

    • Type: Conference Papers and Presentations Status: Other Year Published: 2016 Citation: Managing DON from Field to Glass (with Dr Gary Bergstrom). Farmer Brewer Winter Weekend, Hartwick College, Oneonta, NY, January 16, 2016 Invited Presentation. Managing DON from Field to Glass (with Gary Bergstrom). Great Lakes Hop and Barley Conference, Travese City, MI. March 16. Invited Presentation. Fusarium Head Blight in Malting and Brewing: Successes and Future Challenges. The 5th International Symposium on Fusarium Head Blight (ISFHB), Florianoplis Brazil April 8, 2016. Invited Presentation