Performing Department
Veterinary Medicine
Non Technical Summary
Develop and validate new advanced multi-matrix detection methods that can be used by public agencies, and by private diagnostic labs to provide mycotoxins analysis services to food industries. These technologies will replace some of the current methodologies with high sensitivity, accuracy and efficiency.Help to generate new protocols for monitoring mycotoxins in agricultural products that can be adopted by biosecurity agencies concerned with potential bioterror attacks on the nation.
Animal Health Component
0%
Research Effort Categories
Basic
80%
Applied
20%
Developmental
(N/A)
Goals / Objectives
Objective 2: Establish integrated strategies to manage and reduce mycotoxin contamination in cereals and in forages.
Project Methods
Method 1: SurveillanceSurveillance 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 detectFusarium,Penicillium, andAspergillus. 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.Method 2: InterventionGrains 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.Method 3: Fungal Population GeneticsMolecular 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 ofFusarium graminearum). Members from KS and KY will lead an effort to study segregation of quantitative traits involved in fecundity, aggressiveness, and mycotoxin production inF. 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.