MYCOPESTICIDES Collecting and Cultivating Entomopathogens Tradd Cotter Mushroom Mountain, LLC Liberty, SC

Myco- “mushroom, fungus” Pesticide- “substance that prevents, destroys, or mitigates any pest”

Types of Pests Insects – structural, human disease vectors, biting, agricultural Microbial – bacterial and fungal pathogens related to human and plant health Rodents

Types of Pests Insects – structural, human disease vectors, biting, agricultural Microbial – bacterial and fungal pathogens related to human and plant health Rodents - Monsanto, DOW, Syngenta, etc

Concerns with Mycopesticides Some insects are beneficial at one stage and a pest at another. Caterpillars eating agricultural crops Adult butterflies serve as beneficial pollinators

Concerns with Mycopesticides Host jumping Range of specificity must be determined within a clade of family of insects Mechanisms for target specificity, spore adhesion and host recognition not yet understood Residual effects

Benefits of Mycopesticides Narrow bandwidth of target specificity using cloned ecotypes. Self replicating – inexpensive to manufacture Low to no ecotoxicity compared to chemical pesticides Can be collected from native or local stock to target local pests

Copyright – Tradd Cotter

7.5 BILLION POUNDS OF PRODUCT USE ACTIVE INGREDIENT CHLORPYRIFOS 7.5 BILLION POUNDS OF PRODUCT USE PER YEAR

Metarhizium feeding on Varroa Mites http://www.ars.usda.gov/is/graphics/photos/oct04/k11145-17i.jpg

Isolation and Purification of Cultures Insect Surface Sterilize insect – ethanol / sterile water wash – alternate 3 times 30 seconds each Open insect and remove tissue in appendages, not abdomen or gut Use dental tool or needle – peroxide wash tissue for 5 minutes if necessary Use antibiotic media

Isolation and Purification of Cultures Teleomorph – Fruiting Body Place insect and attached fungus into a humidity chamber to allow maturation of spore bearing surface Use needle to touch emerging spores from parethecia (pits) OR surface sterilize fruiting body and remove sterile tissue

Isolation and Purification of Cultures Maintaining Entomopathogens Capture target insects specific for fungus Blend exoskeletons into agar formulation before sterilization Host recognition and target specificity begins to deteriorate if host is not present after several transfers – lose interest Submerged agar storage on slants Liquid combined with glycerol and frozen

Death and Mummification MYCOPESTICIDES WHAT WE HAVE ACCOMPLISHED Discovery Isolation of Pure Culture Ant Gallery Construction Death and Mummification ( 7 days ) Addition of Fungus ( 5 days )

MYCOPESTICIDES TARGET-SPECIFIC, Biological alternative to chemicals Broad Range Acephate Toxicity Chemical Pesticide Large Target Window MYCOPESTICIDAL Narrow Target Window

MYCOPESTICIDES PLAN OF ACTION AND CHALLENGES DNA AND PATENT PROTECTION Identify and File Delivery System or Trade Secret Process 2. PERMITTING AND FIELD TRIALS EPA & Clemson Pesticide Permitting Applied For INDUSTRIAL DEVELOPMENT Process development and scaling from parallel models LICENSE AND DISTRIBUTE Manufacture or license active ingredient