Progress 04/01/12 to 03/31/17
Outputs
Target Audience:Target audiences include academics, plant breeders, and fuel ethanol stakeholders. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Research Associate, Niki McMaster, applied techniques to quantify mycotoxins in barley, wheat, and sorghum samples using GC/MS. Nina Wilson, Ph.D. candidate,completed her Ph.D. to screen microorganisms and detoxification enzymes to destroy DON. How have the results been disseminated to communities of interest?Results from this work were presented in a poster and brief oral presentation at the USDA-USWBSI meeting in St. Louis, MO in December, 2016. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
For goal 1, DON was measured for another year for field-grown barley provided by the Griffey lab at Virginia Tech. Goals 2, 3, and 4 were completed in prior years. Microorganisms from plant and soil samples collected in Blacksburg, VA, USA, were screened by incubation in a mineral salt media containing 100 μg/mL DON and analysis by gas chromatography mass spectrometry (GC/MS). Two mixed cultures derived from soil samples consistently decreased DON levels in assays using DON as the sole carbon source. Nuclear magnetic resonance (NMR) analysis indicated that 3-keto-4-deoxynivalenol was the major by-product of DON. Using16S rRNA sequencing, these mixed cultures, including mostly members of the genera Acinetobacter, Leadbetterella, and Gemmata, were revealed. Incubation of one of these mixed cultures with wheat samples naturally contaminated with 7.1 μg/mL DON indicated nearly complete conversion of DON to the less toxic 3-epimer-DON (3-epi-DON). Our work extends previous studies that have demonstrated the potential for bioprospecting for microorganisms from the environment to remediate or modify mycotoxins for commercial applications, such as the reduction of mycotoxins in fuel ethanol co-products.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Wilson, N., McMaster, N., Gantulga, D., Soyars, C., McCormick, S., Knott, K., Senger, R., and Schmale, D. 2017. Modification of the Mycotoxin Deoxynivalenol Using Microorganisms Isolated from Environmental Samples. Toxins 9(4): 141. doi:10.3390/toxins9040141
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Progress 10/01/15 to 09/30/16
Outputs
Target Audience:Target audiences include academics, plant breeders, and fuel ethanol stakeholders. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Research Associate, Niki McMaster, applied techniques to quantify mycotoxins in barley, wheat, and sorghum samples using GC/MS. Ph.D. candidate, Nina Wilson, continued work toward her Ph.D. to screen microorganisms and detoxification enzymes to destroy DON. How have the results been disseminated to communities of interest?Results from this work were presented in a poster and brief oral presentation at the USDA-USWBSI meeting in St. Louis, MO in December 2015. What do you plan to do during the next reporting period to accomplish the goals?The project will end in March 2017, and is nearly complete. DON assessments of field-grown barley samples will continue during the next reporting period (Goal 1). Goals 2 through 4 are complete. Work during the next performance period will be focused on detoxification of mycotoxins, including the use of microorganisms and enzymes to destroy DON; a paper should be submitted soon on this work.
Impacts
What was accomplished under these goals?
Knowledge, techniques, and products resulting from our research are aimed at providing barley producers and grains, feed, food, and fuel ethanol industries with a means to diminish mycotoxin levels and losses in grain yields and quality. Unique technologies were developed and deployed to enhance ethanol production and mitigate mycotoxins in DDGS, adding economic value and assurance of product safety in barley ethanol co-products. For goal 1, DON was measured for another year for field-grown barley provided by the Griffey lab at Virginia Tech. Goals 2, 3, and 4 are complete; data were published previously for two different milling strategies to mitigate DON in barley. We are currently testing microbes and enzymes that demonstrated DON detoxification on contaminated wheat and barley samples. Our research offers a unique approach to reduce DON in these grains, particularly in the context of ethanol co-products.
Publications
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2015
Citation:
Poster and brief Presentation at the USDA-USWBSI meeting in St. Louis, MO, December 2015.
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Progress 10/01/14 to 09/30/15
Outputs
Target Audience:Target audiences include academics, plant breeders, and fuel ethanol stakeholders. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Research Associate, Niki McMaster, developed and applied techniques to quantify mycotoxins in barley and corn DDGS samples using GC/MS. Graduate student, Nina Wilson, continued work toward her Ph.D. to screen microorganisms and detoxification enzymes to destroy DON. How have the results been disseminated to communities of interest?Results from this work were presented in two posters and two short oral presentations at the USDA-USWBSI meeting in St. Louis, MN in December, 2014. Members of the Schmale Lab visited the Vireol Bioenergy Fuel Ethanol Facility in Richmond, VA in April, 2015. What do you plan to do during the next reporting period to accomplish the goals?DON assessments of field-grown barley samples will continue during the next reporting period (Goal 1). Goal 3 is complete. Goals 2 and 4 are nearly complete. Work during the next performance period will be focused on detoxification of mycotoxins during fuel ethanol production, including the use of microorganisms and enzymes to destroy DON.
Impacts
What was accomplished under these goals?
Knowledge, techniques, and products resulting from our research are aimed at providing barley producers and grains, feed, food, and fuel ethanol industries with a means to diminish mycotoxin levels and losses in grain yields and quality. Unique technologies were developed and deployed to enhance ethanol production and mitigate mycotoxins in DDGS, adding economic value and assurance of product safety in barley ethanol co-products. For goal 1, DON was measured for another year for field-grown barley provided by the Griffey lab at Virginia Tech. Goal 3 is complete; data were published during the last performance period for two different milling strategies to mitigate DON in barley. For goals 2 and 4, a collaboration was initiated with Vireol Bioenergy in Richmond, VA, and samples of corn DDGS were received for testing. Related to these goals, we screened for microbes that could grow in the presence of 100 ppm DON and found two mixed cultures and two pure cultures that consistently detoxified DON in laboratory experiments. Sequencing analysis of the pure cultures indicated that they were Pseudomonas and Achromobacter. Nuclear magnetic resonance (NMR) analysis of one of the culture byproducts indicated that DON was converted to 3-keto-DON. In a second approach, we engineered yeast strains to be sensitive to 100 ppm DON and used them to screen library fragments generated from the mixed cultures and the Pseudomonas species and cDNA enzyme sequences created by Integrated DNA Technologies. Three library fragments and two cDNA enzyme sequences were identified that allowed the yeast to grow in the presence of 100 ppm DON. In future studies, microbes and enzymes that demonstrated DON detoxification will be tested on contaminated wheat and barley samples. Our research offers a unique approach to reduce DON in these grains, particularly in the context of ethanol co-products.
Publications
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Progress 10/01/13 to 09/30/14
Outputs
Target Audience: Target audiences include academics and fuel ethanol stakeholders. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? Research Associate Niki McMaster developed and applied techniques to quantify mycotoxins in barley and corn DDGS samples using GC/MS. Graduate student Nina Wilson started on a project to screen microorganisms and detoxification enzymes to destroy DON. How have the results been disseminated to communities of interest? Results from this work have been published in peer-reviewed publications. What do you plan to do during the next reporting period to accomplish the goals? DON assessments of field-grown barley samples will continue during the next reporting period (Goal 1). Goal 3 is complete. Goals 2 and 4 are nearly complete. New approaches to mitigate DON during fuel ethanol production will be the focus of the next performance period, including the use of microrganisms and enzymes to destroy DON.
Impacts
What was accomplished under these goals?
Knowledge, techniques, and products resulting from our research are aimed at providing barley producers and grains, feed, food, and biofuel industries with a means to diminish mycotoxin levels and losses in grain yields and quality. Novel technologies were developed and deployed to enhance ethanol production and mitigate mycotoxins in DDGS, adding economic value and assurance of product safety in barley ethanol co-products. For goal 1, DON was measured for another year for field-grown barley provided by the Griffey lab at Virginia Tech. For goals 2 and 4, data were published on DON levels in corn ethanol co-products (corn DDGS) from ethanol plants across the United States. Unfortunately, DDGS lots contaminated with unacceptable levels of DON evadeddetection prior to their commercial distribution and were consequently sold as feed products. These observations underscore the need for new and improved detection and mitigation strategies for mycotoxins in DDGS. For goal 3, data were published for two different milling strategies to mitigate DON in barley. Such strategies have the potential for commercial application in mills.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2014
Citation:
Khatibi, P.A., Wilson, J., Berger, G., Brooks, W.S., McMaster, N., Griffey, C.A., Hicks, K.B., Nghiem, N.P., and Schmale, D.G. 2014. A comparison of two milling strategies to reduce the mycotoxin deoxynivalenol in barley. Journal of Agricultural and Food Chemistry. DOI: 10.1021/jf501208x
- Type:
Journal Articles
Status:
Published
Year Published:
2014
Citation:
Khatibi, P.A., McMaster, N., Musser, R., and Schmale, D.G. 2014. Survey of Mycotoxins in Corn Distillers' Dried Grains with Solubles from Seventy-Eight Ethanol Plants in Twelve States in the U.S. in 2011. Toxins 6(4): 1155-1168.
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Progress 10/01/12 to 09/30/13
Outputs
Target Audience: Target audiences include academics and fuel ethanol stakeholders. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? Research Associate Niki McMaster developed and applied new techniques to quantify mycotoxins in DDGS samples using GC/MS. Post doctoral research associate Dash Gantulga developed new techniques to assess the ability of DON detoxification enzymes to destroy DON. How have the results been disseminated to communities of interest? Results from this work have been published in peer-reviewed publications. What do you plan to do during the next reporting period to accomplish the goals? DON assessments of field-grown barley samples will continue during the next reporting period. Goal 3 is complete, and a paper should be published soon describing this work. New approaches to mitigate DON during fuel ethanol production will be explored during the next reporting period, including the use of enzymes to destroy DON.
Impacts
What was accomplished under these goals?
Knowledge, techniques, and products resulting from our research are aimed at providing barley producers and grains, feed, food, and biofuel industries with a means to diminish mycotoxin levels and losses in grain yields and quality. Novel technologies were developed and deployed to enhance ethanol production and mitigate mycotoxins in DDGS, adding economic value and assurance of product safety in barley ethanol co-products. For goal 1, DON was measured in field-grown barley provided by the Griffey lab at Virginia Tech. For goals 2 and 4, 141 DDGS samples collected from 47 ethanol plants located in 12 states were screened for mycotoxins. Twenty five percent (35/141) of the samples contained a mean of 1 to 5 ppm DON, 3% (4/141) of the samples contained a mean greater than 5 ppm but less than 10 ppm DON, and 3% (4/141) of samples contained a mean of 10 or more ppm DON. DDGS lots contaminated with unacceptable levels of DON evaded detection prior to their commercial distribution and were consequently sold as feed products. These observations underscore the need for new and improved detection and mitigation strategies for mycotoxins in DDGS. For goal 3, whole kernels from Virginian winter barley were used to evaluate the abilities of two different milling strategies (roller milling and precision milling (FitzMill®)) for their ability to remove the hull-enriched tissue from the kernel while maintaining starch levels and reducing DON levels in the endosperm-enriched tissue. Initial milling experiments demonstrated that the precision mill system was able to reduce more DON than the roller mill but with higher starch losses. The average percent DON removed from the kernel with the roller mill was 37% and the average percent DON removed from the de-hulled kernel with the precision mill was 85%.
Publications
- Type:
Journal Articles
Status:
Accepted
Year Published:
2013
Citation:
Berger, G., Green, A., Khatibi, P.A., Brooks, W.S., Rosso, L., Liu, S., Chao, S., Griffey, C.A., and Schmale, D.G. 201_. Characterization of Fusarium head blight resistance and deoxynivalenol accumulation in hulled and hulless winter barley. Plant Disease. Accepted, October 2013. First Look. http://apsjournals.apsnet.org/doi/abs/10.1094/PDIS-05-13-0479-RE
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Liu, S., Griffey, C.A., Hall, M.D., McKendry, A.L., Chen, J.L., Brooks, W.S., Brown-Guedira, G., Van Sanford, D., and Schmale, D.G. 2013. Molecular characterization of field resistance to Fusarium head blight in two US soft red winter wheat cultivars. Theoretical and Applied Genetics 126: 2485-2498. http://link.springer.com/article/10.1007%2Fs00122-013-2149-y
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Progress 10/01/11 to 09/30/12
Outputs
OUTPUTS: Fusarium Head Blight (FHB) incidence, FHB index, and Deoxynivalenol (DON) levels of Virginia hulled and hulless barley genotypes were evaluated in FHB nurseries in Virginia. Grain samples were harvested and analyzed for toxins, and a subset of these samples was processed for ethanol and DDGS. Hulls were removed from a set of hulled barley grain lots using precision dehulling and improved roller milling methods. Toxins were quantified from different fractions (untreated kernels with hulls, kernels without hulls, and hulls without kernels). A toxin detoxification gene was expressed in yeast to reduce toxin contamination in DDGS during fuel ethanol production. Toxins were measured in corn DDGS from ethanol plants throughout the U.S. PARTICIPANTS: Schmale is the PI for this Hatch project. Research Associate Niki McMaster quantified mycotoxins in samples using GC/MS, and former postdoctoral research associate Piyum Khatibi analyzed data and contributed writing to publications. COLLABORATORS AND CONTACTS. Schmale has worked closely with Dr. Carl Griffey in the CSES department and Virginia Tech to develop and test new cultivars of wheat and barley, and with Dr. Kevin Hicks with the USDA in PA for milling strategies for barley. TRAINING OR PROFESSIONAL DEVELOPMENT. Research associate Niki McMaster acquired new skills and developed new protocols on the GC/MS for mycotoxin detection and quantification. TARGET AUDIENCES: Our project will continue to increase public awareness of mycotoxin contamination in ethanol co-products used in feed and food. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
A barley-based ethanol market continues to provide potential for new opportunities and increased production of winter barley in the Eastern United States. This will not only create an important market for barley throughout the Eastern United States, it will provide valuable byproducts including carbon dioxide, fuel pellets, high protein feed ingredients for domestic animals, and enriched food products for human consumption. With the cost of feed ingredients on the rise, animal producers are contemplating lower cost options and therefore barley specifically aimed at the feed market could provide an alternative feed crop for animal producers. This also will create greater market demand for barley. Knowledge, techniques, and products resulting from our research are aimed at providing barley producers and grains, feed, food, and biofuel industries with a means to diminish mycotoxin levels and losses in grain yields and quality. Novel technologies were developed and deployed to enhance ethanol production and mitigate mycotoxins in DDGS, adding economic value and assurance of product safety in barley ethanol co-products. Our work will assist in providing long-range improvements in Virginia agriculture and food systems through a change in practice in which mycotoxin reduction in co-products becomes a priority for ethanol plants, a change in knowledge in which agricultural professionals and grain end users become aware of DON in ethanol co-products destined for domestic animal feeds and human foods, and a change in conditions in which yeast expressing mycotoxin detoxification proteins can be incorporated into standard fuel ethanol plants. Our project will also increase public awareness of mycotoxin contamination in VA in ethanol co-products used in feed and food.
Publications
- Khatibi, P.A., Berger, G., Liu, S., Brooks, W.S., Griffey, C.A., and Schmale, D.G. 2012. Resistance to Fusarium head blight and deoxynivalenol accumulation in Virginia barley. Plant Disease, DOI: 10.1094/PDIS-07-11-0551
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