Sales Manager, North America Mycotoxins – Impact, Risk and Opportunity Chris Seward Sales Manager, North America

Mycotoxins Toxin Origin Deoxynivalenol (DON) Fusarium sp. Fumonisin Aflatoxin Aspergillus sp. Toxin Origin Zearalenone Fusarium sp. T2-HT2 Ochratoxin Aspergillus sp. Penicillium sp.  Natural chemicals Produced by mold Can contaminate many kinds of crops Stable, don’t break down over time and can withstand high temperature / heat

Conditions That Contribute To Mycotoxin Contamination Weather Conditions Environmental Handling / Equipment Weather: Drought, Too Much Rain, Excessive Heat or Cold can stress the crop Environmental & stress conditions: Weed Competition, Insect damage Poor Fertilization, High Crop Density can weaken the plant’s natural defenses and promote colonization by mycotoxin-producing fungi as well as toxin-formation. Handling / Equipment: Make sure it is in good operation condition and cleaned. Damage can lead to mycotoxin issues. Storage: Harvesting timing – Moisture content and water activity can have a large impact on mold growth and thus mycotoxin production. Make sure storage facility is appropriately ventilated and controlled for pests. Storage

Mycotoxins Are Found Around The Globe https://www.researchgate.net/figure/Distribution-of-mycotoxins-world-map-Mycotoxin-Survey-Program-2011_fig1_305943346

Co-occurrence of mycotoxins is common https://www.google.com/search?q=sources+of+mycotoxin+contamination+infographic&biw=1536&bih=711&tbm=isch&source=iu&ictx=1&fir=SsfpBfmvoH37EM%253A%252Cls9bTOOqBpkpsM%252C_&usg=__8wdCVqRYzFTtlXHGolUT_eoxSD0%3D&sa=X&ved=0ahUKEwiti4evn8TZAhWCw4MKHdYlCNcQ9QEIVTAF#imgrc=LIdHXZRScEaOzM: This is data from the Americas (but the story is the same globally)

Economic Impact Of Mycotoxins Economic Loss Occurs Due To Yield loss due to disease induced by toxigenic fungi Reduce crop value resulting from mycotoxin contamination Loss in animal productivity due to mycotoxin –related health problems Human health costs Additional Talking points: 1. Depending on the study, estimates of the economic cost of mycotoxin contamination are upward of $1 billion in the US alone. 2. It is believed that in developing countries the losses are greater than those calculated in the United States. Mycotoxins in Crops: A Threat to Human and Domestic Animal Health David G. Schmale III Virginia Polytechnic Institute and State University, Blacksburg, VA Gary P. Munkvold Iowa State University, Ames, IA Source: Mycotoxin in Crops: A Threat to Human and Domestic Animal Health David G. Schmale III and Gary P. Munkvold url: https://www.apsnet.org/edcenter/intropp/topics/Mycotoxins/Pages/EconomicImpact.aspx

Economic Impact Of Mycotoxins Aflatoxins Crops Impacted Primary Animals Impacted Effects Liver Damage Intestinal Bleeding Cancer Corn Peanuts Cottonseed Tree nuts Dairy Products

Economic Impact Of Mycotoxins Vomitoxin / Deoxynivalenol (DON) Crops Impacted Primary Animals Impacted Effects Feed refusal Diarrhea Vomiting Skin disorders Reduced Growth Wheat Barley Oats Corn

Economic Impact Of Mycotoxins Fumonisins Crops Impacted Primary Animals Impacted Effects Pulmonary edema Esophageal cancer Neural tube defects Liver damage Reduced growth Corn Silage

How Do Mycotoxins Impact Business Impact of Mycotoxin Contamination Lower egg production & quality Reduced Fertility Decreased feed efficiency and weight gain Increased mortality rates Suppression of immunity Poor conception Reduced meat / milk production Suppression of immunity Organ damage Reduced Fertility / Abortion Poor growth rates https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4988553/ Mycotoxins can impact individually or additively http://www.mycotoxins.info/en/effects/pigs/ Impact on pigs http://www.knowmycotoxins.com/sites/default/files/fact-sheets/mycotoxin-issues-in-beef-flyer.pdf Infertility Organ damage Cancer

Testing Produces Data That Drives Business Decisions Without Testing, No Information To Support Segregation / Acceptance Testing Provides Information To Drive Business Decisions Also provided data to support any legal action should it arise $$$$ $$$ $$ $ -$$ Not hitting contract specs; incur additional transportation expense Taking lesser contracts to avoid rejection; not getting maximum value for ‘good’ grain +$$ Ability to hit a wide variety of contract specs; opens more markets; reduces transportation costs

How Do Mycotoxins Impact Business (Corn) Discuss that every point in this map there is value added. It is important to have myco information before adding value to any product. Why would you add value to a product that you know isn’t going to meet the market need? Test before. Also just because you don’t test doesn’t mean your customer wont – don’t get caught by surprise after you have already done all the work. Fast accurate answers are key to driving operational efficiencies and a healthy bottom line Fast, Accurate, Decision Point Answers Are Key To Driving Operational Efficiencies And A Healthy Bottom Line

Mycotoxin Testing Methods Rapid Methods: Fluorometry ELISA LFDs Research: Academic, government, commercial labs Finished food, biologicals HPLC LC-MS/MS Quantitative Lab-Based: Commercial/official control labs Finished food, feed, complex matrices, multi-toxin analyses Field testing: High volume, short turnarounds at receiving and transit points, processors Commodities and feed components

No official definition of “rapid” – but for our purposes: Rapid Testing Methods No official definition of “rapid” – but for our purposes: Simple and easy to use Relatively fast (minutes vs hours) Capable of testing mycotoxins outside of a laboratory setting For this review: testing methods capable of detecting mycotoxins in a single, pre-ground sample within 30 minutes “Assay time” starts with this pre-ground sample Extraction steps Pipetting/transferring extract and various buffers Development time Read time

Fluorometric Assay Requires sample ‘clean-up’ to remove interferences Can be fully quantitative Requires fluorometer for interpretation Assay time 25-35 minutes

ELISA Enzyme linked immuno-sorbent assay – microtiter plate – competitive ELISA – antibody/antigen complex Measured with ELISA reader Requires fairly sophisticated operators, and grouping tests rather than allowing serial testing Can be fully quantitative with the use of calibrators Highly specific, highly sensitive, but can be implemented for use with complex matrices Limited efficiency for inbound grain Assay time 15-25 minutes+

Lateral Flow Devices (LFDs) Uses the same ELISA principles (antibody/antigen complex, competitive assay) on a backed membrane Visual tests with specific cutoffs give qualitative results More recently, tests utilizing readers can interpret and return quantitative results Low technical comprehension and ability needed User-friendly, steady throughput (no “queue and batch” of samples for testing) Assay time 3 - 10 minutes

Errors Can Be Introduced At Various Steps In The Testing Process Run, Read & Record Sample Extract Dilute Reduce errors and by default you improve accuracy (simple protocols are better)

Errors Can Be Introduced At Various Steps In The Testing Process Run, Read & Record Sample Extract Dilute Sampling of the Truck Probe needs to be fully functional and cleaned. Sampling protocol needs to be adequate enough to get a representative sample of the container both (There are some good sampling protocols out there – this is one from GIPSA)

Errors Can Be Introduced At Various Steps In The Testing Process Run, Read & Record Sample Extract Dilute Subsampling: Key to making sure that you get a representative sample is using a diverter to randomly divide the sample into equal parts (and ensure mixing) Depth, number of spots sampled make sure you purge the probe between trucks.

Errors Can Be Introduced At Various Steps In The Testing Process Run, Read & Record Sample Extract Dilute Grinding: Grind enough sample incase a retest is needed. Grind to the correct size to maximize extraction efficiency Make sure the grinder is clean to avoid cross contamination

Errors Can Be Introduced At Various Steps In The Testing Process Run, Read & Record Sample Extract Dilute Weighing out the appropriate amount Make sure your scale is calibrated and up to spec Add correct amount of water – keep ratios as defined in published protocols. Shake for exact amount of time required

Errors Can Be Introduced At Various Steps In The Testing Process Run, Read & Record Sample Extract Dilute Pipettes must be clean and calibrated Careful pipetting

Errors Can Be Introduced At Various Steps In The Testing Process Run, Read & Record Sample Extract Dilute Timing of the assay is important don’t over or under run the assay Visual examination of the assay can lead to miss reading depending the eye of the examiner. Levering an instrument based system can help eliminate some subjectivity. Hand recorded results can sometime be difficult to read, or find and can sometimes be incomplete. Leverage a software system that can help populate fields will lead to more consistent language, easier access to historical results and more compete records.

Common Extraction & Common Run Time Can Help Reduce Some Of These Errors…

…By Consolidating Steps Across Multiple Assays Run, Read & Record Sample Extract Dilute That Allows You To Test DON, Afla & FUM Use One Sample To Create One Extract And Run, Read & Record at One Time Use this slide to tee up the next slide about speed Simplicity reduces error and helps speed up common extraction. This is less than 3 minutes per assay. And in many cases speed is key to improving your operational efficiency 3 Results in 8 Minutes

Speed Key To Improving Operational Efficiency

THANK YOU FOR THIS OPPORTUNITY AND THANK YOU FOR YOUR TIME AND ATTENTION