US 12,274,268 B2
Methods and compositions for increasing infectivity of entomopathogenic nematodes
David I. Shapiro Ilan, Macon, GA (US); and Fatma Kaplan, Davis, CA (US)
Assigned to The United States of America, as represented by the Secretary of Agriculture, Washington, DC (US); and Pheronym Inc., Davis, CA (US)
Filed by The United States of America, as represented by the Secretary of Agriculture, Washington, DC (US); and Pheronym Inc, Davis, CA (US)
Filed on Feb. 25, 2020, as Appl. No. 16/800,745.
Claims priority of provisional application 62/814,364, filed on Mar. 6, 2019.
Prior Publication US 2020/0281213 A1, Sep. 10, 2020
Int. Cl. A01N 63/12 (2020.01)
CPC A01N 63/12 (2020.01) 6 Claims
 
1. A method of producing a stable dry nematode infectivity-increasing composition which comprises:
(i) obtaining a nutrient-depleted nematode growth medium by growing non-pathogenic bacterivore nematodes, insect nematodes, or entomopathogenic nematodes in a growth medium selected from liquid broth, agar medium, other solid substrate, or at least one insect host cadaver;
(ii) producing an alcohol-nutrient-depleted nematode growth medium mixture by adding alcohol to said nutrient-depleted nematode growth medium to a final concentration of between about 10% to about 90% of said alcohol in said nutrient-depleted nematode growth medium and shaking for about 10 minutes at room temperature;
(iii) centrifuging said alcohol-nutrient-depleted nematode growth medium mixture to remove solid or insoluble matter while maintaining a supernatant from said centrifuging;
(iv) drying the supernatant from said centrifuging using a rotary evaporator to produce a dry extract;
(v) resuspending said dry extract in an aqueous medium to produce a water-soluble pheromone extract;
(vi) centrifuging said water-soluble pheromone extract to remove water-insoluble compounds while maintaining a water-soluble supernatant; and
(vii) drying said water soluble supernatant;
to produce a stable dry nematode infectivity-increasing composition.