MYCOPESTICIDESCollecting and Cultivating Entomopathogens

Tradd CotterMushroom 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

CHLORPYRIFOS

7.5 BILLION POUNDS OF PRODUCT USE

PER YEAR

ACTIVE

INGREDIENT

http://www.ars.usda.gov/is/graphics/photos/oct04/k11145-17i.jpg

Metarhizium feeding on Varroa Mites

Isolation and Purification of CulturesInsect

• 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

MYCOPESTICIDESWHAT WE HAVE ACCOMPLISHED

Isolation of Pure Culture

Death and Mummification

( 7 days )

Discovery Ant Gallery Construction

Addition of Fungus ( 5 days )

MYCOPESTICIDESTARGET-SPECIFIC, Biological

alternative to chemicals

Chemical

Pesticide

Large Target Window

MYCOPESTICIDAL Narrow Target

Window

Broad Range Acephate Toxicity

MYCOPESTICIDESPLAN OF ACTION AND CHALLENGES

2. PERMITTING AND FIELD TRIALS

EPA & Clemson Pesticide Permitting Applied For

3. INDUSTRIAL DEVELOPMENT

Process development and scaling from parallel models

4. LICENSE AND DISTRIBUTE

Manufacture or license active ingredient

1. DNA AND PATENT PROTECTION

Identify and File Delivery System or Trade Secret Process