Duration: 10/01/2010 to 09/30/2015

Administrative Advisor(s):

NIFA Reps:

Non-Technical Summary

Statement of Issues and Justification

The need, as indicated by stakeholders, and likely impacts from completion of the work

The most important relevant issues facing grain and livestock producers are preventing mycotoxin contamination of food and feed, and reducing the deleterious effects of mycotoxins on livestock. Mycotoxins in grains processed for biofuels becomes concentrated in the solid byproduct known as distillers grain (DG). The sale of DGs for animal feed has become an important source of supplemental income for biofuel producers, and mycotoxin contamination is a major concern. For grain buyers and food processors, the primary need is a reliable method for rapid assessment of grain quality pertaining to mycotoxins and mycotoxigenic fungi. The worst-case scenario for these stakeholders is to own millions of bushels of corn contaminated with unacceptable levels of aflatoxins and fumonisins, or of wheat with excessive concentrations of deoxynivalenol (DON). Rapid methods to detect mycotoxins at the first points of sale (elevators) as well as methods to detect mycotoxigenic fungi in the commodity (e.g. DON-producing Fusarium in barley) would help to address these concerns. Additionally, these stakeholders need cost-effective methods to predict, monitor, and minimize mycotoxin production in the field, and to detoxify mycotoxins and prevent further deterioration in contaminated grain. The lowering of tolerance limits for mycotoxins in overseas markets has increased the burden for grain buyers and food processors; currently, levels of mycotoxins that are acceptable for some US products are unacceptable in European and Asian markets, resulting in non-tariff trade barriers. New methods to monitor and treat contaminated grain would benefit domestic consumers and would allow American commodities to compete more effectively in foreign markets. Finally, workers who are responsible for animal and human health need information about the toxicity, carcinogenicity, modes of action, and biomarkers of exposure and disease for all categories of mycotoxins. This information would be used to train health-care providers to identify exposure and treat related disease, as well as to develop accurate risk assessment recommendations.

The importance of the work, and consequences if it is not done

The proposed research is wide-ranging and could lead to negative consequences if not completed. First, the presence of mycotoxins is an important health hazard. Hazard assessments including exposure assessment and evaluation of toxicity are essential. We propose basic research to define the toxicity of several important mycotoxins. Without this information, it is impossible to assess risks associated with mycotoxins. Additionally, the presence of mycotoxins in grain is an economic concern, especially in the context of global markets. Without an aggressive research program to prevent, treat, and contain outbreaks of mycotoxins in grain, US grain producers will suffer the consequences of reduced marketability of their products. Furthermore, the proposed research addresses biosecurity concerns. The natural occurrence of mycotoxins in grain is an important security concern for the grain industry and end-users of grain; mycotoxins have been used as agents of terrorism, e.g. aflatoxin in Iraq. Without a proactive research program to find innovative ways to monitor, prevent, and treat mycotoxin contamination, US agriculture will be unprepared to deal effectively with a mycotoxin outbreak, regardless of its origin. Finally, the production of mycotoxins by mycotoxigenic fungi represents a continuing problem in agriculture. Improving our understanding of how mycotoxin biosynthesis is regulated in grain-associated fungi will not only lead to novel treatment strategies, but will also advance our understanding of fungal pathogenesis in general.

The advantages for doing the work as a multistate effort and the technical feasibility of the research.

The scientists involved in this multistate, multidisciplinary research proposal work individually on mycotoxin issues related to their respective disciplines and areas of expertise. Just as agriculture is diverse and varies greatly from state to state (and in many instances, within a given state), the occurrence and severity of mycotoxin outbreaks vary widely across the US. A multistate effort ensures a thorough approach to investigate a complex and highly variable phenomenon that has significant impacts on both producers and consumers. Due to the wide range of experience and expertise of the group, the proposed research should be technically feasible.

What the likely impacts will be from successfully completing the work

The work will address the needs of the stakeholders. Outputs will include information on the action of mycotoxins in livestock and animal models. This information will be applicable to the risk assessment process. The work will also address stakeholders' continuing need for new detection and monitoring methods for grain and in DGs. Information will be generated to address the need for management practices that help prevent mycotoxin-related problems during grain production, handling, storage, processing and consumption. Finally, we will generate basic knowledge about the biochemical and molecular factors that regulate the biosynthesis of aflatoxins and Fusarium-associated mycotoxins including fumonisins and zearalenone. This will reveal critical points in the regulation where targeted controls can be developed.

Related, Current and Previous Work

NC1025 remains the only active research group that addresses the broad topic of mycotoxins, including animal health, fungal biology and genetics, analytical and detection technologies, and plant pathology. The only related Multi-state project is NC213, Management of Grain Quality and Security in World Markets, which has at least one objective that involves mycotoxin-related issues. Several members of NC1025 collaborate with members of NC213 on this research.

Over the past five years NC1025 members have completed the following activities and made the following accomplishments:

" Identified and characterized two genes responsible for the regulation of ascospore discharge in Gibberella zeae, the sexual stage of Fusarium graminearum, and a producer of deoxynivalenol.

" Identified genes encoding the polyketide synthases responsible for the biosynthesis of several mycotoxins (fusarin, aurofusarin, and zearalenone) by Fusarium graminearum.

" Identified that pyrrocidines A and B are cytotoxic to mammalian cell lines

" Identified apoptosis, initiated through both the intrinsic and the receptor mediated pathways, as the mechanism of pyrrocidine A cytotoxicity

" Modified the method for quantifying sphinganine and sphingosine in a variety of body fluids, cells and tissues to allow relatively rapid and economical confirmation of fumonisin toxicity

" Determined that selected formalin fixed tissues can be used for quantifying sphingolipid alterations to confirm fumonisin toxicity.

" Confirmed an outbreak of fumonisin induced porcine pulmonary edema (PPE) utilizing our modified method for determination of serum sphinganine and sphingosine.

" Provided evidence that DON glucuronide, the main metabolite of deoxynivalenol, was nontoxic in model human immune cells.

" Found evidence of a detoxifying adaptation of DON in blood leukocytes from mice fed the mycotoxin.

" Evaluated a number of mineral and organic adsorbents for binding mycotoxins in in vitro and in vivo studies in poultry, swine and cattle.

" Evaluated naturally occurring antioxidants (curcumin) for reducing mycotoxin toxicity in poultry.

" Produced several mycotoxins (kg quantities) for researchers doing animal feeding trials.

" Improved methods for diagnostic testing for mycotoxins in contaminated feedstuffs.

" Developed and validated real-time PCR methodology for the detection of mycotoxigenic fungi.

" Identified and characterized several genes in Fusarium verticillioides that have roles in regulation of fumonisin production and kernel colonization.

" Established that environmental moisture during grain filling is a major factor responsible for the presence of deoxynivalenol in wheat kernels with only low visual disease severity.

" Developed gas chromatography methods to analyze single wheat florets for deoxynivalenol and ergosterol, a unique fungal sterol that reflects relative biomass.

" Organized symposia with two of the MW-AOAC annual meetings, which highlighted NC1025 research.

" Maintained an Internet site that provides stakeholders with information and links about all aspects of mycotoxins.

Some of the impacts that the NC1025 activities have had include the following:

" Safe utilization of low-level mycotoxin contaminated grains in animal feedstuffs can be increased with the development of new proprietary adsorbents and use of naturally occurring antioxidants to reduce or eliminate the toxicity of these mycotoxins.

" Providing mycotoxins (fumonisin, ochratoxin A, moniliformin, zearalenone, and aflatoxin B1) in culture material to mycotoxin research groups makes it economically feasible to undertake animal feeding studies that would be nearly impossible if mycotoxins were purchased commercially.

" Characterization of genes involved in mycotoxin production and fungal development has increased our understanding of biology of the fungi and the genetic and molecular factors responsible for mycotoxin production.

" The molecular identification protocols will be applied to the analysis of distillers grain, a by-product of the ethanol production from grain that is a difficult matrix for traditional microbial techniques.

" Research that has established environmental conditions in which kernels with low disease severity nonetheless develop high levels of mycotoxin contamination has provided very useful information to those who develop models for diseases such as wheat head scab.

" Lack of deoxynivalenol glucuronide toxicity has facilitated the development of an inexpensive bioassay for DON.

" The modified method for determination of sphingolipids allows more rapid and cheaper confirmation of fumonisin toxicity.

" Demonstrated the potential toxicity of the pyrrocidines and need for its risk assessment since significant levels have been identified in unmarketable preharvest corn.

" Identified what is believed to be the first case of fumonisin toxicosis in swine (2006 corn crop) in the US since the large outbreaks of 1989.

" The committee members and their colleagues published 165 research articles between 2005 and 2009.

Objectives

1. Develop data for use in risk assessment of mycotoxins in human and animal health
   
2. Establish integrated strategies to manage and to reduce mycotoxin contamination in cereal grains and distillers grains
   
3. Define the regulation of mycotoxin biosynthesis and the molecular relationships among mycotoxigenic fungi
   

Methods

Objective 1. Develop data for use in risk assessment of mycotoxins in human and animal health

Stations participating in objective 1 (IA, MO) will continue to generate data to address the knowledge gaps related to the mechanistic basis for mycotoxin induced disease. This will be done through evaluating structure activity relationships (SAR) and investigating cell, tissue and whole animal responses at the biochemical, physiological and structural levels. Epidemiology data will be considered where available. MO will concentrate on the effects of mycotoxins on hepatic gene expression in poultry and swine. They will look at various up- and down- regulated genes that are specifically expressed in response to individual mycotoxins that will allow them to identify various pathways that control energy production and fatty acid metabolism, growth and development, antioxidant status, detoxification, coagulation, immune protection and cell proliferation. MO will continue to provide all groups with Fusarium verticillioides culture material. IA will provide Fusarium proliferatum culture material. MI, ND, IN and PA will provide trichothecene-contaminated grain samples for all stations.

1.1 Dose Response Studies and Evaluation of Potential Biomarkers

Dose response assessments will be used by IA to determine and model mycotoxin toxicity including acute toxicity, carcinogenesis, and immunomodulation. Susceptibility, such as species differences and sensitive target populations, will be considered in the dose response assessments. Exposure assessments for mycotoxins must take into account route of exposure and level and duration of exposure to the mycotoxin. Dose response data for DON and DON-glucoside in mice will be obtained by IA. Data will be generated for risk assessment of the use of mycotoxin contaminated grains, focusing on fumonisins, with both conventional and insect-resistant transgenic maize, used in the biofuels industry and its subsequent byproducts.

1.2 Structure Activity Relationships

Systematic evaluation of structure activity relationships (SAR) has not been conducted in the mycotoxin area. Fumonisin analogues have been evaluated for their cytoxicity, plant toxicity and sphingolipid alterations in only a few studies. Current activity with trichothecenes has focused on immune responses and cell signaling pathways. A Medline search revealed 14 papers in past 10 years on mycotoxin SAR. IA will use the K562 human erythroleukemia cell line as a model to evaluate SAR of DON, its major metabolites and other trichothecenes. Growth of this cell line is inhibited by DON, in a manner similar to inhibition of mouse splenocyte proliferation in vivo, according to preliminary data from Iowa. This model may be used as a biologically relevant screening assay for DON-contaminated grain and food samples; 24 h incubation of K562 cells with grain extracts containing known amounts of DON cause cell proliferation inhibition identical to the same amount of pure DON. This cell system will also be used to model human urine contents of trichothecenes, by comparing individual mycotoxin dose/responses with various combinations of trichothecenes and their key metabolites. The development of a rapid, inexpensive monitoring scheme for human exposures to these mycotoxins is a goal.

1.3 Mechanistic Studies The fumonisins have received the most attention from NC129 in the past. Rodents will be used as a model for carcinogenicity, teratogenicity and immunotoxicity for the fumonisins and DON. Since several trichothecenes have been identified by CDC as biosecurity risks, it is critical to be able to use animal assays in conjunction with cell systems to evaluate potential natural or intentional contamination of feeds and foods by these agents.

Animal assays will be critical to IA in assessing if the decontamination and detoxification strategies designed in objective 2 are successful. We will continue to assess acute toxicity of fumonisins and DON in mouse and other animal models in oral feeding studies with naturally contaminated foods or purified toxin. Adaptation to subchronic exposures to DON is a key observation in previous studies (IA) that we will assess further by studying DON metabolites and DON-metabolizing gut microbial changes over time and dose of DON exposure.

1.4 Microarrays

MO will lead an effort to explore gene expression profiles to identify candidate genes to map growth, metabolic, and regulatory pathways that control important production traits. Ribonucleic acid will be extracted from liver tissue, and its quality will be determined using gel electrophoresis and spectrophotometry. High-quality RNA will be purified from DNA contamination, reverse transcribed, and hybridized to an oligonucleotide microarray chip. Microarray data will be analyzed using a 2-step ANOVA model and validated by quantitative real-time PCR of selected genes. Genes with false discovery rates less than 13% and fold change greater than 1.4 will be considered differentially expressed. Changes in gene expression will be determined using the quantitative real-time PCR technique. Objective 2. Establish integrated strategies to monitor and reduce mycotoxin contamination in cereal grains and distillers grains

Members of the NC129/1025 have a long tradition of conducting research to test new technologies for analysis of mycotoxins and reduction of their toxic effects. The group also has collaborated in testing survey techniques. In this objective, we continue these efforts. We will use molecular techniques to develop methods to monitor mycotoxigenic fungi and test new and old technologies designed to destroy mycotoxins monitor and predict their production in the field, and prevent their toxic effects.

2.1 Surveillance

Surveillance for mycotoxins and mycotoxigenic fungi is critical for security as well as for maintaining high quality grains and grain products. Identification of fungi is expensive and time consuming; therefore, rapid and simple methods are needed to detect and monitor the presence of mycotoxigenic fungi. Information derived from these tests would allow informed decisions about processing of the product, storage life of the product, and the need for specific mycotoxin analysis.

PCR-based methods to detect mycotoxigenic fungi have provided rapid and inexpensive alternatives to techniques based on morphology. Konietzny and Greiner (2003) summarized work published between 1998 and 2003 on the use of conventional PCR methods to identify food-related Fusaria, including mycotoxigenic species. More recently, Niessen et al. (2005) and Niessesn (2007) reviewed PCR-based methods available for detection of potential producers of ochratoxins, aflatoxins, patulins, trichothecenes, and fumonisins. Although conventional PCR techniques are accurate, sensitive, and more rapid than microbiological techniques, real-time PCR techniques are even more sensitive and eliminate time-consuming techniques to analyze the reaction product, such as Southern blotting and agarose gel electrophoresis.

A comprehensive scheme for surveillance of grain and grain products to detect mycotoxigenic fungi should begin with the detection and quantification of the key mycotoxigenic genera. The information derived from the preliminary analysis would then guide subsequent analyses with mycotoxin-specific or species-specific measures. Recent Suanthie et al. (2009) developed a multiplex real-time PCR assay to detect and quantify mycotoxigenic fungi. Genus-specific Taqman probes were designed from ITS sequences of rDNA to detect Fusarium, Penicillium, and Aspergillus. The specificity of the probes was established against a wide range of fungal species. To increase the utility of assay, multiplex conditions were developed. The assay was validated by analyzing fungal growth in distillers grain (DG), an animal feedstock that accumulates as a by-product when ethanol is produced from corn.

Committee members from TX, IN, PA, ND will collaborate to expand the use of this and related DNA technologies for surveillance. One group is working to developing technology that will make this type of analysis portable. Others will continue to develop more multiplex assays from mycotoxin-specific genes. We will also work with university diagnostic labs to transfer current technology into their menu of diagnostics services. 2.2 Intervention

Grains with higher levels of contamination and most screenings from grain operations are unsafe for human and/or animal consumption and must be destroyed or alternate uses identified. If effective adsorbent clays can be identified that successfully prevent mycotoxicosis, these contaminated grains and screenings could be safely and economically utilized in the livestock and poultry industry. Committee members from MO, IA, IN, and ND will collaborate on grain research on technology that will eliminate the mycotoxins or reduce their toxicity. Research on adsorbents and natural antioxidants will be lead by the group in MO. The emphasis will be on in vitro and in vivo evaluation with respect to efficacy and to determine if they they are affective in preventing mycotoxicosis. IA will lead research that investigates chemical reactions of mycotoxins with other food constituents, with special emphasis on processes for human foods. IN and ND will evaluate methods to reduce mycotoxin contamination by the application of ozone, a highly reactive molecule, which can kill microbes and break down mycotoxins. They will explore the treatment of malting barley to reduce Fusarium mycotoxins and of corn for reduction of aflatoxin.

2.3 Fungal Population Genetics

Molecular techniques (e.g. PCR) to detect the presence of DNA of mycotoxigenic species of fungi in commodities are not only useful for quality control. They are also potentially valuable for monitoring the population genetics of fungal epidemics. This information, in turn, can help to predict and manage future epidemics. A great amount of diversity exists among field isolates of mycotoxigenic fungi in their aggressiveness and ability to produce mycotoxin, but currently there are only a few molecular markers that differentiate among these (e.g. 3ADON vs 15ADON chemotypes of Fusarium graminearum). Members from KS and KY will lead an effort to study segregation of quantitative traits involved in fecundity, aggressiveness, and mycotoxin production in F. graminearum. They will draw upon the expertise of the members from TX, PA, MI, and AR. QTL markers (AFLP markers and/or microsatellites) will be indentified for the traits. The long-term goal of this work will be to develop molecular probes that can be used to track fungal populations and allow predictions of potential mycotoxin epidemics.

Objective 3. Define the regulation of mycotoxin biosynthesis and the molecular relationships between mycotoxigenic fungi

Grains are a major source of food and energy for the worlds population. Mycotoxigenic fungi are present in essentially every agricultural field, and mycotoxin contamination represents one of the greatest continuing threats to food safety and profitability. Mycotoxin contamination of grains that enter the food, feed, and ethanol industries is estimated to cause over $100 million in losses annually. The goal of this proposal is to better understand the molecular and genetic systems that link fungal development, pathogenicity and mycotoxin production. Members of our committee have formed collaborative teams based on specific mycotoxins and fungal species.

3.1 Transcriptional Factors: Fusarium verticillioides

Transcription factors (TFs) are important regulators of fungal development, pathogenesis, and secondary metabolism (mycotoxin biosynthesis). While the genome of Fusarium verticillioides has been sequenced and 629 putative TFs classified into 41 TF families indentified, relatively few TFs have been functionally characterized. Members from TX, AR, ARS, HI, and IN will collaborate to disrupt the majority of the TF genes and characterize the resulting mutants. Generating gene knockout mutants is a highly effective experimental approach to study gene function, and we now have the molecular genetic and genomic tools to generate large numbers of mutants in a reasonable amount of time. TF knockout mutants will be evaluated for abnormalities in sexual/asexual development, secondary metabolism, and ear rot/stalk rot pathogenesis. DNA microarray analysis will be used to evaluate the genome wide effects of specific TF knockout mutants. This group is seeking grant funds to support this collaborative effort.

3.2 Sexual Reproduction: Fusarium graminearum.

F. graminearum is a common fungal pathogen of corn, wheat, and barley. Production of trichothecenes, particularly deoxynivalenol (DON = vomitoxin) by F. graminearum in wheat and barley is a major problem. Previous research has suggested that the ability of the fungus to reproduce sexually, resulting in ascospore procution, is essential for the development of field epidemics of F. graminearum. Sexual fertility in this homothallic organism is regulated by a complex mating type locus containing two different MAT genes, MAT1-1-1 and MAT1-2-1. The products of these MAT genes are TFs that regulate a cascade of other genes, many of which have unknown functions. Members from MI and KY will focus on understanding the molecular basis for sexual fertility (including production of dikaryotic hyphae and perithecia, and mechanisms of ascospore discharge). They will identify genes important to production and dissemination of ascospores in F. graminearum. They will also determine the precise roles of the individual MAT genes in these processes, and in pathogenicity to corn and wheat and toxin production, by analyzing MAT1-1-1 and MAT1-2-1 knockout mutants, and progeny of crosses between the two knockout strains. This work may identify potential antifungal targets that could be used to reduce rates of epidemic development, and thus rates of mycotoxin contamination, in the field.

3.3 Characterizing Heritable Quantitative Variation in Mycotoxin Production by F. graminearum

A majority of the North American isolates of F. graminearum belong to a single genetic lineage, known as group 7. These generally cannot be differentiated currently by molecular means, even though there is known to be a lot of diversity among different strains within the lineage. The one exception to this rule is a recently described sub-population of isolates that produces primarily 3-ADON rather than 15-ADON, and has a corresponding genetic change in the main tricothecene toxin biosynthesis gene cluster. This change can be detected by PCR. The 3-ADON chemotype currently seems to be confined mainly to Canada and the north central U.S. but it seems to be more aggressive in greenhouse studies on wheat, so there is some concern that this population may become dominant and give rise to more severe mycotoxin epidemics in the future. Use of the PCR chemotype assay will allow us to monitor the spread of the 3-ADON subpopulation in order to address this concern. Strains within the 15-ADON group are known to vary significantly in their aggressiveness and mycotoxin production, but the genetic determinants for this appear to lie outside the trichothecene toxin cluster, so that more or less mycotoxigenic and pathogenic strains cannot be differentiated with probes in this region. Currently, molecular probes do not exist that can differentiate among the dominant 15-ADON population.

In KY, studies are underway to try to understand the genetic basis for phenotypic variation in the 15-ADON group and also to map and characterize relevant QTLs. Complete genome sequences are available for two 15-ADON lineage 7 strains. These strains were crossed and random progeny are being characterized for various pathogenicity-related traits including aggressiveness to susceptible and resistant wheat varieties, and sexual and asexual fecundity. The progeny of the cross vary widely in these traits, and some are significantly more aggressive and productive than the parents, suggesting the possibility that more aggressive strains could arise in the field from crosses of less aggressive parents. Members from PA and KY will collaborate to analyze the mycotoxin profiles produced by these progeny strains and to understand the genetic basis for quantitative variation in toxin production. Understanding the molecular basis for this diversity would be of significant benefit for breeding efforts since it would allow development of more representative groups of strains for selection and screening. Furthermore, novel genes identified by QTL mapping may lead to identification of targets for new antifungal therapies.

3.4 Understanding the cause for high mycotoxin in asymptomatic grains

Fusarium species have ecological roles as pathogens, endophytes and saprotrophs and in some cases the same strain plays all 3 roles. MI and PA will work on the relationship between colonization of grains and mycotoxin contamination. Usually there is a correlation between disease symptoms and signs of the fungal pathogen on grain crops and the presence of mycotoxins but there are significant cases in which mycotoxin contamination is at levels of concern despite absent or low signs and symptoms. MI will examine the regulation of mycotoxin biosynthetic gene expression during plant colonization, in both symptomatic and asymptomatic situations and when growing as saprophytes on crop residues. PA will quantify mycotoxin levels in grains at different developmental stages and under different conditions of growth of the host plants. Results from the two approaches will be analysed and compared. Data gathered from these studies will help to clarify the relationship between fungal growth on grain crops and mycotoxin contamination.

Measurement of Progress and Results

Outputs

• Refereed journal publications; many will be co-authored by the members from multiple states.
• Development and validation of new management tools for diagnosis and prevention of mycotoxin contamination.
• Transfer of valuable research information to clientele groups (industry, government, grain producers and food producers) through general publications, website, and extension programs.

Outcomes or Projected Impacts

• The outcomes derived from the work outlined in this proposal will contribute towards the overall goals of the National Institute of Food and Agriculture (NIFA). Our results will contribute to the improvement of food security and safety, which should impact both human and animal health. U.S. and international government policy makers (e.g. USFDA, JECFA, IARC) will use our research in their risk assessments for mycotoxins. The typical outcome of risk assessment data has been government recommendations on the maximum tolerable mycotoxin levels in agricultural products, which affects both national and international use and profitability of products. We anticipate that results from the research outlined in Objective 1 will have a major impact on government decision-making by providing a better understanding of how various environmental and food processing components affect mycotoxin biosynthesis.
• Information generated from research Objective 2 will advance detection technologies that can be used by public agencies, and by private diagnostic labs to provide mycotoxin analysis services to food industries. These technologies will include development of new methodologies as well as validation of current methodologies.
• The research outlined in Objective 2 also will generate new protocols for monitoring mycotoxins in agricultural products that can be adopted by biosecurity agencies concerned with potential bioterror attacks on the nation.
• The outcomes from Objective 3 will include new basic knowledge that can be incorporated into new management strategies to help grain growers minimize mycotoxin contamination, and maximize profitability.

Milestones

(2010): We will expand the content at our website and improve the links.

(2012): We will organize a mycotoxin symposium at the Midwest AOAC annual meeting.

(2014): We will organize a mycotoxin symposium at the Midwest AOAC annual meeting.

Projected Participation

View Appendix E: Participation

Outreach Plan

The committee will continue to develop and maintain a webpage to provide information to the public. Previous website users have included news organizations, grain industry representatives, and the general public. The site incorporates contact information for members, annual reports, meeting announcements and links to all topics related to mycotoxins.

All of our outcomes derived from this research will be communicated through organized symposia. Our MW AOAC symposia have attracted researchers from states not participating in NC 1025, but have mycotoxin problems. Many of the AOAC members are involved in mycotoxin issues, and they represent industry, state and federal government. While the AOAC symposia present the work of the entire committee, each member presents his/her results at meetings specific to his/her area of expertise.

Refereed journal publications will be an important outreach tool for all the listed outcomes. Many of the publications will be on applied research. Also, members who have extension activities will transfer information to grain and food producers.

With respect to the outcomes anticipated for Objectives 2 and 3, several members will present their findings at the annual Fusarium Forum organized by the USDA Wheat and Barley Scab Initiative (www.scabusa.org). Attendees to the forum include growers, millers, representatives of industry, and scientists. The committee will collaborate with the international Fusarium Workshop (held in Kansas in odd years and at a prominent world site in even years), which provides training on biology, taxonomy, and toxicity of Fusarium species. We will coordinate with global efforts in Europe and Africa. Furthermore, the outcomes derived from Objective 1 as well as Objective 2 will be reported at the annual meetings of the Institute of Food Technologists and Experimental Biology, which are major venues for communicating on food toxicology and nutrition toxicology.

Organization/Governance

The executive committee will consist of a chair, vice-chair, secretary and past chair. The executive committee will be elected by the technical committee. Each year a new secretary will be elected and the vice-chair will advance to chair, with the chair; becoming past chair. This committee will conduct business as necessary for the whole committee, between meetings of the technical committee.

The technical committee meeting will be called once a year by the Administrative Adviser. At these meetings, work at the participating stations will be reviewed for progress and for areas needing further effort. When advantageous, efforts will be made to provide for exchange of representatives with other technical committees. Publication of results will be in the form of scientific publications, extension reports or technical bulletins, as appropriate. Attendance at the annual meeting and participation with the group will be monitored on a yearly basis. The committee will discuss with the Administrative Advisor possible remedies for delinquent members.

Duties of Members of the Executive Committee: Chair - establish location of meeting and coordinate the date with the Administrative Adviser. Notify technical committee members of dates, times and location of meeting and assist members in making accommodations. Call the meetings to order and preside during the meeting. Will become past Chair following Annual Meeting adjournment. Vice-Chair - will function as the Chair in his/her absence. Becomes chair immediately following the Annual Meeting. Is responsible for writing, getting approval and disseminating the Annual Report. Secretary - will take minutes for all meetings of the Executive Meeting and the Annual Meeting at which he/she is elected. Is responsible for disseminating copies of the minutes to all Technical Committee members following approval by the Administrative Adviser. Becomes vice chair for the next Annual Meeting.

Literature Cited

Konietzny, U., and Greiner, R., 2003. The application of PCR in the detection of mycotoxigenic fungi in foods. Brazilian Journal of Microbiology 34, 283-300.

Niessen, L., 2007. PCR-based diagnosis and quantification of mycotoxin producing fungi. International Journal of Food Microbiology 119, 38-46.

Niessen, L., Schmidt, H., Muhlencoert, E., Farber, P., Karolewiez, A., and Geisen, R., 2005. Advances in the molecular diagnosis of ochratoxin A-producing fungi. Food Additives and Contaminants 22, 324-334.

Suanthie, Y., Cousin, M.A. and Woloshuk, C. P. 2009. Multiplex real-time PCR for detection and quantification of mycotoxigenic Aspergillus, Penicillium, and Fusarium. J. Stored Prod. Res. 45:139-145.

Attachments

Land Grant Participating States/Institutions

AR, IA, IL, IN, KS, KY, MI, MO, MS, ND, NE, NJ, PA, WI

Non Land Grant Participating States/Institutions

Samuel Roberts Noble Foundation, USDA/ARS NCAUR