Stable non-aqueous biological pesticides

Abstract

The present invention provides stable, non-aqueous biopesticides. The present invention relates to biopestic formulations of entomopathogenic fungi or their spores. In particular, the present invention discloses oil dispersions containing entomopathogenic fungi or their spores, especially Beauveria bassiana. The present invention also relates to methods for preparing the formulations and their use.

Description

Technical Field

[0001] The present invention relates to biopesticide formulations. In particular, the present invention relates to formulations of entomopathogenic fungi or their spores. The present invention also relates to methods for preparing said formulations and their use.

Background Art

[0002] Pests such as insects, mites, and nematodes are a major problem for agriculture and often significantly reduce productivity. Chemical pesticides are used for pest control, but overuse of chemical pesticides leaves residues in the soil, water, and atmosphere and also has an adverse impact on non-target organisms and the balance of ecosystems. Furthermore, pests can acquire resistance to chemical pesticides, limiting their effectiveness and application range. Concerns among the public about the potential health hazards and increasing costs of chemical pesticides have also led to the search for more environmentally friendly pest management strategies.

[0003] Biopesticides have been developed for use as an alternative to chemical pesticides or, in some cases, as a supplement to chemical pesticides. A biopesticide is an organism (such as a fungus or bacterium) that intervenes in the life cycle of a pest (kills or incapacitates the pest). Biopesticides, which are natural substances, provide a more environmentally friendly solution in pest control and in combination with chemical pesticides. Since some biopesticides (such as fungal spores) are insoluble hydrophobic particles, biopesticides often clog the nozzles of delivery devices or adhere to the inner surface of the delivery tank (often referred to as "staining"). As a solution, it has been proposed to include a surfactant in the formulation of biopesticides, but many surfactants have been found to have a phytotoxic effect on plants, including those important in agriculture. There is an important need for biopesticide formulations that are less phytotoxic to plants, are efficiently delivered, and do not clog the nozzles or adhere to the inner surface of the delivery device during application.

[0004] Biopesticides containing live microorganisms require a very rigorous and precise set of selection criteria, where viability, favorable physiological state, and maintenance of functional biological activity are critical requirements that must be met. As a result of the unique needs of living biological systems, few surfactants, dispersants, and related materials are known that perform satisfactorily, especially in commercial applications. Long-term storage, maintenance of viability, and high toxicity are just some of the properties required for commercially available biological control products, and have not been achieved with known surfactants to date. Specifically, while surfactants generally inhibit spore adhesion and germination, many fungi, such as Metarhizium anisoplie and Beauveria bassiana, metabolize substances such as surfactants, detergents, and wetting agents, so no stable formulation of spores and surfactants is known. In just 72 hours, aqueous suspensions of spores germinate and grow to form mycelial masses in the presence of known surfactants, but this is not useful for commercial insect control. Furthermore, the spores of many entomopathogenic fungi are hydrophobic, making mixing and dispersion in aqueous dilutions difficult. Clearly, little has been done in developing stable and effective aquatic fungal biological control products with a broad activity spectrum and long shelf life. Many of these problems stem from the current inadequacy of formulation technology for biological control agents, especially fungal agents.

[0005] Overall, the lack of biopesticide compositions containing components compatible with viable microorganisms such as entomopathogenic fungi is negatively impacting various fields, including laboratory research (assay development), preparation (processing aids), formulation, and commercial applications. The concentrations of surfactants described in the literature as components of spore suspensions do not exceed 0.5 percent (by volume for liquids, by weight for solids), and reported exposure times are limited to only a few hours. Longer exposures result in loss of viability or the formation of vegetative hyphal morphology through conidial germination and growth.

[0006] Object of the invention While numerous solutions exist for controlling various pests, there is still a need for formulations that not only control pests but also deliver them efficiently to plants without causing phytotoxic effects when applied. For pest control, formulations that can be applied effectively with minimal environmental impact and phytotoxic effects (if any) are desirable (for example, those that do not clog or allow active ingredients and / or other excipients to adhere to surfaces). A stable formulation containing components suitable for entomopathogenic fungi and their spores is now in demand.

[0007] Therefore, the object of the present invention is to provide a material useful for preparing a non-aqueous composition of viable, monodisperse fungal spores that is stable for both long-term and short-term storage and has insecticidal activity.

[0008] A further object of the present invention is to provide spore-compatible components such as oils and surfactants for use in non-aqueous fungal formulations such as oil dispersions and other formulations.

[0009] Another object of the present invention is to provide a material that is stable, harmless to fungi and their spores, does not inhibit germination after application, or alters the viability of fungi and their spores, and is useful for spore preservation.

[0010] Another object of the present invention is to provide materials useful for manipulating and controlling spore survival and proliferation. [Overview of the project]

[0011] In accordance with the purpose of the present invention, one aspect of this disclosure is: a) Entomogenic fungi or their spores, b) Vegetable oil and, c) At least two nonionic surfactants with a hydrophilic-lipophilic balance (HLB) value of 15 or less, The present invention provides a stable, non-aqueous composition containing [the specified element].

[0012] In one embodiment, the stable non-aqueous composition is an oil dispersion.

[0013] In one embodiment, the entomopathogenic fungus or its spores is Beauveria bassiana or its spores.

[0014] In one embodiment, the nonionic surfactant(s) may be a water-soluble nonionic surfactant, a water-insoluble nonionic surfactant, or a combination of a water-soluble nonionic surfactant and a water-insoluble nonionic surfactant. In one embodiment, the nonionic surfactant(s) may be a nonionic polymer surfactant(s).

[0015] In one embodiment, the composition of the present disclosure includes a combination of a nonionic surfactant selected from water-soluble nonionic surfactants and water-insoluble nonionic surfactants and a nonionic polymer surfactant.

[0016] In one embodiment, the nonionic surfactant has a hydrophilic-lipophilic balance (HLB) value of 12 or less. In a preferred embodiment, the nonionic surfactant has a hydrophilic-lipophilic balance (HLB) value of 4 to 12. In another embodiment, one of the two nonionic surfactants has an HLB value in the range of 8 to 12, and the other nonionic surfactant has an HLB value in the range of 4 to 8.

[0017] In one embodiment, the weight ratio of the two nonionic surfactants in the composition is in the range of 1:100 to 100:1.

[0018] One aspect of this disclosure is, a) an amount of entomopathogenic fungi or their spores in the range of approximately 0.1% w / w to approximately 30% w / w of the total weight of the composition, b) A vegetable oil in an amount ranging from approximately 10% w / w to approximately 99.8% w / w of the total weight of the composition, c) At least two nonionic surfactants with a hydrophilic-lipophilic balance (HLB) value of 15 or less, in an amount ranging from approximately 0.1% w / w to approximately 60% w / w of the total weight of the composition, The present invention provides a stable, non-aqueous composition containing [the specified element].

[0019] One aspect of the present disclosure provides a method for preparing a stable non-aqueous composition, the method comprising: i) mixing a vegetable oil and at least two nonionic surfactants having a hydrophilic-lipophilic balance (HLB) value of 15 or less under high shear; ii) adding an entomopathogenic fungus or its spores to the mixture obtained in step i); iii) homogenizing the mixture of step ii) to obtain a stable composition. It includes.

[0020] In one embodiment, the method optionally includes adding a rheology modifier or an anionic surfactant to the mixture obtained in step i) under high shear before adding the entomopathogenic fungus or its spores.

[0021] One aspect of the present disclosure is i) mixing a vegetable oil and at least two nonionic surfactants having a hydrophilic-lipophilic balance (HLB) value of 15 or less under high shear; ii) adding an entomopathogenic fungus or its spores to the mixture obtained in step i); iii) homogenizing the mixture of step ii) to obtain a stable composition. It provides an oil dispersion obtained by a method including.

[0022] One aspect of the present disclosure is a) an entomopathogenic fungus or its spores; b) a vegetable oil; c) at least two nonionic surfactants having a hydrophilic-lipophilic balance (HLB) value of 15 or less. It provides a method for improving the storage stability and viability of an entomopathogenic fungus or its spores, which includes preparing a non-aqueous composition containing.

[0023] One aspect of the present disclosure provides a method for controlling plant pests, the method comprising applying one or more plant pests with a) an entomopathogenic fungus or its spores; b) Vegetable oil and, c) At least two nonionic surfactants with a hydrophilic-lipophilic balance (HLB) value of 15 or less, The process includes the step of bringing the mixture into contact with a composition containing the following: [Modes for carrying out the invention]

[0024] As used herein, the singular forms "one," "one," and "the" also include the plural form unless the context clearly indicates otherwise. As used herein, the terms "active," "active ingredient," and "agricultural active ingredient" mean any organism or chemical element, molecule, compound, or mixture thereof that has biological activity in seeds, plants, or diseases or pests of seeds or plants. Such active ingredients include, but are not limited to, insecticides, herbicides, fertilizers, plant growth regulators, drugs, dyes, bioattractants, fragrances, and pheromones.

[0025] The biopesticides (i.e., compositions) used in the embodiments disclosed herein comprise an agriculturally suitable carrier, an effective amount of at least one fungal pesticide (e.g., two or more, e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.), and at least one surfactant (e.g., two or more, e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.). In preferred embodiments, the at least one surfactant is selected from sorbitan fatty acid esters, sorbitol ethoxylate esters, alcohol ethoxylates, and combinations thereof.

[0026] In this specification, the term “fungal insecticide” means a fungal organism in a vegetative or dormant state (e.g., spore) that is pathogenic to target pests such as insects, mites, or nematodes. The term “spore” as used herein has the common meaning known and understood by those skilled in the art and refers to a protected microorganism in a dormant state.

[0027] As used herein, the term “entomophagetic” means that a fungal insecticide is pathogenic to at least one target insect species. As used herein, “entomophagetic fungus” is a fungus that has the ability to attack, infect, kill, incapacitate, disease, and / or injure insects and can therefore be used to control insect infestations by adversely affecting the viability or growth of target insects.

[0028] As used herein, “exogenously applied” means that the cuticle-degrading enzyme is applied independently of (i.e., as a separate component from) the enzymes produced by the compositions and fungicides disclosed herein.

[0029] The term "pest" refers to animals of the scientific classification (phylum) arthropods, which include Insecta (such as whiteflies, thrips, and weevils) and Arachnida (including, but not limited to, mites, ticks, spiders, and other invertebrates).

[0030] As used herein, the terms “control” or “to control” mean, for example, “control” of a pest or pest population, or “control” of a pest or pest population, or as used in the following phrases, “to control” a pest means the prevention, reduction, eradication, suppression of proliferation, or removal of a pest or pest population as defined herein. In fact, as used herein, “control” or “to control” means signs of success in the prevention, eradication, suppression, removal, reduction, or improvement of a pest or pest population.

[0031] As used herein, the terms “effective dose,” “effective concentration,” or “effective dosage” are defined as the amount, concentration, or dosage of a fungal insecticide sufficient to cause infection in a pest and subsequently lead to control of the pest. The absolute value of the actual effective dosage will vary depending on, but will not be limited to, factors such as the mortality rate of the target pest relative to the amount of fungal insecticide applied, synergistic or antagonistic effects between other active or inactive components that may increase or decrease the activity of the fungal insecticide, the life stage and species-specific susceptibility of the pest, and the stability of the fungal insecticide in the composition. The “effective dose,” “effective concentration,” or “effective dosage” of a fungal insecticide may be determined, for example, by a standard dose-response experiment. As used herein, the term “agriculturally beneficial component” means any agent or combination of agents that can produce or provide beneficial and / or useful effects in agriculture. As used herein, terms such as “agriculturally beneficial microorganisms,” “agriculturally beneficial bacteria,” and “agriculturally beneficial bacteria” are intended to mean any microorganism (e.g., bacteria, fungi, etc., or combination thereof) that is capable of causing or providing beneficial and / or useful effects in agriculture (e.g., promoting plant growth, providing fungicidal activity, etc.), whether the microorganism is in a vegetative state or in spore form.

[0032] As used herein, terms such as “spore” and “microbial spore” have their ordinary meanings as are well known and understood by those skilled in the art. As used herein, “spore” and “microbial spore” refer to a protected microorganism in a dormant state.

[0033] As used herein, the term "inoculum" means any form of microbial cells or spores that can grow on or within soil when conditions such as temperature and humidity are favorable for microbial growth.

[0034] As used herein, the terms “plants” and “parts of plants” are intended to refer to all plants and plant populations, including desirable wild plants and undesirable wild plants or crop plants (including naturally occurring crop plants). Crop plants are plants obtained by conventional plant breeding and optimization methods, biotechnology and genetic engineering methods, or combinations thereof, and include genetically modified plants and plant cultivars, whether protected or not by plant breeders’ rights. Parts of plants are understood to mean all parts and organs of plants above and below ground, such as buds, leaves, flowers, and roots, but examples may include leaves, needles, stems, trunks, flowers, fruiting bodies, fruits, seeds, roots, tubers, and rhizomes. Parts of plants also include harvested products, vegetative and reproductive products (e.g., cuttings, tubers, rhizomes, lateral buds, seeds, etc.).

[0035] As used herein, the term “leaf” means all parts and organs of the above-ground parts of a plant. Examples, not limited to, include leaves, needles, stems, trunks, flowers, fruiting bodies, and fruits. As used herein, the terms “foliar application,” “applied to the foliar surface,” and variations thereof are intended to include the application of an active ingredient to the leaves or above-ground parts of a plant (e.g., the leaves of a plant). Application may be carried out by any means known in the art (e.g., spraying of the active ingredient).

[0036] As used herein, the term “source” of a particular element means a compound of that element that, at least under the soil conditions under consideration, is not fully available for absorption by plants.

[0037] As used herein, the term “nutrient” means any nutrient necessary for the growth, health, and / or development of a plant (e.g., vitamins, macrominerals, micronutrients, trace minerals, organic acids, etc.).

[0038] As used herein, the term “herbicide” means any chemical or combination of chemicals capable of killing weeds and / or inhibiting their growth (the inhibition being reversible under certain conditions).

[0039] As used herein, the term “fungicide” refers to any agent or combination of agents that can kill and / or inhibit the growth of fungi.

[0040] As used herein, the term “insecticides” means any agent or combination of agents that can kill one or more species of insects and / or inhibit the reproduction of one or more species of insects.

[0041] As used herein, the term “nematicidal agent” means any agent or combination of agents that can kill one or more species of nematodes and / or inhibit the growth of one or more species of nematodes.

[0042] As used herein, the term “acaricide” refers to any agent or combination of agents that can kill one or more species of mites and / or inhibit the proliferation of one or more species of mites. As used herein, the term “biostimulant” refers to any agent or combination of agents that can enhance metabolic or physiological processes in plants and soil.

[0043] As used throughout this specification, the terms “parts by weight” and “percent by weight” are used interchangeably herein, and the respective percentage by weight of individual components is expressed as a percentage by weight based on the total weight of the particular composition in which it forms part.

[0044] The inventors have discovered that fungal spore formulations, which are often suspended in oil, do not disperse well when diluted with water. While not bound by theory, it is thought that if the oil phase is not dispersed sufficiently, the size of the oil droplets will increase, and the hydrophobic fungal spores will be attracted to the oil droplets and / or become completely contained within them.

[0045] These droplets are attracted to the inner walls / surfaces of sprayer tanks (especially plastic containers commonly used in agriculture), resulting in insufficient and inefficient dispersion of active ingredients (such as fungal spores), clogging of spraying equipment (nozzles, hoses, etc.), and difficulty in cleaning tanks and other spraying and delivery devices. Various surfactant systems have been applied to address this problem, but plant toxicity remains a concern. Surprisingly and unexpectedly, the inventors discovered that appropriately combining surfactants in specific proportions (often in very small amounts compared to others) reduces the overall plant toxicity of a particular formulation while simultaneously overcoming the known challenges encountered in properly delivering the active ingredients when diluted with water and applied.

[0046] Furthermore, by modifying the carrier to increase the dispersion and / or suspension of biopesticides in aqueous solutions, it is believed that biopesticides can be efficiently delivered (e.g., through a sprayer) without the active substance adhering to / fixing to the inner wall of the delivery device or clogging delivery components (e.g., sprayer nozzle, sprayer tube, etc.).

[0047] The surfactants suitable for biopesticides disclosed herein minimize any adverse effects on the viability of fungal pesticides.

[0048] Accordingly, this specification discloses compositions that act as biopesticides and methods that provide an improved practical approach to controlling damage to crops caused by pest populations. The biopesticides described herein have the advantage of controlling pests, but also the additional advantage that when fully formulated they are delivered efficiently (i.e., the active substance and / or other excipients are not retained inside the holding tank, for example, reducing the adhesion / sticking of the active substance and / or other excipients to the inner surface of the tank, and preventing clogging of the delivery device or its components (nozzle or hose)), and any phytotoxic damage to crops treated with the fully formulated biopesticides is minimal, if any.

[0049] In one embodiment, this disclosure, a) Entomogenic fungi or their spores, b) Vegetable oil and, c) At least two nonionic surfactants with a hydrophilic-lipophilic balance (HLB) value of 15 or less, The present invention provides a stable non-aqueous composition containing [a certain element].

[0050] In a preferred embodiment, the stable non-aqueous composition is an oil dispersion.

[0051] In one embodiment, the composition is a fungal insecticide composition. In one embodiment, the spores of an entomopathogenic fungus act as a fungal insecticide. In one embodiment, the composition is a biological insecticide composition containing a fungal insecticide.

[0052] In one embodiment, the spores of entomopathogenic fungi belong to the phyla Ascomycota, Alternaria, Beauveria, Lecanicillium, Metallidium, Verticillium, Trichoderma, Aspergillus, Nomuraea, Paesilomyces, Isalia, Hirsterella, Fusarium, Cordyceps, Entomovhosola, Zoufsola, Pandora, Entomophaga, Entomovsolareth, Zygomycota, and combinations thereof.

[0053] In one embodiment, non-limiting examples of entomopathogenic fungi that can be used in the compositions disclosed herein include: Coelomisidia, Myophagus, Coeremomyces, Lagenidium, Leptolignia, Coutia, Sporodiniella, Conidiobolus, Entomophaga, Entomofsora, Ellinia, Massospora, Melistacrum, Neodigites, Pandora, Zoufsora, Blastodendrion, Metchnikowia, Mycoderma, Ascophaera, Cordyceps, Torviella, Nectaria, Hypocrinella, Caronectaria, Filariomyces, Hesperomyces , Trenomyces, Myriangium, Podnectria, Acanthomyces, Ashersonia, Aspergillus, Beauveria, Chrysinomyces, Engyodontium, Fusarium, Giberla, Hirsterella, Hymenostylbe, Isaria, Metallidium, Nomuraea, Paesilomyces, Paraizaria, Pleurodesmospora, Polycephalomyces, Pseudogiberra, Sorosporella, Stilbella, Tetranacrium, Thyraclidium, Tripocladium, Verticillium, Agerita, Phylobasidiera, Septobasidium, Uredinella, and combinations thereof.

[0054] In one embodiment, non-limiting examples of entomopathogenic fungal species that may be useful as fungal insecticides in the biopesticides described herein include Trichoderma hamatum, Trichoderma hazardum, Alterna cassier, Fusarium lateritum, Fusarium solani, Lecanicillium lecanii, Aspergillus paracithus, Verticillium lecanii, Metahydium anisoplie, and Beauveria bassiana. In one embodiment, the compositions disclosed herein may comprise any of the above fungal insecticides and any combination thereof. In another embodiment, the fungal insecticide is stable and maintains a sufficient effective amount of activity at the time of use. In one embodiment, the fungal insecticide is present in the composition in the form of stable spores.

[0055] In yet another embodiment, the entomopathogenic fungus comprises at least one fungal insecticide derived from the genus Beauveria, such as Beauveria bassiana. In at least one embodiment, the entomopathogenic fungus comprises strain Beauveria bassiana. In another embodiment, the entomopathogenic fungus further comprises spores of strain Beauveria bassiana. Those skilled in the art will understand that the compositions of the present invention may comprise any strain of Beauveria bassiana, or spores of any strain of Beauveria bassiana, and that the present invention is not particularly limited to the strain used.

[0056] In a preferred embodiment, the entomopathogenic fungus comprises at least one fungal insecticide containing Beauveria bassiana TK. In another embodiment, the entomopathogenic fungus comprises at least one fungal insecticide containing spores of Beauveria bassiana TK. In a preferred embodiment, the entomopathogenic fungus comprises at least one fungal insecticide containing Beauveria bassiana strain ATCC-74040. In another embodiment, the entomopathogenic fungus comprises at least one fungal insecticide containing spores of Beauveria bassiana strain ATCC-74040. In another embodiment, the entomopathogenic fungus comprises at least one fungal insecticide containing Beauveria bassiana strain ATCC-74250. In another embodiment, the entomopathogenic fungus comprises at least one fungal insecticide containing Beauveria bassiana strain CNCM I-867. In another embodiment, the entomopathogenic fungus comprises at least one fungal insecticide containing Beauveria bassiana strain CNCM I-2960. In yet another embodiment, the entomopathogenic fungus comprises at least one fungal insecticide containing spores of Beauveria bassiana strain ATCC-74250. In yet another embodiment, the entomopathogenic fungus comprises at least one fungal insecticide containing spores of Beauveria bassiana strain CNCM I-867. In yet another embodiment, the entomopathogenic fungus comprises at least one fungal insecticide containing spores of Beauveria bassiana strain CNCM I-2960. In yet another embodiment, the entomopathogenic fungus comprises at least one fungal insecticide containing a mixture of Beauveria bassiana strain ATCC-74040 and Beauveria bassiana strain ATCC-74250. In yet another embodiment, the entomopathogenic fungus comprises at least one fungal insecticide containing a mixture of spores of Beauveria bassiana strain ATCC-74040 and Beauveria bassiana strain ATCC-74250. In yet another embodiment, the entomopathogenic fungus comprises at least one fungal insecticide containing a mixture of spores of Beauveria bassiana strain CNCM I-867 and Beauveria bassiana strain CNCM I-2960. In yet another embodiment, the entomopathogenic fungus comprises at least one fungal insecticide containing a mixture of spores of Beauveria bassiana strain CNCM I-867 and Beauveria bassiana strain CNCM I-2960.In another embodiment, the entomopathogenic fungus described herein may comprise a combination of various strains of Beauveria bassiana, or spores of various strains of Beauveria bassiana. In another embodiment, the entomopathogenic fungus described herein may comprise a combination of fungi. In one embodiment, the entomopathogenic fungus may comprise two or more fungal insecticides that are different strains of the same species. In another embodiment, the entomopathogenic fungus may comprise at least two different fungal insecticides that are strains of different species.

[0057] In one embodiment, the vegetable oil is selected from olive oil, kapok oil, castor oil, palm oil, sunflower oil, camellia oil, coconut oil, sesame oil, corn oil, rice bran oil, peanut oil, cottonseed oil, soybean oil, rapeseed oil, linseed oil, tung oil, canola oil, corn oil, or a combination thereof. In a preferred embodiment, the vegetable oil is sunflower oil. In a preferred embodiment, the vegetable oil is corn oil. In a preferred embodiment, the vegetable oil is soybean oil.

[0058] The compositions described herein may comprise at least two nonionic surfactants having a hydrophilic-lipophilic balance (HLB) value of 15 or less. In one embodiment, the nonionic surfactant has an HLB value of 12 or less. In a preferred embodiment, the nonionic surfactant has an HLB value of 4 to 12. In another embodiment, one of the two nonionic surfactants has an HLB value in the range of 8 to 12, and the other nonionic surfactant has an HLB value in the range of 4 to 8.

[0059] In one embodiment, the nonionic surfactant is selected from a water-soluble nonionic surfactant, a water-insoluble nonionic surfactant, a nonionic polymer surfactant, or a combination thereof.

[0060] In one embodiment, the nonionic surfactant may be a water-soluble nonionic surfactant, a water-insoluble nonionic surfactant, or a combination of a water-soluble nonionic surfactant and a water-insoluble nonionic surfactant. In one embodiment, the HLB value of the water-soluble nonionic surfactant and the water-insoluble nonionic surfactant is 15 or less, preferably 12 or less. In one embodiment, the HLB value of the water-soluble nonionic surfactant and the water-insoluble nonionic surfactant is in the range of 4 to 12. In another embodiment, the preferred HLB value of the water-insoluble nonionic surfactant is in the range of 8 to 12. In yet another embodiment, the preferred HLB value of the water-insoluble nonionic surfactant is 10.

[0061] In one embodiment, the nonionic surfactant(s) may be a nonionic polymer surfactant(s). In one embodiment, the HLB value of the nonionic polymer surfactant is 15 or less, preferably 12 or less. In another embodiment, the HLB value of the nonionic polymer surfactant is in the range of 4 to 12. In yet another embodiment, the preferred HLB value of the nonionic polymer surfactant is in the range of 4 to 8. In yet another embodiment, the preferred HLB value of the nonionic polymer surfactant is 6.

[0062] In one embodiment, the composition of the present disclosure comprises at least two nonionic surfactants. In another embodiment, the composition comprises at least two water-insoluble nonionic surfactants. In yet another embodiment, the composition comprises at least two water-soluble nonionic surfactants. In yet another embodiment, the composition comprises at least two nonionic polymeric surfactants.

[0063] In one embodiment, the composition comprises one or more water-insoluble nonionic surfactants. In yet another embodiment, the composition comprises one or more water-soluble nonionic surfactants. In yet another embodiment, the composition comprises one or more nonionic polymer surfactants. In yet another embodiment, the composition comprises one or more water-insoluble nonionic surfactants and one or more water-soluble nonionic surfactants. In yet another embodiment, the composition comprises one or more water-insoluble nonionic surfactants and one or more nonionic polymer surfactants. In yet another embodiment, the composition comprises one or more water-soluble nonionic surfactants and one or more nonionic polymer surfactants.

[0064] The following table shows the surfactants in the composition. At least two nonionic surfactants are indicated as Surfactant 1 and Surfactant 2, and their respective HLB values ​​are shown.

[0065] [Table A]

[0066] In one embodiment, the composition of the present disclosure includes a combination of a nonionic surfactant selected from water-soluble nonionic surfactants and water-insoluble nonionic surfactants, and a nonionic polymer surfactant. In one embodiment, the HLB value of the nonionic surfactant is in the range of 4 to 12. In a preferred embodiment, the HLB value of the water-insoluble nonionic surfactant is in the range of 8 to 12. In a preferred embodiment, the preferred HLB value of the nonionic polymer surfactant is 6.

[0067] In one embodiment, the composition of the present disclosure includes a combination of a water-insoluble nonionic surfactant and a nonionic polymeric surfactant.

[0068] In one embodiment, the nonionic surfactant(s) of the composition is a combination of a water-soluble nonionic surfactant and a nonionic polymer surfactant.

[0069] In preferred embodiments, the nonionic polymer surfactant is selected from polyalkylene oxide block copolymers, butyl block copolymers, nonionic block copolymers, acrylic copolymer solutions, nonionic random polymer polymers, polyoxyethylene polyarylphenols, nonionic polymer dispersants, polyisobutylene succinic anhydride-polyethylene glycol, and combinations thereof. In more preferred embodiments, the composition comprises one or more nonionic polymeric surfactants selected from polyisobutylene succinic anhydride-polyethylene glycol, polyalkylene oxide block copolymer, polyalkylene glycol ether, polyoxyalkylene glycol aliphatic alcohol ether, polyoxyethylene alkyl ether, block copolymer of polyethylene glycol and 12-hydroxystearic acid, polymethyl methacrylate-polyethylene glycol graft copolymer, succinic anhydride-polyethylene glycol, polyoxyethylene polyarylphenol, block copolymer of poly(12-hydroxystearic acid) and poly(ethylene oxide), block copolymer of poly(12-hydroxystearic acid) and poly(ethylene oxide) having a molecular weight of 1500 to 10000 (also known as polyethylene glycol), alkylbenzene sulfonates, alkanoyl taurates, and alkanoyl sarcosinates, and combinations thereof.

[0070] In one embodiment, the water-insoluble nonionic surfactant is selected from alkyl and aryl:glycerol ethers, glycol ethers, ethanolamides, sulfonylamides, alcohols, amides, alcohol ethoxylates, ethoxylated oils such as castor oil ethoxylate, glycerol esters, glycol esters, ethoxylates of glycerol esters and glycol esters, sugar-based alkyl polyglycosides, polyoxyethylene-converted fatty acids, alkanolamine condensates, alkanolamides, tertiary acetylene glycols, polyoxyethylene-converted mercaptans, carboxylic acid esters, polyoxyethylene-converted polyoxypropylene glycols, sorbitan fatty acid esters, sorbitol ethoxylate esters, EO / PO block copolymers (EO is ethylene oxide, PO is propylene oxide), EO polymers and copolymers, polyamines, polyvinylpyrrolidone, sulfosuccinate esters such as sodium dioctyl sulfosuccinate, and combinations thereof.

[0071] Non-limiting examples of commercially available water-insoluble nonionic surfactants suitable for the compositions described herein include polyoxyethylene (30) oleate laurate, polyoxyethylene (40) sorbitol hexaoleate, polyoxyethylene (50) sorbitol hexaoleate, alcohol ethoxylates, sorbitan fatty acid esters, polyoxyethylene (40) sorbitol oleate, and combinations thereof.

[0072] In one embodiment, the composition of the Disclosure comprises at least one water-insoluble nonionic surfactant. In another embodiment, the composition of the Disclosure comprises at least one water-insoluble nonionic surfactant selected from sorbitan fatty acid esters, sorbitol ethoxylate esters, and combinations thereof. Non-limiting examples of sorbitan fatty acid esters suitable for the composition of the Disclosure include sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, sorbitan tristearate, sorbitan monooleate, sorbitan trioleate, and combinations thereof. Non-limiting examples of sorbitol ethoxylate esters suitable for the composition of the Disclosure include polyoxyethylene (40) sorbitol oleate, polyoxyethylene (40) sorbitol hexaoleate, polyoxyethylene (50) sorbitol hexaoleate, polyoxyethylene (30) oleate laurate, polyoxyethylene sorbitan monolaurate, and combinations thereof.

[0073] In another preferred embodiment, the composition comprises one or more sorbitan fatty acid esters selected from sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, sorbitan tristearate, sorbitan monooleate, sorbitan trioleate, and combinations thereof. In yet another preferred embodiment, the composition comprises one or more sorbitol ethoxylate esters selected from polyoxyethylene (40) sorbitol oleate, polyoxyethylene (40) sorbitol hexaoleate, polyoxyethylene (50) sorbitol hexaoleate, polyoxyethylene (30) oleate laurate, and combinations thereof.

[0074] In yet another preferred embodiment, the composition comprises at least one sorbitan fatty acid ester selected from sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, sorbitan tristearate, sorbitan monooleate, sorbitan trioleate, and combinations thereof, as well as a sorbitol ethoxylate ester, the sorbitol ethoxylate ester being selected from polyoxyethylene (40) sorbitol oleate, polyoxyethylene (40) sorbitol hexaoleate, polyoxyethylene (50) sorbitol hexaoleate, polyoxyethylene (30) oleate laurate, and combinations thereof.

[0075] In another embodiment, the composition comprises sorbitan monostearate. In yet another embodiment, the composition comprises sorbitan monooleate. In yet another embodiment, the composition comprises polyoxyethylene sorbitol hexaoleate. In yet another embodiment, the composition comprises polyoxyethylene (40) sorbitol hexaoleate. In a preferred embodiment, the composition comprises sorbitan monostearate, sorbitan monooleate, polyoxyethylene (40) sorbitol hexaoleate, and combinations thereof. In another preferred embodiment, the composition comprises sorbitan monostearate, sorbitan monooleate, and combinations thereof. In yet another preferred embodiment, the composition comprises sorbitan monostearate, polyoxyethylene (40) sorbitol hexaoleate, and combinations thereof. In yet another preferred embodiment, the composition comprises sorbitan monooleate, polyoxyethylene (40) sorbitol hexaoleate, and combinations thereof.

[0076] In one embodiment, the water-soluble nonionic surfactant is selected from sorbitan fatty acid alcohol ethoxylates, alkoxylated diethylethanolamines, sorbitan fatty acid ester ethoxylates, and combinations thereof. In one embodiment, the composition comprises at least one water-soluble nonionic surfactant which is a linear primary, secondary, or branched alcohol ethoxylate having the following formula: RO(CH2CH20)nH (where R is the hydrocarbon chain length and n is the average number of moles of ethylene oxide). In one embodiment, R may be a linear primary, secondary, or branched alcohol ethoxylate having a hydrocarbon chain length in the range of C9 to C16 and n in the range of 6 to 13. In another embodiment, the composition comprises at least one alcohol ethoxylate where R is a linear C9-C11 hydrocarbon chain length and n is 6. In yet another embodiment, if the composition of the present disclosure comprises two or more water-soluble surfactants, the water-soluble surfactants have substantially the same carbon chain length.

[0077] In one embodiment, commercially available water-soluble nonionic surfactants suitable for the compositions of the present disclosure include alcohol ethoxylates, polyoxyethylene sorbitol esters, polyoxyethylene (4) sorbitan monolaurate, polyoxyethylene sorbitan monopalmitate, polyoxyethylene (20) sorbitan monostearate, polyethylene sorbitol esters, polyoxyethylene (20) sorbitan monooleate, 22-mol ethoxylates of linear primary 12-14 carbon alcohols, and combinations thereof.

[0078] In one embodiment, the composition of the present disclosure comprises at least one water-soluble nonionic surfactant selected from the group consisting of alcohol ethoxylates, polyoxyethylene sorbitol esters, polyoxyethylene (4) sorbitan monolaurate, polyoxyethylene sorbitan monopalmitate, polyoxyethylene (20) sorbitan monostearate, polyethylene sorbitol esters, polyoxyethylene (20) sorbitan monooleate, 22-mol ethoxylates of linear primary 12-14 carbon alcohols, and combinations thereof.

[0079] In preferred embodiments, the compositions of the present disclosure include at least one sorbitan fatty acid ester ethoxylate selected from the group consisting of alcohol ethoxylates, polyoxyethylene sorbitol esters, polyoxyethylene (4) sorbitan monolaurate, polyoxyethylene sorbitan monopalmitate, polyoxyethylene (20) sorbitan monostearate, polyethylene sorbitol esters, polyoxyethylene (20) sorbitan monooleate, 22-mol ethoxylates of linear primary 12-14 carbon-number alcohols, and combinations thereof.

[0080] In yet another embodiment, the compositions of the present disclosure comprise at least one alcohol ethoxylate, at least one sorbitan fatty acid ester ethoxylate, or a combination thereof. In yet another embodiment, the compositions of the present disclosure comprise at least one water-soluble nonionic surfactant selected from the group consisting of alcohol ethoxylates, polyoxyethylene sorbitol esters, polyoxyethylene (4) sorbitan monolaurate, polyoxyethylene sorbitan monopalmitate, polyoxyethylene (20) sorbitan monostearate, polyethylene sorbitol esters, polyoxyethylene (20) sorbitan monooleate, 22-mol ethoxylates of linear primary 12-14 carbon alcohols, and combinations thereof.

[0081] In one embodiment, the weight ratio of the two nonionic surfactants in the composition is in the range of 1:100 to 100:1. In one embodiment, the weight ratio of the two nonionic surfactants in the composition is in the range of 1:50 to 50:1. In one embodiment, the weight ratio of the two nonionic surfactants in the composition is in the range of 1:20 to 20:1. In one embodiment, the weight ratio of the two nonionic surfactants in the composition is in the range of 1:10 to 10:1. In one embodiment, the weight ratio of the two nonionic surfactants in the composition is in the range of 1:7.5 to 7.5:1. In one embodiment, the weight ratio of the two nonionic surfactants in the composition is in the range of 1:5 to 5:1. In one embodiment, the weight ratio of the two nonionic surfactants in the composition is in the range of 1:3 to 3:1.

[0082] In one embodiment, the composition of the present disclosure comprises one or more nonionic surfactants selected from water-soluble nonionic surfactants and water-insoluble nonionic surfactants, and one or more nonionic polymeric surfactants, wherein the HLB values ​​of the nonionic surfactants are in the range of 4 to 12.

[0083] In one embodiment, the entomopathogenic fungus or its spores constitute about 0.1% w / w to about 30% w / w of the total weight of the composition. In a preferred embodiment, the entomopathogenic fungus or its spores constitute about 0.5% w / w to about 20% w / w of the total weight of the composition. In yet another embodiment, the entomopathogenic fungus or its spores constitute about 0.7% w / w to about 10% w / w of the total weight of the composition. In yet another embodiment, the entomopathogenic fungus or its spores constitute about 1% w / w to about 5% w / w of the total weight of the composition.

[0084] In one embodiment, Beauveria bassiana or its spores constitute about 0.1% w / w to about 30% w / w of the total weight of the composition. In a preferred embodiment, Beauveria bassiana or its spores constitute about 0.5% w / w to about 20% w / w of the total weight of the composition. In yet another embodiment, Beauveria bassiana or its spores constitute about 0.7% w / w to about 10% w / w of the total weight of the composition. In yet another embodiment, Beauveria bassiana or its spores constitute about 1% w / w to about 5% w / w of the total weight of the composition. In one embodiment, the composition is about 1 × 10 7 CFU ~ approximately 1 x 10 11 The CFU Beauveria bassiana contains Beauveria bassiana spores. In a preferred embodiment, the composition is about 1 × 10 9 Contains CFU Beauveria bassiana spores.

[0085] In one embodiment, the vegetable oil constitutes about 10% w / w to about 99.8% w / w of the total weight of the composition. In a preferred embodiment, the vegetable oil constitutes about 20% w / w to about 99% w / w of the total weight of the composition. In a preferred embodiment, the vegetable oil constitutes about 30% w / w to about 90% w / w of the total weight of the composition. In a preferred embodiment, the vegetable oil constitutes about 40% w / w to about 85% w / w of the total weight of the composition. In a preferred embodiment, the vegetable oil constitutes about 50% w / w to about 80% w / w of the total weight of the composition. In a preferred embodiment, the vegetable oil constitutes about 60% w / w to about 85% w / w of the total weight of the composition. In a preferred embodiment, the vegetable oil constitutes about 60% w / w to about 75% w / w of the total weight of the composition.

[0086] In one embodiment, sunflower oil constitutes about 10% w / w to about 99.8% w / w of the total weight of the composition. In a preferred embodiment, sunflower oil constitutes about 20% w / w to about 99% w / w of the total weight of the composition. In a preferred embodiment, sunflower oil constitutes about 30% w / w to about 90% w / w of the total weight of the composition. In a preferred embodiment, sunflower oil constitutes about 40% w / w to about 85% w / w of the total weight of the composition. In a preferred embodiment, sunflower oil constitutes about 50% w / w to about 80% w / w of the total weight of the composition. In a preferred embodiment, sunflower oil constitutes about 60% w / w to about 85% w / w of the total weight of the composition. In a preferred embodiment, sunflower oil constitutes about 60% w / w to about 75% w / w of the total weight of the composition.

[0087] In one embodiment, the nonionic surfactant constitutes about 0.1% w / w to about 60% w / w of the total weight of the composition. In a preferred embodiment, the nonionic surfactant constitutes about 0.5% w / w to about 50% w / w of the total weight of the composition. In yet another embodiment, the nonionic surfactant constitutes about 1% w / w to about 40% w / w of the total weight of the composition. In yet another embodiment, the nonionic surfactant constitutes about 1.5% w / w to about 30% w / w of the total weight of the composition.

[0088] In one embodiment, the water-insoluble nonionic surfactant constitutes about 0.1% w / w to about 60% w / w of the total weight of the composition. In a preferred embodiment, the water-insoluble nonionic surfactant constitutes about 0.5% w / w to about 50% w / w of the total weight of the composition. In yet another embodiment, the water-insoluble nonionic surfactant constitutes about 1% w / w to about 40% w / w of the total weight of the composition. In yet another embodiment, the water-insoluble nonionic surfactant constitutes about 2% w / w to about 30% w / w of the total weight of the composition. In yet another embodiment, the water-insoluble nonionic surfactant constitutes about 5% w / w to about 25% w / w of the total weight of the composition.

[0089] In one embodiment, the nonionic polymer surfactant constitutes about 0.1% w / w to about 60% w / w of the total weight of the composition. In a preferred embodiment, the nonionic polymer surfactant constitutes about 0.5% w / w to about 50% w / w of the total weight of the composition. In yet another embodiment, the nonionic polymer surfactant constitutes about 1% w / w to about 40% w / w of the total weight of the composition. In yet another embodiment, the nonionic polymer surfactant constitutes about 1% w / w to about 30% w / w of the total weight of the composition. In yet another embodiment, the nonionic polymer surfactant constitutes about 1% w / w to about 20% w / w of the total weight of the composition. In yet another embodiment, the nonionic polymer surfactant constitutes about 1% w / w to about 10% w / w of the total weight of the composition. In yet another embodiment, the nonionic polymer surfactant constitutes about 1% w / w to about 5% w / w of the total weight of the composition.

[0090] The compositions of this disclosure may further include one or more arbitrary components that are physically and / or chemically compatible with the components described herein. Examples of arbitrary components, not limited to these, include anti-settling agents (rheological additives, also referred to herein as rheological modifiers), agriculturally beneficial components (e.g., enzymes, beneficial plant signaling molecules, beneficial microorganisms, insecticides, fungicides, nematicides, nutrients, etc.), insect growth regulators, electrostatic carriers, preservatives, fillers, pH adjusters, stabilizers, builders, buffers, antioxidants, water absorbers, foaming agents, humectants, wetting agents, UV protectants, solvents, nutritional additives, and combinations thereof. Such components are known to those skilled in the art.

[0091] In at least one embodiment, the compositions of the Disclosure may optionally comprise one or more anti-settling agents / rheological modifiers. Alternatively, one or more anti-settling agents may be applied simultaneously with or sequentially to the biopesticides disclosed herein. The one or more anti-settling agents may comprise any agents capable of keeping insoluble particles (i.e., fungal pesticide spores) uniformly suspended in a liquid solution (i.e., preventing the sedimentation of the insoluble particles).

[0092] Non-limiting examples of anti-settling agents suitable for the compositions of this disclosure include polyvinyl acetate, polyvinyl alcohols of different degrees of hydrolysis, polyvinylpyrrolidone, polyacrylates, water-soluble or dispersible acrylates, polyol or polyester-based paint binders, copolymers of two or more monomers such as acrylic acid, methacrylic acid, itaconic acid, maleic acid, fumaric acid, maleic anhydride, and vinylpyrrolidone, ethylenically unsaturated monomers such as ethylene, butadiene, isoprene, chloroprene, styrene, divinylbenzene, α-methylstyrene, or β-methylstyrene, further vinyl halides such as vinyl chloride and vinylidene chloride, vinyl esters such as vinyl acetate, vinyl propionate, or vinyl stearate, and further esters of vinyl methyl ketone or acrylic acid or methacrylic acid with monohydric alcohols. Alternatively, examples include polyols such as methyl acrylate, methyl methacrylate, ethyl acrylate, ethylene methacrylate, lauryl acrylate, lauryl methacrylate, decyl acrylate, N,N-dimethylaminoethyl methacrylate, 2-hydroxyethyl methacrylate, 2-hydroxypropyl methacrylate, or glycidyl methacrylate; further, diethyl or monoesters of unsaturated dicarboxylic acids; further, (meth)acrylamide-N-methylol methyl ether, amide or nitrile, such as acrylamide, methacrylamide, N-methylol(meth)acrylamide, acrylonitrile, methacrylonitrile; and also N-substituted maleimides and ethers, such as vinyl butyl ether, vinyl isobutyl ether, or vinyl phenyl ether, and combinations thereof. In another embodiment, gelling agents that can be used as rheological modifiers or anti-settling agents may include hydrophobic modified clay (e.g., sodium montmorillonite in which exchangeable sodium ions are replaced with organic cation molecules such as alkylamines), surface-modified silica, fumed silica (e.g., untreated or surface-modified), and combinations thereof.Examples of surface-modified fumed silica include fumed silica surface-modified with hexamethyldisilazane, dimethyldichlorosilane (DiMeDi), polydimethylsiloxane, hydrophobic fumed silica post-treated with DDS, and (dimethyldichlorosilane).

[0093] In one embodiment, the composition optionally includes a settling inhibitor / rheological modifier. The rheological modifier may be selected from silica such as fumed silica or treated fumed silica, clay such as bentonite, polyester block copolymer, ethylene / vinyl acetate copolymer, ethylene / acrylic acid copolymer, sorbitol / sebacic acid copolymer behenate, dextrin palmitate, stearoy inulin, thickening oil gel, poly C10-30 alkyl acrylate, styrene and ethylene / propylene diblock copolymer, hydrogenated castor oil, polyamide, or a combination thereof. In a preferred embodiment, the composition includes clay as the rheological modifier. In a preferred embodiment, the composition includes polyester block copolymer as the rheological modifier. In a preferred embodiment, the composition includes fumed silica as the rheological modifier.

[0094] In a preferred embodiment, the composition contains any rheological modifier in the range of about 0.1% w / w to about 20% w / w. In another preferred embodiment, the composition contains any rheological modifier in the range of about 0.5% w / w to about 10% w / w. In yet another preferred embodiment, the composition contains any rheological modifier in the range of about 1% w / w to about 5% w / w. In a more preferred embodiment, the composition contains any rheological modifier in the range of about 1% w / w to about 5% w / w.

[0095] In a more preferred embodiment, the composition comprises fumed silica, which is present in the composition at a concentration of about 0.1% w / w to about 20% w / w. In another preferred embodiment, the composition comprises fumed silica, which is present in the composition at a concentration of about 0.5% w / w to about 10% w / w. In yet another preferred embodiment, the composition comprises fumed silica, which is present in the composition at a concentration of about 1% w / w to about 5% w / w. In a more preferred embodiment, the composition comprises hydrophobic fumed silica (dimethyldichlorosilane), which is hydrophobic fumed silica (dimethyldichlorosilane), which is present in the composition at a concentration of about 1% w / w to about 5% w / w.

[0096] In at least one embodiment, the compositions of the Disclosure may optionally comprise one or more polymers. Alternatively, one or more polymers may be applied simultaneously with or sequentially with the biopesticides disclosed herein. Non-limiting uses of polymers in the agricultural industry include pesticide delivery (e.g., as aqueous dispersants), heavy metal removal, water retention and / or water delivery, and combinations thereof (Pouci, et al, Am. J. Agri. & Biol. Sci., 3(7):299-314 (2008)). In one embodiment, one or more polymers may be natural polymers (e.g., agar, starch, alginate, pectin, cellulose, etc.), synthetic polymers, biodegradable polymers (e.g., polycaprolactone, polylactic acid, polyvinyl alcohol, etc.), or combinations thereof.

[0097] In another embodiment, the composition further comprises any anionic surfactant. Examples include alkyl sulfates, alkyl ether sulfates, alkylamide ether sulfates, alkylaryl polyether sulfates, alkylaryl sulfates, alkylaryl sulfonates, monoglyceride sulfates, alkyl sulfonates, alkylamide sulfonates, alkylaryl sulfonates, benzene sulfonates, toluene sulfonates, xylene sulfonates, cumene sulfonates, alkylbenzene sulfonates, alkyldiphenyl oxide sulfonates, alphaolefin sulfonates, alkylnaphthalene sulfonates, paraffin sulfonates, lignin sulfonates, alkyl sulfosuccinates, ethoxylated sulfosuccinates, alkyl ether sulfosuccinates, alkylamide sulfosuccinates, alkyl sulfosuccinate esters, alkyl sulfoacetates, alkyl phosphates, phosphate esters, alkyl ether phosphates, acyl sarcosinates, acyl isethionates, N-acyl taurates, N-acyl-N-alkyl taurates, and alkyl carboxylates. The anionic surfactant may be selected from linear dodecylbenzenesulfonate calcium salt, polyoxyethylene alkyl ether phosphate, polyoxyethylene branched tridecyl phosphate, or a mixture of alkylarylsulfonate calcium and ethylhexanol. In another embodiment, the anionic surfactant may be present in an amount ranging from about 0.1% w / w to about 20% w / w. In another embodiment, the anionic surfactant may be present in an amount ranging from about 0.5% w / w to about 10% w / w. In another preferred embodiment, the anionic surfactant may be present in an amount ranging from about 1% w / w to about 5% w / w.

[0098] In one embodiment, this disclosure, a) an amount of entomopathogenic fungi or their spores in the range of approximately 0.1% w / w to approximately 30% w / w of the total weight of the composition, b) A vegetable oil in an amount ranging from approximately 10% w / w to approximately 99.8% w / w of the total weight of the composition, c) At least two nonionic surfactants with a hydrophilic-lipophilic balance (HLB) value of 15 or less, in an amount ranging from approximately 0.1% w / w to approximately 60% w / w of the total weight of the composition. The present invention provides a stable, non-aqueous composition containing a nonionic surfactant.

[0099] In one embodiment, this disclosure, a) Beauveria bassiana or its spores, b) Vegetable oil and, c) A combination of a water-insoluble nonionic surfactant and a nonionic polymer surfactant, wherein the HLB values ​​of the water-insoluble nonionic surfactant and the nonionic polymer surfactant are in the range of 4 to 12, The present invention provides a stable, non-aqueous composition containing [the specified element].

[0100] In a preferred embodiment, the non-aqueous composition is an oil dispersion.

[0101] In preferred embodiments, the HLB value of the water-insoluble nonionic surfactant is in the range of 8 to 12.

[0102] In preferred embodiments, the HLB value of the nonionic polymer surfactant is in the range of 4 to 8.

[0103] In a preferred embodiment, the vegetable oil is sunflower oil.

[0104] In preferred embodiments, the nonionic surfactant is selected from polyoxyethylene sorbitol hexaoleate, polyisobutylene succinic anhydride-polyethylene glycol, or a combination thereof. In one embodiment, the HLB values ​​of polyoxyethylene sorbitol hexaoleate and polyisobutylene succinic anhydride-polyethylene glycol are in the range of 4 to 12.

[0105] In one embodiment, the weight ratio of polyoxyethylene sorbitol hexaoleate to polyisobutylene succinic anhydride-polyethylene glycol in the composition is in the range of about 1:10 to about 10:1.

[0106] The compositions of this disclosure can be in any form, regardless of their form (e.g., nutrient state or dormant state), as long as they support the desired activity (effective amount) of a fungal insecticide (entomopathogenic fungus or its spores), and can be applied to control target pests.

[0107] In one embodiment, the present disclosure provides a method for preparing a stable non-aqueous composition, the method being: i) A step of mixing a vegetable oil with at least two nonionic surfactants having a hydrophilic-lipophilic balance (HLB) value of 15 or less under high shear, ii) The step of adding an entomopathogenic fungus or its spores to the mixture obtained in step i), iii) A step of homogenizing the mixture from step ii) to obtain a stable composition, Includes.

[0108] In one embodiment, the method includes adding any rheological modifier or any anionic surfactant under high shear to the mixture obtained in step i) before adding an entomopathogenic fungus or its spores.

[0109] In some embodiments, the composition is prepared as a premixed composition. In some embodiments, the composition is prepared by immediately mixing fungal spores with components including oil, surfactant, and other optional components at the time of application. In another embodiment, the composition is prepared by mixing fungal spores, oil, surfactant, and other optional components substantially before application of the composition to the target site or pest.

[0110] One embodiment of this disclosure is, i) A step of mixing a vegetable oil with at least two nonionic surfactants having a hydrophilic-lipophilic balance (HLB) value of 15 or less under high shear, ii) The step of adding an entomopathogenic fungus or its spores to the mixture obtained in step i), iii) A step of homogenizing the mixture from step ii) to obtain a stable composition, The present invention provides an oil dispersion obtained by a method including the following:

[0111] In one embodiment, the method includes adding any rheological modifier or any anionic surfactant under high shear to the mixture obtained in step i) before adding an entomopathogenic fungus or its spores.

[0112] Entomological fungi or their spores can be produced by liquid or solid medium fermentation processes. The media may have high carbon and nitrogen concentrations to obtain higher yields. Non-limiting examples of suitable nitrogen sources include hydrolyzed casein, yeast extract, hydrolyzed soy protein, hydrolyzed cottonseed protein, and hydrolyzed corn gluten protein. Non-limiting examples of suitable carbon sources include carbohydrates such as glucose, fructose, and sucrose, glycerol, and / or cereals such as rice and barley.

[0113] The fermentation process may be carried out using conventional fermentation processes such as aerobic liquid culture techniques, shaking flask culture, and small-scale or large-scale fermentation in laboratory or industrial fermenters (e.g., continuous, batch, fed-batch, solid-state fermentation, etc.), and such processes are well known in the art. Regardless of the preparation process used to produce the entomopathogenic fungus or its spores, it is assumed that the entomopathogenic fungus or its spores may be used as an insecticide directly from the culture medium or after purification and / or further processing steps (e.g., drying process). In one embodiment, after fermentation, the entomopathogenic fungus or its spores may be recovered using conventional techniques (e.g., filtration, centrifugation, mechanical recovery (e.g., shaking the entomopathogenic fungus or its spores from the culture medium), etc.). The entomopathogenic fungus or its spores may be dried (e.g., air-dried, freeze-dried, or spray-dried to a low moisture level and stored at a suitable temperature, e.g., room temperature).

[0114] The components of the composition (i.e., spores and emulsion) may be provided as a kit and housed in a single container, or preferably in separate containers. Users may choose to mix the components immediately at the time of application to prepare the composition, or they may choose to mix the components well in advance of application to prepare the composition and store it for a period that does not affect the stability and infectivity of the composition until application.

[0115] In one embodiment, this disclosure, a) Entomogenic fungi or their spores, b) Vegetable oil and, c) At least two nonionic surfactants with a hydrophilic-lipophilic balance (HLB) value of 15 or less, The present invention provides a method for improving the storage stability and viability of entomopathogenic fungi or their spores, comprising preparing a non-aqueous composition containing [a specific substance].

[0116] In one embodiment, additional agriculturally beneficial components (e.g., beneficial microorganisms, signaling molecules, insecticides, fungicides, nematicides, and combinations thereof) may also be used in combination with the compositions described herein, either as part of the same composition or as a separate treatment.

[0117] This specification also discloses methods for controlling pests. In one embodiment, this method controls one or more species of plant pests. a) Entomogenic fungi or their spores, b) Vegetable oil and, c) At least two nonionic surfactants with a hydrophilic-lipophilic balance (HLB) value of 15 or less, The process includes the step of bringing the mixture into contact with a composition containing the following:

[0118] In one embodiment, the composition is applied at a rate of approximately 0.1 to 50 L / ha.

[0119] In another embodiment, a method for controlling one or more species of pests using compositions disclosed herein is disclosed. In a preferred embodiment, the method includes the step of controlling one or more species of plant pests. Non-limiting examples of plant pests include:

[0120] Hemiptera pests: Planthoppers such as Laodelphax striatellus, Nilaparvata lugens, and Sogatella furcifera (Delphacidae); leafhoppers such as Nephotettix cincticeps and Nephotettix virescens (Deltocephalidae); cotton aphids (Aphis gossypii), peach aphids (Myzus persicae), cabbage aphids (Brevicoryne brassicae), potato aphids (Macrosiphum euphorbiae), digitalis aphids (Aulacorthum solani), wheat aphids (Rhopalosiphum padi), and citrus aphids (Toxoptera) Aphids such as *citricidus* (Aphididae); Stink bugs such as *Nezara Antennata*, *Riptortus clavetus*, *Leptocorisa chinensis*, *Eysarcoris parvus*, *Halyomorpha mista*, and *Lyus lineolarxs* (Pentatomidae); Whiteflies such as *Trialeurodes Vaporariorum*, *Bemisia tabaci*, and *Bemisia argentifolii* (Aleyrodidae); Silverleaf whitefly (Aonidiella aurantii) and San Jose scale (Comstockaspis) Scales of insects such as *Perniciosa*, *Unaspis citri*, *Ceroplastes rubens*, and *Icerya purchasi* (family Coccinaceae); citrus mealybug (family Chelydidae); and large lice (family Coricidalidae);

[0121] Lepidopteran pests: Moths such as the rice leaf borer (Chilo suppressalis), the red leaf borer (Tryporyza incertulas), the wheat leaf borer (Cnaphalocrocis medinalis), the cotton leaf borer (Notarcha derogata), the Indian meal moth (Plodia interpunctella), the corn leaf borer (Ostrinia furnacalis), the European corn borer (Ostrinia nubilaris), the diamondback moth (Hellula undalis), the sod web moth (Pediasia teterrellus), the beet armyworm (Spodoptera litura), the white-striped armyworm (Spodoptera exigua), the corn armyworm (Pseudaletia separata), the cutworm (Mamestra brassicae), the cabbage moth (Agrotis ipsilon), and the cabbage looper (Plusia Noctuid moths such as *nigrisigna*, *Trichoprucia*, *Heliotis*, and *Helicobelpa*, white butterflies such as *Pieris rapae*, *Adoxophyes*, pear fruit moth (Grapholita molesta), bean fruit moth (Leguminivora glycinivorella), adzuki bean pod moth (Matsumuraeses azukivora), apple leaf roller moth (Adoxophyes orana fasciata), tea leaf roller moth (Adoxophyes sp.), leafrollers such as Homona magnanima, Archips fuscocupreanus, and Cydia pomonella; narrow-legged moths such as Caloptilia theivora and Phyllonorycter ringoneella; fruit moths such as Carposina niponensis; Lyonetiidae moths such as Lyonetia; crested moths such as Lymantriidae moths and Eupractis; cutworms such as Plutella xylostella; gelechiidae moths such as Pectinophora gossypiella and Phthorimaea operculella; and fall webworms (Hyphantria japonensis). This includes moths such as *Cunea* (a type of moth belonging to the Arctiidae family), the European greenfinch (*Tinea translucens*), and jumping spiders (*Tineola bisselliella*), among others.

[0122] Thrips pests: Thrips (Thripidae), such as the orange thrips (Frankliniella occidentalis), melon thrips (Thrips palmi), tea leaf thrips (Scirtothrips dorsalis), onion thrips (Thrips tabaci), orange thrips (Frankliniella intonsa), tobacco thrips (Frankliniella fusca), etc.

[0123] Diptera pests: Houseflies (Musca domestica), house mosquitoes (Culex popiens pallens), horseflies (Tabanus trigonus), onion flies (Hylemya antiqua), seed flies (Hylemya platura), Asian lizards (Anopheles sinensis); leafminers (Agromyzidae), such as rice leafminers (Agromyza oryzae), dwarf rice leafminers (Hydrellia griseola), rice stem flies (Chlorops oryzae), bean leafminers (Liriomyza trifolii), etc.; cucumber flies (Dacus cucurbitae), Mediterranean fruit flies (Ceratitis capitata), etc.

[0124] Coleopteran pests: Epilachna vigintioctopunctata, Cucumber beetle (Aulacophora femoralis), Striped flea beetle (Phyllotreta striolata), Rice leaf beetle (Oulema oryzae), Rice curculio (Echinocnemus squameus), Rice water weevil (Lissorhoptrus oryzophilus), Giant weevil (Anthonomus grandis), Adzuki bean weevil (Callosobruchus chinensis), Hunting beetle (Sphenophorus venatus), Japanese leaf beetle (Popxllia japonica), Copper beetle (Anomala cuprea), Corn root worm (Diabrotica spp.), Colorado leaf beetle (Leptinotarsa ​​decemlineata), Click beetle (Agriotes spp.), Tobacco leaf beetle (Lasioderma) Examples include serricorne, false black tree (Anthrenus verbasci), red dwarf weevil (Tribolium castaneum), powder beetle (Lyctus brunneus), white-spotted longhorn beetle (Anoplophora malasiaca), and Japanese pine sawyer beetle (Tomicus piniperda);

[0125] Orthopteran pests: Examples include the migratory locust (Locusta migratoria), African mole cricket (Gryllotalpa africana), Japanese grasshopper (Oxya yezoensis), and Japanese grasshopper (Oxya japonica);

[0126] Hymenoptera pests: Cabbage sawflies (Athalia rosae), leafcutter ants (Acromyrmex genus), fire ants (Solenopsis genus), etc.

[0127] Cockroach pests: Examples include the German cockroach (Blattella germanica), the American cockroach (Periplaneta fuliginosa), the American cockroach (Periplaneta americana), the American cockroach (Periplaneta brunnea), and the Eastern cockroach (Blatta orientalis);

[0128] Specific examples of the harmful arthropods mentioned above include aphids (Aphididae), thrips (Thripidae), leafminers (Agromidae), horsehair worms (Paragordius tricuspidatus), Colorado leaf beetles (Leptinotarsa ​​decemlineata), Japanese leaf beetles (Popillia japonica), scarab beetles (Anomala cuprea), giant weevils (Anthonomus grandis), rice water weevils (Lissorhoptrus oryzophilus), tobacco thrips (Frankliniella fusca), corn root worms (Diabrotica spp.), diamondback beetles (Plutella xylostella), cabbage weevils, and soybean pod borers (Leguminivora glycinivorella).

[0129] In a preferred embodiment, the method includes controlling one or more species of plant pests, comprising the step of bringing the plant pests into contact with one or more species of the compositions of the Disclosure. The contact step can be carried out by any method known in the art (e.g., spraying, dispensing, etc.). In one embodiment, the contact step is repeated (e.g., two or more times, such as two, three, four, five, six, seven, eight, nine, ten, etc.).

[0130] In another embodiment, a method for controlling one or more species of pests includes the step of bringing the pests or a plant or part of a plant into contact with one or more of the compositions of the Disclosure. While not theoretically bound, it is assumed that one or more species of pests, such as plant pests, will come into contact with the compositions disclosed herein upon contact with the treated plant or part of a plant. In one embodiment, the contact step can be carried out by any method known in the Art (including both foliar and non-foliar application). Non-limiting examples of contacting a plant or part of a plant include spraying, dipping, dripping, spraying, and / or coating seeds with one or more of the compositions of the Disclosure. In one embodiment, the contact step is repeated (e.g., one or more times, such as the treatment step being repeated two, three, four, five, six, seven, eight, nine, ten times, etc.). The contact step can be carried out at any point during the growth of the plant or part of a plant. In one embodiment, the step of bringing a plant or plant part into contact with one or more of the compositions of the Disclosure is performed before the plant or plant part begins to grow. In another embodiment, the step of bringing a plant or plant part into contact with one or more of the compositions of the Disclosure is performed after the plant or plant part has begun to grow.

[0131] In another embodiment, the method further includes a step of treating soil or locus with the compositions of the Disclosure. The planting step can be performed before, after, or during treatment of the soil with one or more of the compositions of the Disclosure. In one embodiment, the planting step is performed before the soil is treated with one or more of the compositions described herein. In another embodiment, the planting step is performed while the soil is being treated with one or more of the compositions of the Disclosure (for example, the planting step is performed substantially simultaneously with the treatment step, etc.). In yet another embodiment, the planting step is performed after the soil has been treated with one or more of the compositions of the Disclosure.

[0132] In another embodiment, seeds are coated with one or more of the compositions of the Disclosure. In one embodiment, seeds may be treated with the compositions described herein in several ways, such as spraying or dripping. Spraying and dripping treatments can be carried out by formulating the compositions of the Disclosure and spraying or dripping the compositions of the Disclosure onto the seeds(s) via a continuous treatment system, such as a drum-type treatment device, which is calibrated to apply the treatment at a predetermined rate proportional to the continuous flow of seeds. A batch system may also be employed, which sends a predetermined batch size of seeds and compositions(s) described herein to a mixer. In another embodiment, the treatment involves coating the seeds. In one embodiment, seeds coated with the compositions of the Disclosure are further disclosed.

[0133] In one embodiment, the composition of the Disclosure may be used for pest control not only in agricultural areas but also in non-agricultural areas. In one embodiment, the composition of the Disclosure may be used for pest control in crops. In one embodiment, the composition of the Disclosure may be used for pest control in non-agricultural plants.

[0134] The compositions of this disclosure may be applied at varying concentrations to carry out any of the methods disclosed or to any of the seed coatings or seed coating methods described herein. In one embodiment, the compositions of this disclosure are diluted with water. In one embodiment, the compositions of this disclosure are applied at a rate of about 0.1 to about 20 kg / ha.

[0135] The present invention is further illustrated in the following embodiments. However, it should be understood that these embodiments are for illustrative purposes only and do not limit the scope of the invention in any way. [Examples]

[0136] Example 1: An oil dispersion composition containing Beauveria bassiana spores according to the present invention is prepared as follows.

[0137] [Table 1]

[0138] This composition was prepared by mixing sunflower oil with two types of nonionic surfactants. Fumed silica was added under high shear and mixed until homogeneous. Beauveria bassiana spores were added to the resulting homogeneous mixture and further homogenized to obtain a stable oil dispersion.

[0139] Example 2: Comparative stability evaluation of oil-based surfactants.

[0140] Eighteen compositions consisting of oil and surfactant were prepared using compositions without active ingredients (excluding compositions 5 and 9), and the stability of these compositions over 15 hours was evaluated. Compositions 1 to 18 are listed in Table 2.

[0141] [Table 2] TIFF2026511053000004.tif231166TIFF2026511053000005.tif230166TIFF2026511053000006.tif224165TIFF2026511053000007.tif126164

[0142] From the table above, it was demonstrated that only compositions containing two nonionic surfactants with HLB values ​​of 15 or less were stable, while other compositions containing anionic surfactants, a single surfactant, and a nonionic surfactant with an HLB value greater than 15 were unstable. Furthermore, compositions 3, 4, 17, and 18 were the most stable compositions, with one nonionic surfactant having an HLB value of less than 8 and the other nonionic surfactants having an HLB value greater than 8. Composition 5 showed good spore dispersion in the system.

[0143] Example 3: The oil dispersion of Beauveria bassiana spores according to the present disclosure is shown in Table 3.

[0144] [Table 3]

[0145] Example 4: The oil dispersion of Beauveria bassiana spores according to the present disclosure is shown in Table 4.

[0146] [Table 4]

[0147] Example 5: Table 5 shows the oil dispersion of Beauveria bassiana spores according to the present disclosure.

[0148] [Table 5]

[0149] Example 6: The oil dispersion of Beauveria bassiana spores according to the present disclosure is shown in Table 6.

[0150] Table 6

Claims

1. A stable non-aqueous composition, (a) Entomogenic fungi or their spores, (b) Vegetable oil and (c) At least two nonionic surfactants with a hydrophilic-lipophilic balance (HLB) value of 15 or less, A stable non-aqueous composition comprising the entomopathogenic fungus or its spores in an amount ranging from about 0.1% w / w to about 30% w / w of the total weight of the composition.

2. The stable non-aqueous composition according to claim 1, wherein the stable non-aqueous composition is an oil dispersion.

3. The stable non-aqueous composition according to claim 1, wherein the entomopathogenic fungus or its spores is Beauveria bassiana or its spores.

4. The stable non-aqueous composition according to claim 1, wherein the vegetable oil is selected from olive oil, kapok oil, castor oil, palm oil, sunflower oil, camellia oil, coconut oil, sesame oil, corn oil, rice bran oil, peanut oil, cottonseed oil, soybean oil, rapeseed oil, linseed oil, tung oil, canola oil, corn oil, or a combination thereof.

5. The stable non-aqueous composition according to claim 1, wherein the vegetable oil is present in an amount ranging from about 10% w / w to about 99.8% w / w of the total weight of the composition.

6. The stable non-aqueous composition according to claim 1, wherein the HLB values ​​of the at least two nonionic surfactants are 12 or less.

7. The stable non-aqueous composition according to claim 6, wherein one of the two nonionic surfactants has an HLB value in the range of 8 to 12, and the other nonionic surfactant has an HLB value in the range of 4 to 8.

8. The stable non-aqueous composition according to claim 1, wherein the weight ratio of the two nonionic surfactants in the composition is in the range of about 1:100 to about 100:

1.

9. The stable non-aqueous composition according to claim 1, wherein the at least two nonionic surfactants are included in an amount ranging from about 0.1% w / w to about 60% w / w of the total weight of the composition.

10. The stable non-aqueous composition according to claim 1, wherein the nonionic surfactant is selected from a water-soluble nonionic surfactant, a water-insoluble nonionic surfactant, a nonionic polymer surfactant, or a combination thereof.

11. The stable non-aqueous composition according to claim 10, wherein the nonionic polymer surfactant is selected from polymer surfactants selected from polyisobutylene succinic anhydride-polyethylene glycol, polyalkylene oxide block copolymer, polyalkylene glycol ether, polyoxyalkylene glycol aliphatic alcohol ether, polyoxyethylene alkyl ether, block copolymer of polyethylene glycol and 12-hydroxystearic acid, polymethyl methacrylate-polyethylene glycol graft copolymer, succinic anhydride-polyethylene glycol, polyoxyethylene polyarylphenol, block copolymer of poly(12-hydroxystearic acid) and poly(ethylene oxide), block copolymer of poly(12-hydroxystearic acid) and poly(ethylene oxide) having a molecular weight of 1500 to 10000 (also known as polyethylene glycol), alkylbenzene sulfonates, alkanoyl taurates, and alkanoyl sarcosinates, and combinations thereof.

12. The stable non-aqueous composition according to claim 10, wherein the water-insoluble nonionic surfactant is selected from alkyl and aryl:glycerol ethers, glycol ethers, ethanolamides, sulfonylamides, alcohols, amides, alcohol ethoxylates, ethoxylated oils such as castor oil ethoxylate, glycerol esters, glycol esters, ethoxylates of glycerol esters and glycol esters, sugar-based alkyl polyglycosides, polyoxyethylene-conjugated fatty acids, alkanolamine condensates, alkanolamides, tertiary acetylene glycols, polyoxyethylene-conjugated mercaptans, carboxylic acid esters, polyoxyethylene-conjugated polyoxypropylene glycols, sorbitan fatty acid esters, sorbitol ethoxylate esters, EO / PO block copolymers (EO is ethylene oxide, PO is propylene oxide), EO polymers and copolymers, polyamines, polyvinylpyrrolidone, sodium sulfosuccinate (such as sodium dioctyl sulfosuccinate), and combinations thereof.

13. The stable non-aqueous composition according to claim 10, wherein the water-soluble nonionic surfactant is selected from sorbitan fatty acid alcohol ethoxylate, alkoxylated diethylethanolamine, sorbitan fatty acid ester ethoxylate, and combinations thereof.

14. The stable non-aqueous composition according to claim 1, further comprising any rheological modifier or any anionic surfactant.

15. The stable non-aqueous composition according to claim 1, wherein the composition is a biopesticide composition.

16. A method for improving the storage stability and viability of the entomopathogenic fungus or its spores, comprising the step of preparing the stable non-aqueous composition described in claim 1.

17. A method for preparing the stable non-aqueous composition described in claim 1, wherein the method is: (i) A step of mixing vegetable oil with at least two nonionic surfactants with an HLB value of 15 or less under high shear, (ii) The step of adding an entomopathogenic fungus or its spores to the mixture obtained in step i), (iii) A step of homogenizing the mixture from step ii) to obtain a stable composition, Methods that include...

18. A method for preparing the stable non-aqueous composition according to claim 17, comprising the step of optionally adding a rheological modifier or an anionic surfactant to the mixture obtained in step i) under high shear, before adding the entomopathogenic fungus or its spores.

19. A method for controlling plant pests, comprising the step of bringing one or more species of plant pests or their habitat into contact with the stable non-aqueous composition described in claim 1.

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