Brief Summary of Minutes of Annual Meeting

NC-1025 Annual meeting minutes  20 May 2009  Turkey Run State Park, Marshall, IN The annual meeting of NC-1025 began at 8:00 a.m. on Wednesday morning May 20, 2009 in the Dogwood room of the Turkey Run Inn in Turkey Run State Park, Marshall, Indiana. Registra-tion of $30 per person was collected for the meeting. Minutes for the previous years meeting with the Midwest AOAC in Bozeman, Montana were not submitted by Dr. David Kendra and were unavailable for review. Dr. Beverly Durgan, the projects administrative advisor, began the meeting with a reminder that the project was in its fourth year and that a renewal proposal was due by September 1. She out-lined some of the things that needed to be decided  Does the committee want to apply for a re-newal? Does the committee want to keep the current NC number and name? She also requested that we review the current national priority areas and make sure that the new/rewritten project fits within those priority areas. Since the project underwent a major rewrite the last time, she thought that at least some of the current objectives and work plans would carry over to a renewal. Project reports were made by: George Rottinghaus  University of Missouri David LeDoux  University of Missouri Frances Trail  Michigan State University Pat Murphy  Iowa State University Suzanne Hendricks  Iowa State University Gretchen Kuldau  The Pennsylvania State University Wanda Haschek  University of Illinois Champaign-Urbana John Leslie  Kansas State University Charles Woloshuk  Purdue University Steve Hooser  Purdue University Lisa Vaillancourt  University of Kentucky A break for lunch was taken after Dr. Woloshuks project report. Following the project reports, the business meeting was held followed by a discussion of the re-newal of the project. Prior to the discussion, Dr. Haschek (IUCU) announced that she was semi-retired and that a replacement for her should be found. The current years (2008) annual report is being prepared by Dr. Charlene Wolf-Hall (NDSU). The current vice-chair Dr. David Kendra (NCAUR) was not present, which resulted in a discus-sion regarding the site for the meeting in 2010. Dr. Woloshuk indicated that he received no re-sponse from Dr. Kendra when queried about hosting the meeting in Peoria next year. Dr. Fran-ces Trail (MSU) volunteered to serve as chair in 2010 and to assume the responsibility for orga-nizing next years meeting. The meeting will be in Michigan at a date and location to be decided later. Either Dr. John Leslie or Dr. J. Scott Smith of KSU will serve as vice-chair of the 2010 meeting and the chair and organizer of the 2011 meeting, assuming that the project is renewed for an additional five-year term. Dr. Woloshuk will lead the effort to prepare the termination re-port for the current five-year project when it is due next year. He will be requesting lists of rele-vant publications and interactions from all project participants in the near future. A lengthy discussion ensued that centered on the revision of the current proposal for resubmis-sion and the renewal. To be successful the group needs to be composed of an interdisciplinary mix of chemists, food scientists, plant pathologists and toxicologists. No one university has staff in all of these areas. Integrating these efforts to keep everyone current in the related fields that are important to this committee and the field of mycotoxicology was viewed as one of the most important roles filled by the committee. USDA-ARS involvement, especially from NCAUR in Peoria, Illinois, was viewed as essential for the long-term success of the project. The current proposal has four objectives. All of them will be rewritten for the new proposal, with some material deleted, some refocused and some new material added. Dr. Rottinghaus agreed to be the lead for the current Objective 1, Dr. Woloshuk for the current objective 2, and Dr. Trail for the current objective 4. Current objective 3 may not survive in its current form, with the lead to rewrite it to be determined at a later date. The section on surveys contained in current Objective 3 will be deleted. This section may be replaced with work on dry distiller grains (DDGs) from commercial ethanol fermentations. Those writing individual pieces are to have them to Dr. Woloshuk by the end of July. Potential outcomes, outputs, milestones and the lead to rewrite objective 3, if necessary, also are to be identified at this time. Two decisions were made on the general thrust of the renewal. First, the current web site will be kept and updated. Second, occasional symposia at the Midwest AOAC meetings will continue to be scheduled concurrently in a joint meeting of the committee with this group. The meeting adjourned at 3:30 p.m. Meeting attendees: Beverly Durgan (Minnesota) Wanda Haschek (Illinois) Steve Hooser (Purdue) Suzanne Hendricks (ISU) Gretchen Kuldau (Penn. State) David LeDoux (Missouri) John Leslie (KSU) Pat Murphy (ISU) George Rottinghaus (Missouri) Frances Trail (MSU) Lisa Vaillancourt (Kentucky) Charles Woloshuk (Purdue)

Objective 1. Develop data for use in risk assessment of mycotoxins in human and animal health The Illinois group set up a subcontract with Purdue University to perform additional in vivo studies investigating the effects of pyrrocidine which was previously shown to be cytotoxic in mammalian cell lines. Illinois will be providing research material to Purdue University for testing of new analytical procedures for diagnosis of fumonisin toxicity. The Iowa group hypothesized that deoxynivalenol (DON) changed leukocyte subset numbers and their migration potential in peripheral blood, with interactions of age and sex. These leukocyte markers could be functional biomarkers of DON exposure in humans. In BALB/c mice fed DON at 0, 1.0 and 2.0 ppm, age, sex and feeding interval altered immune response of peripheral blood (PB) and splenic leukocytes, as measured by flow cytometry by staining cell surface markers. In 2.5-month old female mice, a decreased percentage of PB CD4+ cells and a decreased percentage of CD11b+ (macrophage) splenic leukocytes at 2.0 ppm DON were seen after 14 d only, whereas 2.0 ppm DON caused an increased percentage of CD19+ cells after 14 and 28 d. In 16.5-month old females, decreased PB CXCR5+ B cells were noted after feeding 2.0 ppm DON for 14 d, and 1.0 ppm but not 2.0 ppm DON decreased splenic CD11b+ cell %. In old male mice, 1.0 and 2.0 ppm DON increased granulocytes and CD29+CD11a+ neutrophils after 14 d and 2.0 ppm DON increased CCR9+ T cytotoxic cells after 28 d. DONcaused no changes to immune cell markers in young male mice. Objective 2. Develop new techniques and improve current assays to identify and measure mycotoxins and mycotoxigenic fungi in cereal grains A simple method for identifying heterozygous chromosome rearrangements in Fusarium spp. was adapted by the Kansas group from techniques proven to work with Gibberella zeae. Preliminary studies with this technique sufficed to identify the chromosomal rearrangements known in the mapping cross of G. zeae and to suggest that the lineages of G. zeae are not genetically isolated by chromosome rearrangements. The peptide sex pheromones of G. zeae have been identified and the genes encoding both the peptide pheromones and their receptors cloned. Only one of the pheromone/receptor pairs appears functional in G. zeae based on the fertility and interaction patterns of various deletion mutants. The one active pheromone is fungistatic and capable of blocking germination of both conidia and ascospores. Strains in which the receptor or the pheromone are deleted are still fertile as homothallics, but are much less fertile, in terms of both perithecia and ascospores produced, than are strains in which both the pheromone and its receptor are functional. The Kansas group reported that Fusarium fujikuroi is a close relative of Fusarium proliferatum capable of producing high quantities of gibberellic acid and causing the bakane disease of rice. F. fujikuroi has recently been introduced to California, where it is of great concern. The fungus can pathogenize native grass species that grow in and near the rice fields, which should make eradication of the disease from infested fields all but impossible. The California populations were clonal, with a single genotype widely dispersed across the state. Both mating types were present in the field populations but with a huge excess of MAT-1 alleles. Both AFLP and VCG strain typing gave similar results in terms of evaluating the genetic diversity present. The Kansas group reported that two new Fusarium species one from sorghum and maize and another from finger millet have been identified. Sexual stages have been identified for both species, and tester isolates developed to text cross-fertility are available in one species but not the other. The mycotoxin production capabilities of these species are not yet known The North Dakota group reported that methods were developed for the detection and quantification of Fusarium metabolites in matrices such as barley, wheat and potatoes. Covalently bound deoxynivalenol can be detected and quantified in barley with a new GC method, and in wheat with a new HPLC method, which also detects and measures pigmented metabolites of Fusarium graminearum. The Purdue group reported that the purpose of our research was to develop a multiplex real-time PCR assay to detect and quantify mycotoxigenic fungi. Our strategy was to utilize broad-spectrum primers to amplify the targeted genomic DNA and specific fluorescent probes to detect and quantify the different fungal genera. An analysis of PCR primer indicated PCR products from more than 40 Aspergillus species, 23 Fusarium species, and 32 Penicillium species as well as 64 other fungal genera. Genus-specific Taqman probes were designed from ITS sequences of rDNA to detect mycotoxigenic 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, an animal feedstock that accumulates as a by-product when ethanol is produced from corn. Objective 3. Establish integrated strategies to manage and to prevent mycotoxin contamination in cereal grains The Fusarium/ Poultry Research Laboratory in Missouri evaluated a number of mineral and organic adsorbents for binding mycotoxins in in vitro and in vivo studies in poultry, swine and cattle. Naturally occurring antioxidants (curcumin) were evaluated for reducing mycotoxin toxicity in poultry. The laboratory continues to produce mycotoxins in culture (kg quantities) for in house as well as for other researchers doing animal feeding trials with mycotoxins. Diagnostic testing for mycotoxins in contaminated feedstuffs was improved utilizing affinity column methodology in combination with HPLC analysis. A three week (hatch to 21 days) study was conducted to evaluate the efficacy of turmeric (Curcuma longa) powder (TMP), a source of the antioxidant curcumin (1.48%), and a hydrated sodium calcium aluminosilicate (HSCAS) adsorbent to ameliorate aflatoxicosis in broilers. Compared with chicks fed AFB1, the addition of TMP or HSCAS to the AFB1 diet improved (P<0.05) growth performance of chicks. Turmeric and HSCAS also ameliorated (P<0.05) aflatoxicosis in terms of serum protein, albumin, cholesterol, and calcium. The decreased antioxidant function in terms of level of peroxides, superoxide dismutase activity and total antioxidants in liver due to AFB1 were also alleviated (P<0.05) by the inclusion of TMP and/or HSCAS. The reduction in the severity of hepatic microscopic lesions due to TMP and HSCAS inclusion in the AFB1 diet demonstrated the protective action of curcumin/HSCAS used in the present study. A three week study (hatch to 21 days) was conducted to evaluate the efficacy of total curcuminoids (TCMN) to ameliorate the effects of aflatoxin (AF) in broilers. Food grade turmeric powder (Curcuma longa) containing 2.55% TCMN was the source of TCMN. The addition of 74 and 222 ppm TCMN to the AF diet improved (P<0.05) weight gain and feed efficiency. The increased (P<0.05) relative liver weight in chicks fed AF was reduced significantly (P<0.05) by 74, 222, and 444 ppm TCMN. The inclusion of 222 ppm TCMN ameliorated the adverse effects of AF on serum total protein, and albumin and ³-glutamyl transferase activity. Decreased antioxidant functions in liver due to AF feeding were alleviated by the inclusion of 222 ppm TCMN. Addition of 222 ppm TCMN to the AF diet provided the greatest protection against AF and demonstrated maximum antioxidant activity. An experiment was conducted to determine the efficacy of the adsorbent Calibrin A (CA), in ameliorating the toxic effects of aflatoxin (AF) in broiler chicks. A second objective was to determine if Calibrin A at 0.5% of the diet would negatively affect chick performance. The addition of CA to chick diets at a level of 0.50% did not negatively affect (P > 0 .05) chick performance or relative liver weight or cause any liver lesions. Compared with controls, feed intake (FI) and body weight gain (BWG) were depressed (P < 0.05) in chicks fed AF, with greater reductions in FI and BWG observed in birds fed 3 mg/kg AF compared with those fed 2 mg/kg AF. The addition of 0.25% or 0.50% CA to the AF-contaminated diets significantly (P < 0.05) improved FI and BWG. Compared with controls, relative liver weights were higher in chicks fed AF (P < 0.05), and the addition of CA (0.25% or 0.5%) to the AF diet containing 2 mg/kg AF reduced (P < .05) the increase in liver weight. Compared to controls with a lesion score of 1 (no lesions), liver lesion scores in birds fed AF averaged 2.69 (moderate aflatoxicosis). The addition of 0.25 or 0.50% CA to the 2 mg/kg AF diet reduced the lesion scores to 2.25 and 1.63, respectively. Results indicate that CA at 0.50% of the diet did not negatively affect chick performance, relative liver weight, or cause any liver lesions indicating that this level of CA did not negatively affect nutrient content of the diet. CalibrinA at 0.25 or 0.5% of the diet significantly ameliorated the toxic effects of 2 and 3 mg/kg AF in young growing chicks. The North Dakota group reported on natural antifungal components of flax seed were evaluated as potential additives to prevent mold growth in products such as pasta. The Penn State group conducted growth chamber studies to examine the role of temperature in fungal colonization and deoxynivanol development when Fusarium graminearum, the causal agent of Head Scab of Wheat, is inoculated on wheat heads. When inoculated plants are kept at 15 C fungal growth is limited but toxin is produced at significant levels and is translocated in the wheat head to areas that are not yet colonized by the fungus. Objective 4. Define the regulation of mycotoxin biosynthesis and the molecular relationships between mycotoxigenic fungi The Michigan group reported that the trichothecene mycotoxin deoxynivalenol (DON; vomitoxin) is produced by several Fusarium species during infection of grain crops. DON acts as a virulence factor in wheat. Previous experiments, including Affymetrix GeneChip assays, indicated that genes involved in DON biosynthesis are highly expressed during infection of wheat and barley. Notably, DON biosynthesis gene expression appears to be correlated with the infection front, and to diminish once the fungus is fully established. In order to better understand the role of DON during plant infection, we chose to monitor expression of Tri5, the trichodiene synthase gene, necessary for DON production. We dissected wheat heads for several days following inoculation with Fusarium graminearum and monitored Tri5 expression using quantitative reverse-transcript PCR (qRT-PCR). The highest proportional TRI5 expression was consistently detected at the growing front, in accordance with the microarray data. However, during the course of this study, TRI5 gene expression was never lost, i.e. hyphae in kernals nearest the inoculation point did not cease expressing TRI5. We are continuing to study expression of TRI5 through kernel development in order to understand how and when DON accumulates. In addition, we are beginning to test the correlation between green tissue and TRI5 expression by treating the wheat with the strobilurin fungicides. Strobilurins delay plant senescence, and have been implicated in increased DON accumulations. By determining the precise interaction between strobilurins and DON, we may be able to better time fungicide application to reduce or eliminate this synergism. The Penn State group reported that molecular genetics studies of mycotoxigenic fungi often require the use of classical genetic analysis. We confirmed that F. verticillioides isolate M-3125 (mat -) carries the spore killer sensitive allele. The genome sequence for this isolate is publically available and as such is frequently used for molecular genetic analysis. The presence of the spore killer sensitive allele in M-3125 means that in crosses with isolates carrying the killer allele (the majority of isolates in nature and in culture collections) genes with linkage to spore killer will not be recovered in the cross confounding genetic analysis. We searched a collection of fifty F. verticillioides isolates from the Penn State Fusarium Research Center for mat + isolates that were either spore killer sensitive. Two isolates, M-7815 and M-8024, meet these criteria and can be used in crosses with M-3125 without the segregation distortion that accompanies crosses with spore killer isolates The Purdue group reported that Expression of AREA also appears to affect the ability of F. verticillioides to colonize maize kernels. Indiana group used strains containing the reporter gene of ²-glucuronidase (GUS) fused to the promoter of NIAD (nitrate reductase) to study AREA expression and fungal growth. Because NiaD is transcriptionally regulated by AreA, it was possible to measure AREA expression by the activation of the NIAD promoter in the reporter construct. Growth, GUS expression, and fumonisin were monitored in a wild type, a disruption mutant (areA), and a strain with constitutive expression of AREA. While F. verticillioides grows equally well on each of the developmental stages of maize kernels (blister, milk, dough, and dent), AREA expression was found to be the lowest in colonized blister stage kernels and to increase with kernel maturity. Strain areA grew poorly on mature maize kernels, but grew similar to the wild type when provided with the addition of ammonium phosphate. FB1 was not produced by strain areA under any condition or by the wild type with added ammonium phosphate. However, constitutive expression of AREA rescued the growth and FB1 defects in areA. The strain with constitutive expression of AREA also produced FB1 on maize kernel with added ammonium phosphate. Growth of areA on the blister stage kernels was similar to the wild type, indicating that utilization of the available nitrogen source in these kernels does not require AREA. GUS activity was not detected in the wild type, suggesting that nitrogen metabolite repression of NIAD occurs during growth on blister kernels. GUS expression was detectable in constitutive-expression strain, which supports the conclusion that blister stage kernels are nitrogen metabolite repressive. These results provided some insight as to the effects that carbon metabolism and carbon:nitrogen ratio have on secondary metabolism during colonization

Publications: Akare, S. Hsiao, S-H, Volmer, P.A., Wallig, M. A., and Haschek, W.M. 2008. Fumonisin induced pulmonary edema in pigs: a case report. Tox Path 36:163-164, P43. Bluhm, B. H., Kim, H., Butchko, R. A. E., and Woloshuk, C. P. 2008. Involvement of ZFR1 of Fusarium verticillioides in kernel colonization and the regulation of FST1, a putative sugar transporter gene required for fumonisin biosynthesis on maize kernels. Molecular Plant Pathology 9:203-211. Carter, L. L. A., J. F. Leslie & R. K. Webster. 2008. Population structure of Fusarium fujikuroi from rice and water grass in California. Phytopathology 98: 992-998. Coulibaly, O., K. Hell, R. Bandyopadhyay, S. Hounkponou & J. F. Leslie. 2008. Economic impact of aflatoxin contamination in Sub-Saharan Africa. In: Mycotoxins: Detection Methods, Management, Public Health and Agricultural Trade (J. F. Leslie, R. Bandyopadhyay & A. Vis-conti, eds.), pp. 67-76. CABI, Kew, UK. 476 pp. Dakovic, A., Matijasevic, S., Rottinghaus, G. E., Ledoux, D. R., Butkeraitis, P., and Sekulic, Z. 2008. Aflatoxin B1 adsorption by natural and copper modified montmorillonite. Colloids and Surfaces B: Biointerfaces 66:20-25. Delgado, J.A., Schwarz, P.B., Gillespie, J., Rivera-Varas, V., Secor, G.A. 2008. Presence and distribution of deoxynivalenol in potato tubers inoculated with Fusarium graminearum. Phytopathology. 98:S45. Gowda, N. K. S. and D. R. Ledoux. 2008. Use of antioxidants in amelioration of mycotoxin toxicity: A Review. Animal Nutrition and Feed Technology 8:1-11. Gowda, N. K. S., Ledoux, D. R., Rottinghaus, G. E., Bermudez, A. J., and Chen, Y. C. 2008. Efficacy of turmeric (Curcuma longa) and a hydrated sodium calcium aluminosilicate to ameliorate the adverse effects of aflatoxin in broiler chicks. Poultry Science 87:1125-1130. Gupta, S., Jindal, N., Khokhar, R. S., Asrani, R. K., Ledoux, D. R., and Rottinghaus, G. E. 2008. Individual and combined effects of ochratoxin A and Salmonella enteric serovar Gallinarum infection on pathological changes in broiler chickens. Poultry Science 37(3):265-272. Hallen, H., and F. Trail. 2008. The L-type calcium ion channel, Cch1, affects ascospore discharge and mycelial growth in the filamentous fungus Gibberella zeae (anamorph Fusarium graminearum). Eukaryotic Cell. 7:415-424. Haschek, W.M. and Tumbleson, M.E.. Fumonisin toxicosis in domestic animals: Current knowledge and research needs. 2008. Midwest Section of AOAC International Annual Meeting and Exposition, Bozeman, MT, June 10, 2008. Haschek W.M., Hsiao S-H, Wicklow, W.T. 2008. The fungal metabolite, pyrrocidine A, induces apoptosis in HepG2 hepatocytes and PK15 renal cells. The Toxicologist CD  a supplement to Toxicological Sciences 86 (S-1): Abs. 2227. Hill, N.S., Neate, S.M., Cooper, B., Horsley, R., Schwarz, P., Dahleen, L.S., Smith, K.P., O'Donnell, K. and Reeves. J. 2008. Comparison of ELISA for Fusarium, visual screening, and deoxynivalenol analysis of Fusarium head blight for barley field nurseries. Crop Sci. 48:1389-1398. Hye-Young Yu, Jeong-Ah Seo, Kap-Hoon Han, Won-Bo Shim, Sung-Hwan Yun, and Yin-Won Lee. 2008. Functional analyses of heterotrimeric G protein Ga and Gb subunits in Gibberella zeae. Microbiology 154: 392-401. Kim, H., and Woloshuk, C. P. 2008. Role of AREA, a regulator of nitrogen metabolism, during colonization of maize kernels and fumonisin biosynthesis in Fusarium verticillioides. Fungal Genet. Biol. 45:957-953. Kottapalli, B. and Wolf-Hall, C. 2008. Effect of hot water treatments on the safety and quality of Fusarium-infected malting barley. International Journal of Food Microbiology. 124:171-17. Kuldau, G. and Bacon, C. 2008. Clavicipitaceous endophytes: Their ability to enhance grass resistance to multiple stresses. Biological Control 46: 57-71. Lee, J., J. E. Jurgenson, J. F. Leslie & R. L. Bowden. 2008. Alignment of genetic and physical maps of Gibberella zeae. Applied and Environmental Microbiology 74: 2349-2359. Lee, J., J. F. Leslie & R. L. Bowden. 2008. Expression and function of sex pheromones and receptors in the homothallic ascomycete Gibberella zeae. Eukaryotic Cell 7: 1211-1221. Lee, J., J. F. Leslie & R. L. Bowden. 2008. Functions of the sex pheromones in Gibberella zeae. Rivista di Patologia Vegetale 90: S3.26. Leslie, J. F., R. Bandyopadhyay & A. Visconti, eds. 2008. Mycotoxins: Detection Methods, Management, Public Health and Agricultural Trade. CABI, Kew, UK. 476 pp. Leslie, J. F., and R. L. Bowden. 2008. Fusarium graminearum: When species concepts collide. Cereal Research Communications 36 (suppl. B): 609-615. Leslie, J. F., J. Lee, J. E. Jurgenson & R. L. Bowden. 2008. An update of the genetic map of Gibberella zeae. Rivista di Patologia Vegetale 90: S3.26. Lima, C. S., S. S. Costa, M. A. Campos, J. F. Leslie & L. H. Pfenning. 2008. Etiology of mango malformation and PCR detection of its causal agent in Brazil. Rivista di Patologia Vegetale 90: S3.65. Mansfield, M. A., Jones, A. D. and Kuldau, G. A. 2008. Contamination of fresh and ensiled maize with four mycotoxins produced by Penicillium species. Phytopathology 98: 330-336. Manthey, F., Sinha, S, Hall III, C., Wolf-Hall, C.E. 2008. Effect of flaxseed on flour and packaging on shelf-life of refrigerated pasta. Journal of Food Processing and Preservation. 32:75-87. Minnaar-Ontong, A., L. Herselman, W. M. Kriel & J. F. Leslie. 2008. Population dynamics of Fusarium head blight in South Africa. Rivista di Patologia Vegetale 90: S3.66. Sasanya, J., Hall III, C., and Wolf-Hall, C. 2008. Quantification of deoxynivalenol, masked deoxynivalenol and Fusarium graminearum pigment in hard red spring wheat samples using a new LC-UV/MS method. J. Food Prot. 71:1205-1213. Settivari, R. S., Evans, T. J., Rucker, E., Rottinghaus, G. E., and Spiers, D. E. 2008. Effect of ergot alkaloids associated with fescue toxicosis on hepatic cytochrome P450 and antioxidant proteins. Toxicology and Applied Pharmacology 227 (3):347-356. Settivari, R. S., Evans, T. J., Eichen, P. A., Rottinghaus, G. E., and Spiers, D. E. 2008. Short-and long-term responses to fescue toxicosis at different ambient temperatures. J Thermal Biology 33:213-222. Sharma. D., Asrani, R. K., Ledoux, D. R., Jindal, N., Rottinghaus, G. E., Bermudez, A. J., and Gupta, V.K. 2008. Individual and combined effects of fumonisin B1 and moniliformin toxicity on clinicopathological and cell mediated immune response in Japanese quail. Poultry Sci 87:1039-1051. Shelton, B. G. & J. F. Leslie. 2008. Comparative risks of airborne and foodborne molds and mycotoxins. In: Mycotoxins: Detection Methods, Management, Public Health and Agricultural Trade (J. F. Leslie, R. Bandyopadhyay & A. Visconti, eds.), pp. 317-324. CABI, Kew, UK. 476 pp. Stewart, Jr. R. L., Scaglia, G., Abaye, O. A., Swecker, Jr. W. S., Rottinghaus, G. E., Boland, H. T., McCann, M., and Fontenot, J. P. 2008. Estimation of forage intake by steers grazing three fescue types and determination of alkaloids in ruminal fluid and forage. Professional Animal Scientist 24:578-587. Suanthie, Y., Cousin, M.A. and Woloshuk, C. P. Multiplex real-time PCR for detection and quantification of mycotoxigenic Aspergillus, Penicillium, and Fusarium. J. Stored Prod. Res. doi:10.1016/j.jspr.2008.12.001 Wu. X. 2008. Deoxynivalenol alters circulating and splenic leukocyte and cell migration markers in interaction with time course and sex in young and old BALB/c mice. PhD Dissertation, Iowa State University Library, Ames, IA. Xu, Y, Hall III, C., Wolf-Hall, C. 2008. Fungistatic activity of heat-treated flaxseed determined by response surface methodology. J. Food Sci. 73:M250-M256. Xu, Y., Hall III, C., Wolf-Hall, C, Manthey, F. 2008. Fungistatic activity of flaxseed in potato dextrose agar and a fresh noodle system. International Journal of Food Microbiology. 121:262-267. Xu, Y, Hall III, C., Wolf-Hall, C. 2008. Antifungal activity stability of flaxseed protein extract using response surface methodology. J. Food Sci. 73:M9-M14. Yigezu, Y., Alexander, C., Preckel, P.V., Maier, D.E., Woloshuk, C.P., Mason, L.J., Lawrence, J. and Moog, D. 2008. Optimal Management of Molds in Stored Corn. Agricultural Systems 98:220-227. Yoon-E Choi, Jillian Brown, Courtney Williams, Lorena Canales, and Won-Bo Shim. 2008. GAC1, a gene encoding a putative GTPase-activating protein, regulates bikaverin biosynthesis in Fusarium verticillioides. Mycologia 100: 695-703. Yoon-E Choi and Won-Bo Shim. 2008. Identification of genes associated with fumonisin biosynthesis in Fusarium verticillioides via proteomics and quantitative real-time PCR. Journal of Microbiology and Biotechnology 18: 648-657. Yoon-E Choi and Won-Bo Shim. 2008. Enhanced homologous recombination efficiency in Fusarium verticillioides by disruption of FvKU70, a gene required for a non-homologous end joining mechanism. The Plant Pathology Journal 24: 1-7. Yoon-E Choi and Won-Bo Shim. 2008. Functional characterization of Fusarium verticillioides CPP1, a gene encoding a putative protein phosphatase 2A catalytic subunit. Microbiology 154: 326-336. Zhang, H., Wolf-Hall, C. and Hall III, C. 2008. Modified microwave-assisted extraction of ergosterol for measuring fungal biomass in grain cultures. J. Agri. Food Chem. 56: 1107711080. Zhou, B., He, G.Q., Schwarz, P.B. 2008. Occurrence of bound deoxynivalenol in Fusarium head blight-infected barley (Hordeum vulgare L.) and malt as determined by solvolysis with trifluoroacetic acid. J. Food Prot. 71: 1266-1269.