Water dispersible granule for agricultural applications

Abstract

The present disclosure provides a water dispersible granule (WDG) composition comprising yeast particles as a primary granule component and an organic liquid. The present disclosure further provides methods and processes for making and using the same. The WDG composition of this disclosure comprises yeast particles and an organic liquid and optionally comprises a pesticidal active ingredient. In embodiments, the WDG composition further comprises a dispersing agent and / or a wetting agent, and optionally a filler, and in embodiments does not contain a substantial amount of water. The water dispersible granule composition of the disclosure may be used for agricultural applications, such as delivering to a plant to protect the plant from a pest or pathogen. The water dispersible granules may be applied by diluting the WDG composition in water and spraying onto the plant, plant part, leaf, stem, flower, seed, or soil. In embodiments, the compositions and methods described herein potentiate the efficacy of a pesticidal active ingredient applied simultaneously or separately.

Description

[0001] 128041-5015 GLB-015PC

[0002] 2024-005

[0003] WATER DISPERSIBLE GRANULE FOR AGRICULTURAL APPLICATIONS

[0004] PRIORITY

[0005] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 679,421 filed August 5, 2024, the contents of which are hereby incorporated in their entirety.

[0006] BACKGROUND

[0007] The application of yeast cell walls or components thereof in agriculture may provide benefits for crop health and / or protection from pests and pathogens. While biological mechanisms behind the benefits of yeast cell walls and their components are not well understood, they may offer effective and environmentally friendly solutions for combating crop disease and infestations. However, formulation of yeast cell walls for agricultural application (including large scale applications) is challenging.

[0008] DESCRIPTION OF THE FIGURES

[0009] FIG. 1 shows an exemplary process and systems for producing the water dispersible granule (WDG) compositions of the present disclosure.

[0010] FIG. 2A and FIG. 2B show results of a greenhouse trial for potentiating the action of a pesticide against soybean looper (Chrysodeixis inchidens) on 17-day old soy plants. FIG. 2A shows percent defoliation at Day 11. FIG. 2B shows insect counts on Day 11. ENTRUST biopesticide alone (second bar) showed significant decrease in leaf damage and insect count, compared to untreated control. WDG of the present disclosure potentiated the action of the biopesticide, and performed equivalently to a liquid formulation (bars three and four).

[0011] FIG. 3A and FIG. 3B show further results with WDG compositions prepared with methylated soybean oil or cinnamaldehyde as the organic liquid and used in combination with a commercially available Spinosad-based biopesticide.

[0012] DBl / 161375258.1 1 128041-5015 GLB-015PC 2024-005

[0013] DETAILED DESCRIPTION

[0014] The present disclosure provides a water dispersible granule (WDG) composition comprising yeast particles as a primary granule component (e.g., a substrate) and an organic liquid. The present disclosure further provides methods and processes for making and using the same. The WDG composition of this disclosure comprises yeast particles and an organic liquid and optionally comprises a pesticidal active ingredient, including, for example, a pesticidal RNA (e.g., a pesticidal dsRNA), pesticidal peptide, pesticidal protein, or a pesticidal chemical. In embodiments, the WDG composition (as described further herein) further comprises a dispersing agent and / or a wetting agent, and optionally a filler, and in embodiments does not contain a substantial amount of water.

[0015] The water dispersible granule composition of the disclosure may be used for agricultural applications, such as delivering yeast particles or delivering yeast particles and a pesticidal active ingredient to a plant (for example, a crop) to protect the plant from a pest (for example an insect or mite) or pathogen (for example a fungus, bacteria, or virus). Delivery of the yeast particles and the organic liquid and, optionally a pesticidal active ingredient, to a plant may be accomplished by applying the water dispersible granules to a plant, plant part, leaf, stem, flower, seed, or soil. The water dispersible granules may be applied to a plant, plant part, leaf, stem, flower, seed, or soil by diluting the WDG composition in water and spraying onto the plant, plant part, leaf, stem, flower, seed, or soil. In embodiments, the compositions and methods described herein potentiate the efficacy of a pesticidal active ingredient applied simultaneously (e.g., in the same composition) or separately.

[0016] In embodiments, the water dispersible granules of the present disclosure provide for high loading capacities. For example, yeast particles can be loaded at about 50% or greater, or about 60% or greater, or about 70% or greater, or about 80% or greater, or about 85% or greater by weight of the WDG composition. In some embodiments, the yeast particles comprise from about 40% to about 80% by weight of the WDG composition. In some embodiments, the yeast particles comprise from about 50% to about 70% by weight of the WDG composition. In some embodiments, the yeast particles comprise from about 55% to

[0017] DBl / 161375258.1 2 128041-5015 GLB-015PC 2024-005 about 65% by weight of the WDG composition. In some embodiments, the yeast particles comprise from about 60% to about 65% by weight of the WDG composition. In some embodiments, the yeast particles comprise about 63.5% by weight of the WDG composition. In some embodiments, composition comprises other materials including fillers.

[0018] In embodiments, a pesticidal ingredient, such as a dsRNA active ingredient or a pesticidal protein or chemical pesticide, may be loaded from about 0.1% to about 50%, or from about 0.5% to about 40%, or from about 0.5% or from about 30%, or from about 0.5% to about 20%, or from about 0.5% to about 10% by weight of the WDG composition. In embodiments, the pesticidal ingredient is loaded from about 1% to about 3%, or from about 2% to about 3%, or from about 1% to about 5%, or at about 1%, or at about 2%, or at about 3%, or at about 4%, or at about 5%, or at about 6%, or at about 7%, or at about 8%, or at about 9%, or at about 10%, or at about 15%, or at about 20%, or about 25%, or at about 40%, or at about 45%, or at about 50% by weight of the WDG composition. Comparatively, a liquid (e.g., aqueous) formulation (e.g., for dsRNA active ingredient) would be capable of loading significantly less active ingredient due to the limited capacity of a liquid formulation to maintain a flowable form. Thus, the WDG composition allows for a manufacturer to ship the same amount of yeast particles or the same amounts of yeast particles and pesticidal active ingredient in a composition that weighs significantly less than a liquid formulation with the same amount of the yeast particles or yeast particles and pesticidal active ingredient. When the end user is ready to use the WDG composition, for example, by spraying the composition onto or in the vicinity of a plant, the end user can add water to create a liquid for spraying.

[0019] In embodiments, an advantage of the disclosure is the ability to generate a WDG composition without the need for a biocide (e.g., a preservative) in the composition. A liquid formulation comprising water and yeast particles would be vulnerable to, for example, grampositive and gram-negative rod bacteria which can harm the composition itself and / or harm beneficial organisms that might encounter the product when applied to a plant, such as, for example, the crops onto which the product has been sprayed or organisms that consume such crops, such as humans. Thus a liquid formulation comprising yeast particles and water would typically require the addition of one or more biocides to prevent bacterial growth during

[0020] DBl / 161375258.1 3 128041-5015 GLB-015PC

[0021] 2024-005 storage of the composition. It may be desirable to avoid the use of such biocides, such as where an organic certification is desired. A WDG composition comprising yeast particles without significant amounts of water (e.g., 10% or less by weight) would not typically be vulnerable to bacterial growth. Thus, by eliminating or limiting the use of water in the composition while it is being manufactured, shipped, and stored, until, for example, the end user is ready to apply the composition to a plant, the WDG compositions made according to the present disclosure create significant advantages over liquid (e.g., aqueous) formulations. It is therefore advantageous to reduce residual water in the WDG composition to about 10% or less (e.g., by drying). In embodiments, the WDG composition comprises less than about 10% residual water (by weight), or less than about 5% residual water (by weight), or less than about 3% residual water (by weight) or about 2% residual water (by weight) or less. In embodiments, the WDG composition has about 5% residual water (by weight) or less.

[0022] A still further advantage of the present disclosure is that the addition of organic liquid to the composition reduces the stickiness of the WDG composition, reducing the sticking of the composition to the container in which it is found and reducing the composition sticking to itself (e.g., forming clumps). When water is added to the WDG composition prior to application, the organic liquid desorbs from the yeast, allowing the yeast to break from the organic liquid when diluted and allowing the yeast to be efficiently applied to the plant target. This interaction between the yeast and the organic liquid also allows more efficient delivery of a pesticidal active ingredient if the pesticidal active ingredient is added to the composition. In embodiments, the organic liquid further has adjuvant qualities in the aqueous liquid composition applied to a plant, plant part, or soil, and / or aids in the potentiation of an active ingredient by the yeast particles.

[0023] Thus, in an aspect, the present disclosure provides a method for generating the water dispersible granule composition comprising yeast particles and an organic liquid. The method comprises spraying an organic liquid onto a substrate comprising yeast particles; and converting the sprayed product to a composition comprising water dispersible granules.

[0024] In some embodiments, the yeast particles are selected from the group of yeast cell walls (YCW), components of YCW, and yeast cells (including inactivated yeast cells), and

[0025] DBl / 161375258.1 4 128041-5015 GLB-015PC

[0026] 2024-005 combinations thereof. YCW for use in the invention may include, for example, YCW gathered by autolyzing yeast cells and removing the soluble portion (i.e., removing the yeast extract), and recovering the insoluble portion comprising the YCW. In some embodiments, the yeast particles are commercially available yeast particles. Yeast particles for use in the invention may be further processed by fragmenting the yeast particles through physical (e.g., bead milling) and / or chemical means (e.g., enzymatic digestion) prior to or after being sprayed with the organic liquid. The YCW may comprise all of the cell wall components or may comprise some of the components (e.g., separated by physical and / or chemical means), optionally wherein the components comprise one or more of -glucans (beta-glucans) and mannan-oligosaccharides. In embodiments, the yeast particles comprise P-1,3 glucans, P-1,6 glucans, and / or P-1,3-1,6 glucans, optionally wherein the P-1,3 glucans, P-1,6 glucans, and / or P-1,3-1,6 glucans are branched. In embodiments, the yeast particles comprise intact yeast cell walls or fragmented yeast cell walls. In some embodiments, the fragmented yeast cell walls have been fragmented through a bead milling process. In some embodiments, the yeast particles comprise P-l,6-glucans, P-1, 3 -glucans, mannan-oligosaccharides, mannoproteins, chitin, and / or lipids, optionally wherein the lipids are phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidyl serine (PS), phosphatidic acid (PA), and / or cytidinediphosphate-diacylglycerol (CDP-DAG). Yeast Cell Walls in embodiments are acquired from commercial sources.

[0027] In embodiments, the yeast particles, such as yeast cell walls, may have a starting volume average particle size of about 50 microns to about 100 microns. In embodiments, the volume average particle size of the yeast particles are reduced to a volume average particle size of about 2 to about 10 microns or about 2 to about 5 microns. In embodiments, the water dispersible granules produced by the method of this disclosure have a granule size range (or a mean granule size) of about 0.3 mm to about 10 mm, about 2 mm to about 10 mm, about 0.3 mm to about 6 mm, about 4 mm to about 6 mm, about 3 mm to about 7 mm, or about 0.3 mm to about 8 mm.

[0028] In some embodiments, the method comprises mixing yeast particles, a dispersing agent, a wetting agent, and optionally a filler to create a composition comprising the yeast cell particles (e.g., using a blender), to act as a substrate for the organic liquid and optionally

[0029] DBl / 161375258.1 5 128041-5015 GLB-015PC 2024-005 an active pesticidal ingredient, which can be applied by being sprayed onto the substrate. Figure 1 shows a process and system according to some embodiments.

[0030] In embodiments, the substrate comprising yeast particles are reduced in size using for example a hammer mill and / or an air mill. In embodiments, the volume average particle size of the substrate comprising yeast particles is reduced to a desired size or range through milling to prepare the substrate for the organic liquid (and optionally the pesticidal ingredient), which in embodiments employs air milling and / or a bead milling and optionally further includes liquid nitrogen cooling to improve hammer milling performance and achieve further size reduction of yeast particles. For example, the volume average particle size can be from about 2 to about 10 microns, or from about 2 to about 5 microns. In embodiments, the volume average particle size is about the size of two intact yeast particles or smaller, about 1.5 times the size of a single intact yeast particle or smaller, or about the size of a single intact yeast particle or smaller. In embodiments, the average particle size is reduced to the desired size using a hammer mill followed by an air mill.

[0031] As shown in Figure 1, the organic liquid can be sprayed onto the substrate using a blender and spray bar. In embodiments, the organic liquid is atomized. In embodiments, the substrate sprayed with organic liquid is converted to a wet powder (e.g., with a kneader), and the wet powder is converted to a wet granule (e.g., with an extruder). In embodiments, the wet granules are dried to create a water dispersible granule (e.g., using a fluid bed dryer). Water dispersible granules of the desired size can be recovered by passing through a first mesh (i.e., screen) to remove granules larger than a ceiling value (e.g., 10 mesh to remove particles above about 2 mm, or a 2 mesh to remove particles above about 10 mm), and a second mesh that removes granules below a floor value (e.g., 50 mesh, where the floor value is about 0.3 mm). The water dispersible granules are recovered from the top of the second mesh. Granules removed with the first mesh or the second mesh can be recycled, for example, recycled to a hammer mill for reducing the size of the composition comprising yeast particles.

[0032] In embodiments the organic liquid is selected from a water insoluble organic liquid and a water-soluble organic liquid. In some embodiments, the organic liquid does not

[0033] DBl / 161375258.1 6 128041-5015 GLB-015PC

[0034] 2024-005 dissolve / break up the composition comprising the yeast particles, but rather may reduce the stickiness of the composition. In embodiments, once water is added to the WDG composition before application, the oil separates from the yeast particles while the yeast particles remain in their less-sticky state. In embodiments the organic liquid has adjuvant properties in the aqueous liquid composition applied to a plant, plant part, or soil and / or aids the yeast particles in further potentiating a pesticidal active ingredient.

[0035] In embodiments, the organic liquid does not dampen the granulation ability of the water dispersible granule, does not evaporate upon storage, and does not facilitate binding or degradation of the yeast particles upon storage.

[0036] In some embodiments, the organic liquid is a water insoluble organic liquid, and in embodiments is selected from: disubstituted amides (e.g., commercially available Nodimethyl amides of fatty acids having from six to 22 carbon atoms), aromatic compounds (e.g., Aromatic 100, 150 and 200, D-limonene, Methyl salicylate, cinnamaldehyde), seed oils (e.g., canola, sunflower, grapeseed, cottonseed, safflower, soybean, and com oils, which in some embodiments are ethoxylated or methylated), acetophenone, n-octanol, and Butyl lactate, and combinations thereof. In some embodiments, the organic liquid is a water soluble organic liquid, and in embodiments is selected from propylene carbonate, ethylene carbonate, propylene glycol, ethyl acetate, ethyl lactate, and ethylene glycol, and combinations thereof. In embodiments the organic liquid comprises cinnamaldehyde. In embodiments the organic liquid comprises soybean oil. In embodiments the organic liquid comprises methylated soybean oil. In embodiments, the organic liquid comprises from about 5% to about 25% (by weight) of the final WDG composition. In embodiments, the organic liquid comprises from about 12% to about 20% (by weight) of the final WDG composition. In embodiments, the organic liquid comprises from about 15% to about 18% (by weight) of the final WDG composition.

[0037] The dispersing agent maintains the yeast particles in suspension and prevents or reduces agglomeration or reagglomeration of yeast particles in the mixture when the end user adds water to the mixture to prepare for spraying. The dispersant also functions to aid in returning the yeast particles to suspension after water has been added to the composition

[0038] DBl / 161375258.1 7 128041-5015 GLB-015PC 2024-005 if it is not sprayed for an extended period (e.g., aiding in efficiency of returning the yeast particles into suspension when the mixture is agitated, such as via a pump or stirring).

[0039] In some embodiments, the dispersing agent is selected from naphthalene condensates (e.g., sodium poly[(naphthaleneformaldehyde)sulfonate]), lignosulfonates (e.g, kraft lignin sulfonates), polyacrylates, and phosphate esters, and combinations thereof. In embodiments, the dispersing agent constitutes from about 1% to about 15% by weight of the water dispersible granule composition according to the disclosure. In embodiments, the dispersing agent constitutes from about 1% to about 5% by weight of the water dispersible granule composition according to the disclosure. In embodiments, the dispersing agent constitutes from about 3% to about 7% by weight of the water dispersible granule composition according to the disclosure. In some embodiments, the dispersing agent constitutes from about 4% to about 6% by weight of the water dispersible granule composition according to the disclosure. In some embodiments the dispersing agent constitutes from about 4.5% to about 5.5% by weight of the water dispersible granule composition according to the disclosure. In embodiments, the dispersing agent constitutes about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, or about 15% by weight of the water dispersible granule composition according to the disclosure. In embodiments the dispersing agent comprises one or more kraft lignosulfonates. In some embodiments, the one or more kraft lignosulfonates comprise about 5% by weight of the water dispersible granule according to the disclosure. In some embodiments, the dispersing agent comprises one kraft lignosulfonate with a low degree of sulfonation (e.g., Polyfon® H) and one kraft lignosulfonate with a high degree of sulfonation (e.g., Polyfon® F). In some embodiments the kraft lignosulfonate with a high degree of sulfonation comprises about 4% by weight of the water dispersible granule composition according to the disclosure and the kraft lignosulfonate with a low degree of sulfonation comprises about 1% by weight of the water dispersible granule composition according to the disclosure.

[0040] When water is added to the composition, the wetting agent promotes water uptake onto the surface of the granules, aiding the water in penetrating any cracks in the granules and opening up the granules. Thus the wetting agent in embodiments can reduce the wetting

[0041] DBl / 161375258.1 8 128041-5015 GLB-015PC

[0042] 2024-005 time for the granules and / or reduce the number of inversions of the vessel holding the composition needed to disperse the granules once wetted. In some embodiments, the wetting agent is selected from an alkyl naphthalene sulfonate (e.g., sodium alkyl naphthalene sulfonate powder mixtures), a phosphate ester, a sulphosuccinate, a sodium di octyl sulfosuccinate (e.g., sulfosuccinate products), a nonionic surfactant (e.g., an alcohol ethoxylate), an anionic surfactant (e.g., an alkyl sulfate such as sodium lauryl sulfate and an alkyl benzene sulfonate such as sodium dodecylbenzene sulfonate, and combinations thereof. In some embodiments the wetting agent is a sulphosuccinate. In embodiments, the wetting agent constitutes about 0.1% to about 10% by weight of the water dispersible granule composition according to the disclosure. In embodiments, the wetting agent constitutes about 2.5% or less by weight of the water dispersible granule composition according to the disclosure. In some embodiments, the wetting agent constitutes about 1% to about 5% by weight of the of the water dispersible granule composition according to the disclosure. In some embodiments, the wetting agent constitutes about 1% to about 2.5% by weight of the of the water dispersible granule composition according to the disclosure. In some embodiments, the wetting agent constitutes about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10% by weight of the water dispersible granule composition according to the disclosure.

[0043] In some embodiments, the composition optionally comprises one or more fdlers. In embodiments, the filler is selected from starch, kaolin, bentonite, kieselguhr, Fuller’s earth, attapulgite, diatomaceous earth, bole, loess, talc, chalk, dolomite, limestone, lime, calcium carbonate, powdered magnesia, magnesium oxide, magnesium sulfate, sodium chloride, gypsum, calcium sulfate, pyrophyllite, silicic acid, silicates and silica gels, fertilizers (e.g., ammonium sulfate, ammonium phosphate, ammonium nitrate and urea), natural products of vegetable origin (e.g., grain meals and flours, bark meals, wood meals, nutshell meals and cellulosic powders), and synthetic polymeric materials (e.g., ground or powdered plastics and resins), and combinations thereof. In embodiments, the filler is inert in the WDG. In some embodiments, the filler comprises a fertilizer that does not have a negative effect on the physical performance or stability of the water dispersible granule. In some embodiments the filler constitutes up to about 85% by weight of the water dispersible granule composition

[0044] DBl / 161375258.1 9 128041-5015 GLB-015PC

[0045] 2024-005 according to this disclosure, such as from about 0.1% by weight of the composition to about 85% by weight of the composition, or from about 25% by weight to about 75% by weight of the composition, or from about 25% by weight to about 50% by weight of the composition. In embodiments, the fdler constitutes about 10% or less, or about 5% or less, or about 2.5% or less by weight of the composition, such as from about 0.1% by weight to about 2.5% by weight of the composition, or from about 0.8% by weight to about 1.5% by weight of the composition.

[0046] Some embodiments of the disclosure involve the addition of a pesticidal active ingredient, for example, by spraying it onto the substrate. In embodiments, the pesticidal active ingredient is sprayed onto the substrate prior to, at the same time, or after spraying of the substrate with the organic liquid. The pesticidal active ingredient may be a fungicidal active ingredient, an insecticidal active ingredient, and / or a miticidal active ingredient. In some embodiments, the pesticidal active ingredient is selected from a pesticidal nucleic acid (e.g., a pesticidal dsRNA), a pesticidal protein, and a pesticidal chemical.

[0047] In some embodiments, the pesticide is a ryanodine receptor modulator (IRAC Class 28), an inhibitor of chitin biosynthesis affecting CHS1 (IRAC Class 15), a nicotinic acetylcholine receptor (nACHR) allosteric modulator-site 1 (IRAC Class 5), an nACHR competitive modulator (IRAC Class 4), a sodium channel modulator (IRAC Class 3), an acetylcholinesterase (ACHE) inhibitor (IRAC Class 1), a GABA-gated chloride channel blocker (IRAC Class 2), a glutamic-gated chloride channel (GLUCL) allosteric modulator (IRAC Class 6), a juvenile hormone receptor modulator (IRAC Class 7), a miscellaneous non-specific (multi-site) inhibitor (IRAC Class 8), a chordotonal organ TRPV channel modulator (IRAC Class 9), a mite growth inhibitor affecting CHS1 (IRAC Class 10), an inhibitor of mitochondrial ATP synthase (IRAC Class 12), an uncoupler of oxidative phosphorylation via disruption of the proton gradient (IRAC Class 13), a nicotinic acetylcholine receptor (nACHR) channel blocker (IRAC Class 14), an inhibitor of chitin biosynthesis type 1 (IRAC Class 16), a molting disruptor dipteran (IRAC Class 17), an ecdysone receptor agonist (IRAC Class 18), an octopamine receptor agonist (IRAC Class 19), a mitochondrial complex III electron transport inhibitor QO site (IRAC Class 20), a mitochondrial complex I electron transport inhibitor (IRAC Class 21), a voltage-dependent

[0048] DBl / 161375258.1 10 128041-5015 GLB-015PC 2024-005 sodium channel blocker (IRAC Class 22), an inhibitor of acetyl-CoA carboxylase (IRAC Class 23), a mitochondrial complex IV electron transport inhibitor (IRAC Class 24), a mitochondrial transport inhibitor (IRAC Class 25), a chordotonal organ nicotinamidase inhibitor (IRAC Class 29), a GABA-gated chloride channel allosteric modulator (IRAC Class 30), a nicotinic acetylcholine receptor (NACHR) allosteric modulator - site II (IRAC Class 32), a calcium-activated potassium channel (KCa2) modulator (IRAC Class 33), a mitochondrial complex III electron transport inhibitor QI site (IRAC Class 34), or a chordotonal organ modulator - undefined target site (IRAC Class 36), or combinations thereof. In some embodiments, the chemical pesticide is selected from chlorantraniliprole, novaluron, spinosad, acetamiprid, cypermethrin, diflubenzuron, and bifenthrin.

[0049] In some embodiments the pesticidal active ingredient is a pesticidal RNA that selectively causes RNA interference resulting in control of the targeted pest. Laboratory and field studies have demonstrated that oral delivery of RNA molecules whose mode of action is through the RNAi process (e.g., double-stranded RNA (dsRNA)) are effective for many insect and mite species and hence, topical dsRNA is considered a suitable form of delivery. See, US 11,185,079 (titled “Control of Coleopteran Insects”), US 11,142,768 (titled, “Control of Insect Infestation”), US 2023 / 0345947 Al (titled “RNA-based Control of Lepidopteran Pests”), WO 2020 / 081487 Al (titled “Control of Coleopteran Insect Infestation”), US 2022 / 0061335 Al (titled “Control of Insect Infestation”), US 10,378,012 (titled “Compositions and Methods for Controlling Insect Pests”), and US 10,975,387 (titled “Compositions and Methods for Controlling Insect Pests”), all of which are hereby incorporated by reference in their entireties. Moreover, essential genes can be silenced for the control of pathogens via RNA interference without adversely affecting non-target species, allowing growers to target pests more precisely and in an environmentally friendly manner compared to conventional agrochemicals (Cagliari et al., Front. Plant Set. 2019). RNAi based technologies have also shown effectiveness in controlling fungal pathogens as well. See, U.S. 12,077,763 (titled “RNA-based Control of Powdery Mildew”), US 2023 / 0189818 Al (titled “RNA-Based Control of Botrytis”), and WO 2023 / 196934 Al (titled “RNA-Based Control of Production of Deoxynivalenol by Fusarium”), all of which are hereby incorporated by reference. RNAi molecules targeting insect or fungal genes may

[0050] DBl / 161375258.1 11 128041-5015 GLB-015PC

[0051] 2024-005 be produced by any method known in the art, including for example, according to cell-free production methods described, for example, in U.S. Patent Nos. 10,954,541 (entitled “Cell- free production of ribonucleic acid”) and 10,858,385 (entitled “Methods and compositions for nucleoside triphosphate and ribonucleic acid production”), each of which is hereby incorporated by reference.

[0052] In embodiments, the present disclosure provides a process for creating a WDG (according to embodiments as described above), using the system and / or following steps depicted in FIG 1. In embodiments, the process comprises the following steps, with reference to FIG. 1.

[0053] 1. The yeast particle, dispersing agent, wetting agent, and optionally fillers are added into a blender, and mixed until they become a homogeneous composition.

[0054] 2. The blender is discharged through a hammer mill, where the average particle size of the composition is reduced (as described herein).

[0055] 3. The product of step 2 is discharged through an air mill or a bead mill. An air mill can be efficient for use on a dry product and therefore is preferable in some embodiments. The air milling process may be varied to impact the size distribution of the yeast particles: (a) traditional air-milling techniques can be used to deagglomerate the yeast particles to the primary particle size of about 2 to about 5 microns; or (b) high pressure (about 120 PSI), co-milling with dry ice, or steam, can be used to rupture the yeast particles. Rupture of yeast particles can be observed by particle size measurements, as increased fines less than the primary size of the yeast particles.

[0056] 4. The organic liquid is added to the product of step 3 using a blender with spray bar, and the organic liquid is absorbed onto the yeast surface.

[0057] 5. The product of step 4 is placed into a kneader, where water is added to make a dough that can be extruded. Optimal kneader efficiency is expected from a high shear, such as with a SCHUGI FLEXOMIX.

[0058] DBl / 161375258.1 12 128041-5015 GLB-015PC

[0059] 2024-005

[0060] 6. The kneaded material from step 5 is placed in an extruder, optionally a basket extruder or a radial extruder.

[0061] 7. The wet extruded granules of step 6 are dried, such as by being dropped into a fluid bed dryer and dried to specification moisture content (as already described).

[0062] 8-9. The dried product of step 7 may be sieved to the desired particle size through screens (including as already described). Overs (granules bigger than the desired size) and fines (granules smaller than the desired size) may be removed from the WDG composition. In step 8, the dried product of step 7 is passed through a screen that allows passage of only granules under the desired maximum particle size. In step 9, the product that passes through the screen in step 8 is then passed through a screen that allows through only products below the minimum desired specification to pass through.

[0063] 10. The product remaining on top of the screen in step 9 is gathered as the WDG composition in step 10.

[0064] 11. Products that do not pass through the screen of step 8 (i,e., the overs) and products that do pass through the screen of step 9 (i.e., the unders), are collected in step 11 and may be discarded or recycled back into the system to be reprocessed, optionally starting at the hammer mill application at step 2.

[0065] In some embodiments, a pesticidal active ingredient is added (e.g., applied by spraying) to the substrate comprising yeast particles, dispersing agent, wetting agent, and organic liquid, and optionally a filler (e.g., according to the embodiments already described). In some embodiments, prior to its application to the substrate, the pesticidal active ingredient may be prepared in a formulation selected from an emulsifiable concentrate (EC), a dispersible concentrate (DC), a microemulsion concentrate (ME), a suspension concentrate (SC) and a soluble concentrate (SL). In embodiments, the pesticidal active ingredient is added to the substrate comprising yeast particles by being sprayed onto the surface of such substrate before, after, or at the same time as the organic liquid is sprayed onto the surface of such substrate.

[0066] DBl / 161375258.1 13 128041-5015 GLB-015PC 2024-005

[0067] In embodiments in which an EC or DC formulation of a pesticidal active ingredient is applied to the substrate, the pesticidal active ingredient comprises about 5% to about 30% by weight of the EC or DC formulation; a solvent comprises about 50% to about 80% by weight of the EC or DC formulation; and an ethoxylated emulsifier comprises about 1% to about 10% of the EC or DC formulation. In some embodiments the solvent is selected from Aromatic 200, d-limonene, acetophenone, ethyl acetate, octanol, butyl lactate, methyl salicylate, propylene carbonate, ethylene carbonate, propylene glycol, ethylene glycol, and combinations thereof. Other suitable solvents for a particular pesticidal active ingredient can be selected as known in the art, and for use in any of an EC, DC, ME, SC, or SL. See, Pesticide Formulations and Application Systems: Eighth Volume, By DA Hovde and GB Beestman, ASTM International, 1989, Chapter An Overview — Solvents for Agricultural Chemicals. Generally, such solvents include the known hydrocarbon solvents, aromatic solvents, paraffinic solvents, aliphatic solvents, petroleum oils, oxygenated solvents, and green solvents (sustainable solvents), bio-based solvents, and water.

[0068] In embodiments, the formulation of a pesticidal active ingredient comprises an emulsifier. In embodiments, the emulsifier is selected from linear and branched alcohol ethoxylates, castor oil ethoxylates, polysorbate ethoxylates, nonylphenol ethoxylates, tri styrylphenol ethoxylates, coco and tallowamine ethoxylates, tall oil fatty acid ethoxylates, tridecyl alcohol ethoxylates, alkoxylated amides, triblock copolymers (e.g., poloxamers), random copolymers (e.g., butyl based), poloxamers, reverse poloxamers, and acrylic copolymers, and combinations thereof.

[0069] In embodiments, the EC or DC formulation of a pesticidal active ingredient is prepared by adding the pesticidal active ingredient to a solvent and dissolving at about 50° C and then adding and dissolving the emulsifier. The resulting formulation may then be sprayed onto the surface of the substrate comprising yeast particle, dispersing agent, wetting agent, and optionally filler, at the same time, or before, or after the organic liquid is sprayed onto the surface of such substrate (e.g., in Step 4 of Figure 1).

[0070] In embodiments in which an SC formulation of a pesticidal active ingredient is applied to the substrate, the pesticidal active ingredient comprises about 5% to about 60%

[0071] DBl / 161375258.1 14 128041-5015 GLB-015PC

[0072] 2024-005 by weight of the SC formulation, a polymeric dispersing agent comprises about 1% to about 10% of by weight of the SC formulation, an ethoxylated dispersing agent comprises about 1% to about 10% by weight of the SC formulation, an antifoaming agent comprises about 0.01% to about 0.1% by weight of the SC formulation, a rheology modifier comprises about 0.01% to about 0.1% by weight of the SC formulation and a carrier comprises about 30% to about 80% by weight of the SC formulation. In some embodiments, the antifoaming agent is a trisiloxane antifoaming agent. In some embodiments, the rheology modifier is xanthan gum. In some embodiments the carrier is water.

[0073] In some embodiments, the SC formulation of a pesticidal active ingredient is prepared by adding the polymeric dispersing agent and ethoxylated dispersing agent to water and dissolving; adding the antifoaming agent and dissolving; adding the pesticidal active ingredient and mixing under bulking agitation until dispersed; reducing the particle size of the pesticidal active ingredient to a volume average particle size of less than about 50 microns (optionally using a rotor stator high shear mixer to reduce particle size); bead milling the resulting formulation to a volume average particle size of about 3 to about 10 microns; and adding the rheology modifier and mixing until fully swelled. The SC formulation of a pesticidal active ingredient may then be sprayed onto the surface of the substrate comprising yeast cell wall, dispersing agent, wetting agent, and optionally filler, at the same time, or before, or after the organic liquid is sprayed onto the surface of such substrate (e.g., in Step 4 of Figure 1).

[0074] In other aspects, the present disclosure provides a water dispersible granule composition comprising yeast particles and an organic liquid coating (and optionally the other components as described herein for various embodiments), and optionally a pesticidal active ingredient (as described herein), wherein the yeast particles have a volume average particle size of about 2 to about 10 microns; and wherein the water dispersible granules have an granule sizes from about 0.3 to about 10 mm. The water dispersible granule may be produced by a method described herein. In embodiments, the composition further comprises a dispersing agent, a wetting agent, and optionally a filler, each as already described. In embodiments, the WDG composition contains a low amount of water. In embodiments, the

[0075] DBl / 161375258.1 15 128041-5015 GLB-015PC 2024-005

[0076] WDG composition comprises less than about 20 % water (by weight), such as less than about 20% (by weight), less than about 10% (by weight), or less than about 5% (by weight).

[0077] In embodiments, the WDG composition of this disclosure is diluted with water, and applied (e.g., by spraying) to plants (e.g., crops) as described, including plants that are infested or at risk of infestation by one or more pests or pathogens. In embodiments, the WDG composition is diluted with water at a ratio of at least 10:1 (water: WDG), or at least 100: 1, or at least 200: 1, or at least 500: 1, or at least 1000: 1, or at least 2000: 1. In embodiments, the WDG composition is diluted at about 2000: 1 or less, or about 1000: 1 or less, such as from about 100: 1 to about 300: 1 (e.g., about 200: 1).

[0078] In some embodiments, the plant pest is an insect belonging to the order Lepidoptera, Coleoptera, Hemiplera. Diptera, or Acari. In some embodiments, the plant pest is resistant to Bacillus thuringiensis (Bt) or a Bt protein. In embodiments, the plant is infected or at risk of infection by a fungus.

[0079] In embodiments, the plant pest belonging to the order Lepidoptera is a species from a family selected from Nymphalidae (brush-footed butterflies), Danaidae (milkweed butterflies), Pieridae (whites and sulfurs) Papilionidae (swallowtails), Lycaenidae (blues, coppers, and hairstreaks), Hesperiidae (skippers), Tineidae (clothes moths), Sesiidae (clearwing moths), Pyralidae (snout moths), Lasiocampidae (lappet moths), Satumiidae (giant silk moths), Sphingidae (hawk moths), Arctiidae (tiger moths), Lymantriidae (tussock moths), Noctuidae (loopers, owlet moths, and underwings), and Plutellidae (diamond back moths). In embodiments, the Lepidopteran insect species is Plutella spp. (e.g., Plutella xylostella). In embodiments, the Lepidopteran insect species is Spodoptera spp. (e.g., Spodoptera frugiperda, Spodoptera exigua, Spodoptera litura, Spodoptera litoralis). In embodiments, the Lepidopteran insect species is Chrysodeixis spp. (e.g., Chyrysodeixis includens or Chyrysodeixis acuta). In embodiments, the Lepidopteran insect species is Helicoverpa spp. (e.g., Helicoverpa armigera or Helicoverpa zea). In embodiments, the Lepidopteran insect species is Plutella spp. (e.g., Plutella xylostella). In embodiments, the Lepidopteran insect species is Spodoptera spp. (e.g., Spodoptera frugiperda). In embodiments, the Lepidopteran insect species is Cydia spp. (e.g., Cydia pomonella). In

[0080] DBl / 161375258.1 16 128041-5015 GLB-015PC 2024-005 embodiments, the Lepidopteran insect species is Trichoplusia spp. (e.g., Trichoplusia ni). In embodiments, the Lepidopteran insect species is Pieris spp. (e.g., Pieris rapae). In embodiments, the Lepidopteran insect species is Tuta spp. (e.g., Tuta absolute). In embodiments, the Lepidopteran insect species is Paramyelois spp. (Paramyelois transitella). In embodiments, the Lepidopteran insect species is Lobesia spp. (e.g., Lobesia botrana).

[0081] In some embodiments, the insect belongs to the order Coleoplera, including species selected from Leptinotarsa spp., Phyllotreta spp., Cerotoma spp., Diabrotica spp., Tribolium spp., Anthonomus spp. and Alticini spp. In embodiments, the Colepotera insect species is Phyllotreta spp. (e.g., Phylotreta Cruciferae or Phylotreta striolata). In embodiments, the Colepotera insect species is Psylliodes spp. (e.g., Psylliodes chrysocephala).

[0082] In embodiments, the composition is applied to a plant selected from Solanaceae plants, Brassicaceae plants, Poaceae plants, Cucurbitaceae plants, Fabaceae plants, Fagaceae plants, Asteraceae plants, Amaryllidaceae plants, Umbelliferae plants, Apiaceae plants, Amranthaceae plants, and Malvaceae plants. For example, in embodiments, the composition is applied to corn, soy, or cotton plants.

[0083] In embodiments the composition is applied to a specialty crop, including but not limited to fruits, vegetables, tree nuts, dried fruits, and horticulture and nursery crops.

[0084] In embodiments, the composition to be sprayed onto a plant, plant part, or soil is applied at a use rate of about 10 L / Ha or more, or from about 10 L / Ha and up to 2,000 L / Ha. In embodiments the composition to be sprayed onto a plant, plant part, or soil is applied at a use rate of about 100 L / Ha to about 500 L / Ha, or about 200 L / Ha, or about 250 L / Ha, or about 300 L / Ha.

[0085] As used herein, the term “about” means ±10% of a reference value, unless the context requires otherwise.

[0086] EXAMPLES

[0087] Example 1 - Preparation of Water Dispersible Granules with Yeast Cell Walls and Methylated Soybean Oil.

[0088] DBl / 161375258.1 17 128041-5015 GLB-015PC 2024-005

[0089] Commercially available yeast cell walls were combined with a dispersing agent (POLYFON F and POLYFON H from Ingevity), wetting agent (AEROSOL OT-B from Syensqo), and filler (commodity Ammonium Sulfate) into a beaker. The resulting mixture was homogenized by stirring with scoopula for 30 seconds and then the mixture was passed through an IKA Mills MF 10 basic Microfme grinder device equipped with MF 10.2 Impact grinding head in order to de-aggregate yeast particles and further homogenize the mixture. Once the mixture was milled, the mixture was passed through a STURDEVANT 2” Microionizer air mill in order to further reduce the particle size. Particle size was verified using MICROTRAC S3500 particle size analyzer until particles reach 2-10 microns in size. Once desired particle size was reached, the milled material is transferred into CUISINART Elite Collection 2.0 14 Cup Food Processor. While the food processor runs, STEPOSOL ME (Stepan) organic solvent was added (a commercially available methylated soybean oil or methyl soyate) onto the substrate using VEEJET stainless steel flat fan nozzle. The organic solvent was applied while under continuous mixing in food processor. The texture of the mixture will go from dry powder to oily powder as the organic solvent is applied. Once the organic solvent is fully added, mixing of the substrate was continued for homogeneity. Once the mixture is homogeneous and no dry clumps or patches remain in the mixture, kneading water is added at approximately 30% by weight of the oily mixture in the food processer while under mixing conditions. The material will go from oily powder to wet, sticky texture. The wetted substrate was transferred to a benchtop basket extruder and the material extruded into wet granules. Wet granules were collected and transferred to SHERWOOD Model 501 Fluid Bed Dryer, which was run at 70° C for 12 minutes to evaporate water from the granules. The granules transitioned from wet to dry and turned from dark brown to lighter brown as they dried. Once dried, the residual water content of produced granules is measured using loss on drying (LOD) method.

[0090] Physical properties of the granules were determined using CIPAC methods as shown in Table 1:

[0091] Table 1 : WDG Evaluation

[0092] DBl / 161375258.1 18 128041-5015 GLB-015PC 2024-005

[0093] Example 2 - Preparation of Water Dispersible Granule with Yeast Cell Walls, Methylated Soybean Oil, and Spinosad

[0094] A Spinosad containing yeast cell wall WDG was prepared as follows. Commercially available yeast cell walls were combined with a dispersing agent (POLYFON F and POLYFON H from Ingevity), a wetting agent (AEROSOL OT-B by Syensqo), and filler (commodity ammonium sulfate) in a beaker. The resulting mixture was homogenized by stirring with scoopula for 30 seconds and then passing the mixture through an IKA Mills MF 10 basic Microfine grinder device equipped with MF 10.2 Impact grinding head in order to de-aggregate yeast particles and further homogenize the mixture. Once the mixture was milled, the mixture was passed through a STURDEVANT 2” Microionizer air mill to further reduce the particle size. Particle size is verified using MICROTRAC S3500 particle size analyzer until particles reach 2-10 microns in size. Once desired particle size was reached, the milled material was transferred to a CUISINART Elite Collection 2.0 14 Cup Food Processor. While the food processor runs, methylated soybean oil (STEPOSOL ME) and commercially available Spinosad technical grade active ingredient (TGAI) dissolved in commodity acetophenone organic solvent was added to the substrate using VEEJET stainless steel flat fan nozzle. The organic solvent and Spinosad solutions were added while under continuous mixing in the food processor. The texture transformed from dry powder to oily powder as the organic solvent is applied. Once organic solvent is fully added, the mixing was continued for homogeneity. Once the mixture is homogeneous and no dry clumps or

[0095] DBl / 161375258.1 19 128041-5015 GLB-015PC

[0096] 2024-005 patches remain in the mixture, kneading water at approximately 30% by weight of the oily mixture was added to the food processer while under mixing conditions. The material was transformed from oily powder to wet, sticky texture. The wetted substance was transferred to a benchtop basket extruder and the material extruded into wet granules. Wet granules were collected and transferred to SHERWOOD Model 501 Fluid Bed Dryer, run at 70° C for 12 minutes in order to evaporate water from the granules. The granules transitioned from wet to dry and turned from dark brown to lighter brown as they dry. Once dried, the residual water content was determined using loss on drying (LOD) method.

[0097] CIPAC assays were performed as described in Example 1. Spinosad quantification analysis was conducted to confirm the desired composition was attained.

[0098] Example 3 - Performance of WPG Compositions

[0099] Yeast cell wall WDG compositions prepared according to methods of this disclosure (e.g., WDG compositions prepared essentially as described in Example 1) perform equivalently to liquid compositions in improving the performance of pesticidal active ingredients, as demonstrated in Figures 2A and 2B.

[0100] A yeast cell wall WDG composition (PF-X2-I-005) was prepared in a similar manner to that described in Example 1 , comprising about 64% Yeast Cell Walls and 24% methylated soybean oil. Additionally a liquid formulation of yeast cell walls and methylated soybean oil was also prepared, comprising about 25% fractured yeast cell walls and 9.5% methylated soybean oil (PF -XI -021).

[0101] Compositions were tested in a greenhouse trial of efficacy against soybean looper (Chrysodeixis includens) 1stinstar larvae, on 17-day old soy plants with two plants per cage and 9 larvae per plant. A preventative application of (1), (2), or (3) was made on day one, using a handheld CO2 pressurized spray with a volume of 50 gallons per acre with tap water as the carrier: (1) ENTRUST, a commercially available Spinsosad-derived biopesticide, at 6.3 gA / ha; (2) PF-X1 -1-021 (a suspoemulsion formulation of yeast cell walls and methylated soybean oil) at 2 L / ha plus ENTRUST at 6.3 gA / ha; and (3) PF-X2-I-005 (WDG yeast cell wall and methylated soybean oil) at 700 gA / ha plus ENTRUST at 6.3 gA / ha. Plant

[0102] DBl / 161375258.1 20 128041-5015 GLB-015PC

[0103] 2024-005 defoliation was assessed on day 7 and 11 and insect counts determined on day 11. Results are shown in Figures 2A (% defoliation at day 11) and 2B (insect count per cage). ENTRUST alone (second bar) showed significant decrease in leaf damage and insect count, compared to untreated control (first bar). The groups combining ENTRUST plus the SE formulation PF-X 1-1-021 (third bar) and WDG formulation PF-X2-I-005 (fourth bar) performed equivalently to one another and demonstrated a significant improvement vs. ENTRUST alone.

[0104] Example 4 - Performance of WDG Compositions

[0105] Yeast cell wall WDG compositions prepared according to methods of this disclosure (e.g., WDG compositions prepared essentially as described in Examples 1 and 2) improved the performance of pesticidal active ingredients, as demonstrated in Figures 3A and 3B, including WDG compositions prepared with methylated soybean oil or cinnamaldehyde as the organic liquid and used in combination with a commercially available Spinosad-based biopesticide.

[0106] Yeast cell wall WDG composition (PF-X2-I-) was prepared in a similar manner to that described in Example 1 with PX-X2-I-005 comprising about 64% Yeast Cell Walls and 24% methylated soybean oil. Yeast cell wall WDG composition (PF-X2-I-006) was prepared in a similar manner to that described in Example 1 (except for the use of cinnamaldehyde as the organic liquid in place of methylated soybean oil) comprising about 64% yeast cell walls and 24% cinnamaldehyde. A yeast cell wall, methylated soybean oil, and Spinosad WDG composition (PF-X3-I-009) was prepared in a similar manner to that described in Example 2, comprising about 4% Spinosad, 65% yeast, and 16% methylated soybean oil. Additionally a liquid formulation of yeast cell walls and methylated soybean oil was also prepared, comprising about 25% fractured yeast cell walls and 9.5% methylated soybean oil (PF-X1- 1-021).

[0107] Compositions were tested in a greenhouse trial of efficacy against soybean looper (Chrysodeixis includens) on old soy plants with two plants per cage and five 4thinstar larvae per plant (10 per cage). A preventative application of (1), (2), or (3) was made on day one with water as the carrier: (1) ESTERO, a commercially available Spinosad-derived

[0108] DBl / 161375258.1 21 128041-5015 GLB-015PC

[0109] 2024-005 biopesticide, at 1.88 gA / ha; (2) PF-X1 -1-021 (a suspoemulsion formulation of yeast cell walls and methylated soybean oil at 2 L / ha, plus ESTERO at 1.88 gA / ha; (3) PF-X2-I-005 (WDG yeast cell wall and methylated soybean oil) at 700 gA / ha, plus ESTERO at 1.88 gA / ha; (4) PF-X2-I-006 (yeast cell wall and cinnamaldehyde WDG) at 700 gA / ha, plus ESTERO at 1.88 gA / ha; and (5) PF-X3-I-009 (yeast cell wall, methylated soybean oil, and

[0110] Spinosad WDG) at 700 gA / ha. Plant defoliation and insect counts were determined at the end of the trial. Results are shown in Figures 3A (% defoliation at end of trial) and 3B (insect count per cage). ESTERO alone (second bar) showed significant decrease in insect count and a numerical reduction in defoliation, compared to untreated control (first bar). The test groups of ESTERO plus the SE formulation PF-X1 -1-021 (third bar), ESTERO plus WDG yeast cell wall / methylated soybean oil formulation PF-X2-I-005 (fourth bar), ESTERO plus WDG yeast cell wall / cinnamaldehyde formulation PF-X2-I-006 (fifth bar), and the WDG comprising yeast cell wall, methylated soybean oil and Spinosad, all showed significant improvement compared to ESTERO alone and compared to the untreated control.

[0111] DB1 / 161375258.1 22

Claims

128041-5015 GLB-015PC 2024-005CLAIMS:

1. A method for generating a composition comprising water dispersible granules comprising yeast particles, the method comprising:(a) spraying an organic liquid onto a substrate comprising yeast particles; and(b) converting a product of step (a) to a composition comprising water dispersible granules.

2. The method of claim 1, wherein the yeast particles are selected from yeast cell walls and yeast cell wall components, optionally wherein the yeast particles are fragmented yeast cell walls, and optionally wherein the yeast cell walls are fragmented by a method selected from bead milling and enzymatic digestion.

3. The method of claim 1, wherein the yeast particles comprise yeast cell walls.

4. The method of claim 1, wherein the yeast particles comprise one or more yeast cell wall components selected from 0-glucans (beta-glucans) and mannan-oligosaccharides, P- 1,3 glucans, P-1,6 glucans, and / or P-1,3-1,6 glucans, branched P-1,3 glucans, branched P- 1,6 glucans, branched P-1,3-1,6 glucans, mannoproteins, chitin, lipids, phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidic acid (PA), cytidinediphosphate-diacylglycerol (CDP-DAG), or combinations thereof.

5. The method of any one of claims 1 to 4, wherein the yeast particles are present in the WDG at 40% or greater by weight.

6. The method of any one of claims 1 to 5, wherein the yeast particles are present from about 40% to about 80% of the composition by weight.

7. The method of any one of claims 1 to 6, wherein the yeast particles comprise from about 60% to about 65% by weight of the water dispersible granules.

8. The method of claim 7, wherein the yeast particles comprise about 63.5% by weight of the water dispersible granulesDBl / 161375258.1 23128041-5015 GLB-015PC2024-0059. The method of any one of claims 1 to 8, wherein the yeast particles have a volume average particle size of about 2 to about 10 microns; and / or the water dispersible granules have a granule sizes from about 0.3 to about 10 mm, or about 2 mm to about 10 mm, or about 0.3 mm to about 6 mm, about 4 mm to about 6 mm, about 3 mm to about 7 mm, or about 0.3 mm to about 8 mm.

10. The method of any one of claims 1 to 9, where the substrate is prepared by mixing the yeast particles, a dispersing agent, a wetting agent, and optionally a filler, and optionally wherein the mixing is conducted in a blender.

11. The method of any one of claims 1 to 10, wherein the substrate is reduced to a volume average particle size of from about 2 to about 10 microns, or about 2 to about 5 microns; optionally wherein the substrate is reduced from a volume average particle size of about 50 microns to about 100 microns, down to a volume average particle size of about 2 to about 10 microns, or about 2 to about 5 microns.

12. The method of claim 11, wherein the volume average particle size is reduced with a hammer mill, and an air mill and / or a bead mill, optionally wherein the hammer mill utilizes liquid nitrogen.

13. The method of claim 12, the volume average particle size is reduced to about 2 to about 5 microns using a hammer mill followed by an air mill.

14. The method of any one of claims 1 to 13, wherein the spraying is conducted with a blender and spray bar, and optionally wherein the organic liquid is atomized.

15. The method of claim 14, further comprising converting the substrate sprayed with the organic liquid into a wet powder, optionally using a kneader.

16. The method of claim 15, further comprising converting the wet powder into a wet granule, optionally using an extruder.

17. The method of claim 16, further comprising converting the wet granule into a water dispersible granule, optionally using a fluid bed dryer, optionally wherein the water dispersible granule comprises about 10% or less residual water by weight or less than aboutDBl / 161375258.1 24128041-5015 GLB-015PC2024-0057.5% residual water by weight, or about 5% or less residual water by weight, or about 3% or less residual water by weight, or about 2% or less residual water by weight.

18. The method of any one of claims 1 to 17, further comprising: passing the water dispersible granules through a first mesh to remove granules larger than a ceiling value for a desired granule size range and recovering granules smaller than the ceiling value, optionally wherein the ceiling value is about 10 mm; passing the recovered water dispersible granules though a second mesh that removes granules below a floor value for a desired granule size range and recovering granules larger than the floor value, optionally wherein the floor value is about 0.3 mm; and collecting the water dispersible granules from the top of the second mesh.

19. The method of claim 18, further comprising: recycling the granules removed with the first mesh or the second mesh, optionally wherein the recycled granules are used to prepare the substrate, and is optionally recycled to a hammer mill for reducing a volume average size of the substrate.

20. The method of any one of claims 1 to 19, wherein the organic liquid is selected from a water insoluble organic liquid and a water-soluble organic liquid, optionally wherein the organic liquid has adjuvant qualities, optionally wherein the organic liquid potentiates and / or aids in the potentiation of an active ingredient by the yeast particles.

21. The method of claim 20, wherein the organic liquid is a water insoluble organic liquid, and the water insoluble organic liquid is selected from disubstituted amides, aromatic compounds, seed oil(s), and corn oil(s), acetophenone, n-octanol, and Butyl lactate, and combinations thereof.

22. The method of claim 20 or 21, wherein the water insoluble organic liquid is an ethoxylated or methylated seed or com oil, and optionally comprises methylated soybean oil.DBl / 161375258.1 25128041-5015 GLB-015PC2024-00523. The method of claim 21, wherein the organic liquid is a water soluble organic liquid, which is selected from propylene carbonate, ethylene carbonate, propylene glycol, ethyl acetate, ethyl lactate and ethylene glycol, and combinations thereof.

24. The method of any of claims 20 to 23, wherein the organic liquid comprises from about 5% to about 25% (by weight) of the final water dispersible granule, or from about 12% to about 20% (by weight) of the water dispersible granule, or from about 15% to about 18% (by weight) of the water dispersible granule.

25. The method of any one of claims 10 to 24, wherein the dispersing agent is selected from naphthalene condensates, lignosulfonates, polyacrylates, and phosphate esters, and mixtures thereof.

26. The method of claim 25, wherein the dispersing agent is selected from naphthalene condensates, a kraft lignin sulfonate, and mixtures thereof, optionally wherein the dispersing agent comprises one or more kraft lignosulfonates, optionally wherein the one or more lignosulfonates comprise one lignosulfonate with a low degree of sulfonation and one kraft lignosulfonate with a high degree of sulfonation, optionally wherein the kraft lignosulfonate with a high degree of sulfonation comprises about 4% by weight of the water dispersible and the kraft lignosulfonate with a low degree of sulfonation comprises about 1% by weight of the water dispersible granule.

27. The method of any of claims 10 to 26, wherein the dispersing agent constitutes from about 1% to about 15% by weight of the water dispersible granule or from about 1% to about 5% by weight of the water dispersible granule, from about 3% to about 7% by weight of the water dispersible granule, or from about 4% to about 6% by weight of the water dispersible granule, or from about 4.5% to about 5.5% by weight of the water dispersible granule, or about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, or about 15% by weight of the water dispersible granule.

28. The method of any of claims 10 to 27, wherein the wetting agent is selected from the group of an alkyl naphthalene sulfonate, a phosphate ester, a sulphosuccinate, a sodiumDBl / 161375258.1 26128041-5015 GLB-015PC2024-005 dioctylsulfosuccinate, a nonionic surfactant, an anionic surfactant, an alkyl benzene sulfonate and mixtures thereof.

29. The method of claim 28, wherein the wetting agent is selected from a sodium alkyl naphthalene sulfonate powder mixture, a sulfosuccinate, an alcohol ethoxylate, an alkyl sulfate, sodium lauryl sulfate, alkyl benzene sulfonate, sodium dodecylbenzene sulfonate, and mixtures thereof.

30. The method of any one of claims 10 to 29, wherein the wetting agent constitutes about 0.1% to about 10% by weight of the water dispersible granule, or about 2.5% or less by weight of the water dispersible granule or about 1% to about 5% by weight of the of the water dispersible granule or about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10% by weight of the water dispersible granule composition.

31. The method of any one of claims 10 to 30, wherein the filler is selected from kaolin, bentonite, kieselguhr, Fuller’s earth, attapulgite, diatomaceous earth, bole, loess, talc, chalk, dolomite, limestone, lime, calcium carbonate, powdered magnesia, magnesium oxide, magnesium sulfate, sodium chloride, gypsum, calcium sulfate, pyrophyllite, silicic acid, silicates and silica gels, fertilizers, natural products of vegetable origin, and synthetic polymeric materials, and mixtures thereof.

32. The method of claim 31, wherein the filler is selected from ammonium sulfate, ammonium phosphate, ammonium nitrate, urea, grain meals and flours, bark meals, wood meals, nutshell meals and cellulosic powders, a ground plastic, a powdered plastic, a ground resin, a powdered resin, and mixtures thereof.

33. The method of claim 31 or 32, wherein the filler is inert in the water dispersible granule.

34. The method of any of claims 31 to 33, wherein the filler is a fertilizer, optionally wherein the fertilizer does not have a negative effect on the physical stability of the water dispersible granule.DBl / 161375258.1 T1128041-5015 GLB-015PC2024-00535. The method of any one of claims 10 to 34, wherein the filler constitutes up to about 85% by weight of the composition or from about 0.1% by weight of the composition to about 85% by weight of the composition, or from about 25% by weight to about 75% by weight of the composition, or from about 25% by weight to about 50% by weight of the composition, or from 0.1% by weight to about 2.5% by weight of the composition, or from about 0.8% by weight to about 1.5% by weight of the composition, or about 2.5% or less by weight of the composition..

36. The method of any one of claims 1 to 35, further comprising the step of: spraying a pesticidal active ingredient onto the substrate, either prior to, at the same time as, or after spraying of the substrate with the organic liquid.

37. The method of claim 36, wherein the pesticidal active ingredient is a pesticidal RNA, a pesticidal peptide, a pesticidal protein, or a pesticidal chemical, optionally wherein the pesticidal active ingredient is loaded at about 0.1% to about 50% by weight of the composition.

38. The method of claim 36 or 37, wherein the pesticidal active ingredient is a formulation selected from the group of emulsifiable concentrate (EC), a dispersible concentrate (DC), a microemulsion concentrate (ME), a suspension concentrate (SC) and a soluble concentrate (SL), and combinations thereof.

39. The method of claim 38, wherein the pesticidal active ingredient is formulated as an emulsifiable concentrate (EC).

40. The method of claim 39, wherein the EC comprises the pesticidal active ingredient, a solvent, and an ethoxylated emulsifier.

41. The method of claim 39 or 40, wherein the pesticidal active ingredient comprises about 5% to about 30% by weight of the EC, the solvent comprises about 50% to about 80% by weight of the EC, and the ethoxylated emulsifier comprises about 1% to about 10% by weight of the EC.

42. The method of any one of claims 39 to 41, wherein the solvent is selected from the group of Aromatic 200, d-limonene, acetophenone, ethyl acetate, octanol, butyl lactate,DBl / 161375258.1 28128041-5015 GLB-015PC2024-005 methyl salicylate, propylene carbonate, ethylene carbonate, propylene glycol, ethylene glycol, and combinations thereof.

43. The method of claims 41 or 42, wherein the emulsifier is selected from the group of linear and branched alcohol ethoxylates, castor oil ethoxylates, polysorbate ethoxylates, nonylphenol ethoxylates, tri styryl phenol ethoxylates, coco and tallowamine ethoxylates, tall oil fatty acid ethoxylates, tridecyl alcohol ethoxylates, alkoxylated amides, triblock copolymers, random copolymers, poloxamers, reverse poloxamers, and acrylic copolymers, and combinations thereof.

44. The method claim 38, wherein the pesticidal active ingredient is formulated as a dispersible concentrate (DC).

45. The method of claim 44, wherein the DC comprises the pesticidal active ingredient, a solvent, and an ethoxylated emulsifier.

46. The method of claim 44 or 45, wherein the pesticidal active ingredient comprises about 5% to about 30% by weight of the DC, the solvent comprises about 50% to about 80% by weight of the DC, and the ethoxylated emulsifier comprises about 1% to about 10% by weight of the DC.

47. The method of claim 45 or 46, wherein the solvent is selected from the group of Aromatic 200, d-limonene, acetophenone, ethyl acetate, octanol, butyl lactate, methyl salicylate, propylene carbonate, ethylene carbonate, propylene glycol, ethylene glycol, and combinations thereof.

48. The method of any one of claims 45 to 47, wherein the emulsifier is selected from the group of linear and branched alcohol ethoxylates, castor oil ethoxylates, polysorbate ethoxylates, nonylphenol ethoxylates, tri styrylphenol ethoxylates, coco and tallowamine ethoxylates, tall oil fatty acid ethoxylates, tridecyl alcohol ethoxylates, alkoxylated amides, triblock copolymers, random copolymers, poloxamers, reverse poloxamers, and acrylic copolymers, and combinations thereof.DBl / 161375258.1 29128041-5015 GLB-015PC2024-00549. The method of any one of claims 38 to 48, wherein the EC or DC formulation is prepared by adding the pesticidal active ingredient to a solvent and dissolving at about 50° C and then adding and dissolving an emulsifier.

50. The method of claim 38, wherein the pesticidal active ingredient is formulated as an SC.

51. The method of claim 50, wherein the SC comprises the pesticidal active ingredient, a polymeric dispersing agent, an ethoxylated dispersing agent, an antifoaming agent, a rheology modifier, and a carrier.

52. The method of claim 51, wherein the pesticidal active ingredient comprises about 5% to about 60% by weight of the SC, the polymeric dispersing agent comprises about 1% to about 10% of by weight of the SC, the ethoxylated dispersing agent comprises about 1% to about 10% by weight of the SC, the antifoaming agent comprises about 0.01% to about 0.1% by weight of the SC, the rheology modifier comprises about 0.01% to about 0.1% by weight of the SC, and the carrier comprises about 30% to about 80% by weight of the SC.

53. The method of claim 51 or 52, wherein the antifoaming agent is a trisiloxane antifoaming agent.

54. The method of any one of claims 51 to 53, wherein the rheology modifier is xanthan gum.

55. The method of any one of claims 51 to 54, wherein the carrier is water.

56. The method of any one of claims 51 to 55, wherein the SC is prepared by adding the polymeric dispersing agent and ethoxylated dispersing agent to water and dissolving; adding the antifoaming agent and dissolving; adding the pesticidal active ingredient and mixing under bulking agitation until dispersed; reducing the particle size of the pesticidal active ingredient to an volume average particle size of less than 50 microns; bead milling the resulting formulation to an volume average particle size of about 3 to about 10 microns; and adding the rheology modifier and mixing until fully swelled.DBl / 161375258.1 30128041-5015 GLB-015PC2024-00557. The method of any one of claims 36 to 56, wherein the pesticidal active ingredient comprises Spinosad.

58. The method of any one of claims 36 to 56, wherein the pesticidal active ingredient is selected from the group consisting of ryanodine receptor modulator (IRAC Class 28), an inhibitor of chitin biosynthesis affecting CHS 1 (IRAC Class 15), a nicotinic acetylcholine receptor (nACHR) allosteric modulator-site 1 (IRAC Class 5), an nACHR competitive modulator (IRAC Class 4), a sodium channel modulator (IRAC Class 3), an acetylcholinesterase (ACHE) inhibitor (IRAC Class 1), a GABA-gated chloride channel blocker (IRAC Class 2), a glutamic-gated chloride channel (GLUCL) allosteric modulator (IRAC Class 6), a juvenile hormone receptor modulator (IRAC Class 7), a miscellaneous non-specific (multi-site) inhibitor (IRAC Class 8), a chordotonal organ TRPV channel modulator (IRAC Class 9), a mite growth inhibitor affecting CHS1 (IRAC Class 10), an inhibitor of mitochondrial ATP synthase (IRAC Class 12), an uncoupler of oxidative phosphorylation via disruption of the proton gradient (IRAC Class 13), a nicotinic acetylcholine receptor (nACHR) channel blocker (IRAC Class 14), an inhibitor of chitin biosynthesis type 1 (IRAC Class 16), a molting disruptor dipteran (IRAC Class 17), an ecdysone receptor agonist (IRAC Class 18), an octopamine receptor agonist (IRAC Class 19), a mitochondrial complex III electron transport inhibitor QO site (IRAC Class 20), a mitochondrial complex I electron transport inhibitor (IRAC Class 21), a voltagedependent sodium channel blocker (IRAC Class 22), an inhibitor of acetyl-CoA carboxylase (IRAC Class 23), a mitochondrial complex IV electron transport inhibitor (IRAC Class 24), a mitochondrial transport inhibitor (IRAC Class 25), a chordotonal organ nicotinamidase inhibitor (IRAC Class 29), a GABA-gated chloride channel allosteric modulator (IRAC Class 30), a nicotinic acetylcholine receptor (NACHR) allosteric modulator - site II (IRAC Class 32), a calcium-activated potassium channel (KCa2) modulator (IRAC Class 33), a mitochondrial complex III electron transport inhibitor QI site (IRAC Class 34), or a chordotonal organ modulator - undefined target site (IRAC Class 36), or combinations thereof. In some embodiments, the chemical pesticide is selected from chlorantraniliprole, novaluron, spinosad, acetamiprid, cypermethrin, diflubenzuron, and bifenthrinDBl / 161375258.1 31128041-5015 GLB-015PC 2024-00559. The method of any one of claims 1 to 58, wherein the method and / or system for performing the method is as depicted in Figure 1.

60. A water dispersible granule composition comprising yeast particles and an organic liquid coating, and optionally a pesticidal active ingredient, wherein the yeast particles have a volume average particle size of about 2 to about 10 microns; and wherein the water dispersible granules have granule sizes in the range of about 0.3 to about 10 mm.

61. The water dispersible granule composition of claim 60, having granule sizes of about 4 to about 6 mm; optionally wherein the yeast particles have a volume average particle size of about 2 to about 5 microns.

62. The water dispersible granule composition of claims 60 or 61, further comprising a dispersing agent, a wetting agent, and optionally a filler.

63. The water dispersible granule composition of any one of claims 60 to 62, comprising less than 10% water by weight, or less than about 7.5% water by weight, or about 5% water by weight or less, or about 3% water by weight or less, or about 2% water by weight or less.

64. The water dispersible granule of any one of claims 60 to 63, and produced by the method of any one of claims 1 to 59.DB1 / 161375258.1 32

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