Multidisciplinary Approaches to Reduce the Impact of Corn Aflatoxin on Texas

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1 Multidisciplinary Approaches to Reduce the Impact of Corn Aflatoxin on Texas Dr. Joseph Awika Food Technology Dr. David Baltensperger Soil and Crop Sciences Dr. Youjun Deng Soil Mineralology Dr. Joe Dixon Soil Mineralology Dr. Tim Herrman Grain Science / State Chemist Dr. Tom Isakeit Plant Pathologist Dr. Mike Kolomiets Molecular Plant Pathology Dr. Kyung-Min Lee Cereal Chemistry Mr. Kerry Mayfield Corn Breeding Dr. Seth C. Murray Corn Breeding Dr. Gary Odvody Plant Pathologist (Corpus Christi) Dr. Tim Phillips - Toxicology and Food Science Dr. Wenwei Xu Corn Breeding (Lubbock) Funding: Texas AgriLife, Texas Corn Producers Board, USDA, USAID, NSF, many others.

2 Dr. Gary Odvody What is Aflatoxin? - Highly toxic compound produced by fungus Aspergillus flavus - Toxicity at low concentrations: - 20ppb action level / above 500ppb must be destroyed - Potent carcinogen - Pre-harvest production in corn kernels - Can also occur on corn in storage

3 Interaction Between Pathogen, Environment and Plant Infection / resistance Vs. aflatoxin production Aspergillus flavus spores Suitable environment Kerry Mayfield Disease Triangle Susceptible host: corn & others

4 Environments Favorable for Aflatoxin - Drought / heat stress - High temperature/ low soil moisture - Especially after silking enter silks - Corn maturation into increasing heat/ stress - Insects - Primarily Lepidotera - Stalk borers - Corn earworm (CEW), Fall armyworm (FAW) - Wounding - Aflatoxin accumulation variable: - Within an ear - maybe a few kernels - Across a field drought hot spots, field edge - Late planting increases stress Dr. Gary Odvody

5 Corn Yields and Aflatoxin Aflatoxin Loss Liability 2008 Office of the State Chemist Sampling 2008 Kerry Mayfield

6 Aflatoxin levels in corn 2009 Office of the Texas State Chemist 20 ppb ppb 300 ppb 500 ppb Dr. Tim Herrman & Dr. Kyung-Min Lee * The color indicates the highest reported incidence in a county.

7 Integrated Pest Management identify agronomic practices that reduce A. flavus. Plant Breeding find and incorporate genes or traits for A. flavus resistance into hybrids. Plant Pathology identify disease factors, develop competing atoxigenic strains. Remediation Find binders or processing methods to reduce the toxin or toxin absorbance. Food Science/ Veterinary Nutrition Identify processing methods that can reduce aflatoxin.

8 Pre-Harvest Aflatoxin Contamination Texas 2008 corn farm gate value = $1billion (13 th in Country) $14 million loss liability from mycotoxins in 2008 Most elite US germplasm is highly susceptible in Texas Breeding is one of the least expensive and least intensive methods for reducing aflatoxin contamination No identified sources of complete resistance to Aspergillus / aflatoxin Test in multiple environments with inoculation Percent (%) Acreage with Aflatoxin Claims 2008

9 Aflatoxin Resistance Differs Across Corn Inbred Lines and Hybrids Weslaco Aflatoxin (ng g -1 ) Genotype Pedigree 6 (Tx772/CML326)-B-B5-B-B/ Tester 8 LAMA B-B/ Tester Mayfield et al, 2007 Genotype Kerry Mayfield

10 Corn Breeding Pyramid for Reduced Pre-Harvest Aflatoxin Contamination How do we separate these various correlated traits? Stress Resistance Row number - Husk Coverage - Long, cover tips - Tight and thick - Maturity - Flowering time - Days to maturity - Kernal hardness - Ear nod (ear droop) - Earworm resistance - Drought tolerance Ear length Stay Green YIELD Lodging Kernel size Kernel density

11 QTL Mapping - RIL Hybrids 135 RIL Testcrosses (B73o2 x CML 161) College Station, Weslaco and Corpus Christi 3 Replications 15 QTL detected in traits Kerry Mayfield WE Oil CS Oil CC Oil CS Protein WE Nodded Ears CS Starch CS Log AF WE Log AF

12 Aflatoxin levels can be reduced by deleting a lipoxygenase gene LOX5 in corn under field conditions in different genetic backgrounds natural diversity / non-gmo Tx714 FR2128 Tx772 Dr. Mike Kolomiets

13 Corn ears: rep 5 showing relative yield and insect damage at Corpus Christi, TX in Conventional hybrids are shown by row and traits are shown in columns Avg. 646 ppb Avg. 898 ppb Avg ppb VT Pro Yieldgard CB Non-Bt Hyb 1 Hyb 2 Hyb 3 Dr. Gary Odvody

14 Dr. Tom Isakeit

15 Bentonites as Adsorbents Economically feasible Socially acceptable Environmentally friendly Ines Mulder Maria Guadalupe Tenorio Arvide Dr. Youjun Deng & Dr. Joe Dixon

16 Testing Natural Bentonites for Adsorption Sites and Ability 41 samples USA (TX, WY, MS, ID), Australia, Brazil, Japan, Korea, Mexico, Mozambique, Spain, Turkey, South Africa Poultry test (on going) D-spacing & Adsorption Sites Dr. Youjun Deng & Dr. Joe Dixon

17 Dioctahedryl Smectite (clay-based enterosorption) therapies for aflatoxins and other food contaminants. - No adverse health effects from consumption of NovaSil clay Animal Feeding Protected animals from diet levels as high as 7,500 ppb. Human Supplement Reduced urine levels of AFM1 after 3-months intervention Deliverable Product Ghana Dr. Timothy Phillips

18 Sampling Procedures are Extremely Important and Limiting - Very Expensive to measure ( $8 to $100+ / sample) - Heterogenous within an ear, field, grain bin, shipping container To make progress: - Need increased testing speed, accuracy, and sample size while reducing costs - In research, best inoculation procedures must be followed

19 HPLC-Tandem MS Method (Aflatoxin) - Fast, compared with the tradition HPLC-Fluorescence method - Specific, quantitative, sensitive (LOQ 0.5 ppb) Diluted in 20% MeOH AF-STD MRM of 10 Channels ES+ 331 > 245 (AFG2) 3.00e4 % 0 AF-STD MRM of 10 Channels ES > 243 (AFG1) 2.29e MRM of 10 Channels ES > 259 (AFB2) 5.84e MRM of 10 Channels ES > 241 (AFB1) 5.06e5 % % % 0 AF-STD-7 0 AF-STD-7 0 AF-STD % MRM of 10 Channels ES TIC 8.23e Multiple reaction monitoring (MRM) of Aflatoxin B1, B2, G1 and G2 in LC/MS/MS Dr. Tim Herrman & Dr. Kyung-Min Lee Time

20 Increase Throughput of Testing With NIRS Aflatest is expensive and time consuming Need to screen 5,000-10,000 samples/ year Near Infrared Reflectance Spectroscopy (NIRS) Use NIRS calibration equation to predict contamination levels Develop calibration equation Identify informative samples from spectra Test 10% of samples for aflatoxin and/or Aspergillus Run samples on NIRS

21 NIRS Results Rsq. = Equation built using non-ag with a red (3/4) and val (1/4) sets 2,5,5,1 SCVD both vis and NIR (Rsq = ) Plot predicted vs. Full non-ag

22 Safety and Monitoring of Respiratory Aflatoxin and Particulates Apex Lite Personal Air Pumps were used to sample ~700L of air. Two of eight samples had detectible levels of respirable particulates (.1mg/m 3 ;.28mg/m 3 ) Clearvue Cyclone Air Cleaning

23 Aflatoxin Center of Excellence of the South Creating Linkages between: -States (Alabama, Georgia, Mississippi, North Carolina, Louisiana, Texas) - Commodity Groups - Public Research Institutions - Research, Education, Extension

24 Integrated Pest Management identify agronomic practices that reduce A. flavus. Plant Breeding find and incorporate genes or traits for A. flavus resistance into hybrids. Plant Pathology identify disease factors, develop competing atoxigenic strains. Remediation Find binders or processing methods to reduce the toxin or toxin absorbance. Food Science/ Veterinary Nutrition Identify processing methods that can reduce aflatoxin.

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