Compositions and methods for reducing still water and controlling aquatic breeding pests

Abstract

The composition provided by the invention is used for treating still water (such as reservoirs and puddles) and soil where flood and ponding are prone to occur. More specifically, the present invention provides microorganism-derived compositions and their use in preventing and / or ameliorating (i.e., discharging) undesired still water and / or problems associated with still water, such as mosquito and algae growth, etc. Advantageously, the compositions and methods of the present invention can be used as environmentally friendly, non-toxic and cost effective solutions for improved moisture utilization efficiency, soil health and pest control.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Patent Application No. 63 / 319,490, filed on March 14, 2022, the contents of which are incorporated herein by reference in their entirety. Background Art

[0003] Soil is a complex complex of minerals, gases, liquids, organic matter, and microorganisms. The exact composition of a particular type of soil varies depending on factors such as human activity, geographic location, and climate. The health of the soil may also depend on these factors. Degradation, compaction, hydrophobicity, and inefficient water and nutrient transport are all examples of problems that can occur in certain soils, each of which can have a negative impact on the environment and negatively affect farms, golf courses and sports fields, lawns, developed areas, and pastures.

[0004] For example, compaction can cause soil particles to be pressed closer together, thereby reducing the pore space between them, due to changes in soil particle distribution, organic matter content, mineral composition, and water content. Well-sorted fine sandy loams and loamy fine sands with high fine sand content and low carbon content are particularly susceptible to compaction.

[0005] Compaction can also be caused by trampling by livestock, the use of heavy machinery, certain irrigation methods, and poorly managed tillage practices. Over time, compaction can lead to degradation of soil physical properties, reduction of microbial populations, reduction of soil organic fraction (SOC) content, and changes in the size, structure, and number of soil pores. Conversely, as bulk density increases, soil strength increases, while water and air conductivity, permeability, and diffusivity decrease. Soil layers with high impermeability reduce rooting depth and root density, resulting in reduced plant uptake of nutrients and water and reduced water use efficiency. In addition, inefficient water and nutrient transport can lead to flooding and ponding (such as standing or stagnant water), as well as runoff of phosphates and nitrates, which can lead to harmful algal blooms.

[0006] Soil hydrophobicity is another problem that can occur in soils. Soils themselves may be naturally hydrophobic, a property that tends to be exacerbated when soils are dry for long periods of time or have high organic matter content. Fires can also cause or exacerbate soil hydrophobicity as hydrophobic materials in soils and debris volatilize, pyrolyze, and redistribute deeper into the soil profile.

[0007] Soil hydrophobicity can limit irrigation-based applications as well as rainwater infiltration and infiltration. As a result, soil hydrophobicity can also lead to flooding, ponding, evaporation, and surface runoff, which directly affects water use efficiency and plant growth by limiting water infiltration and supply to the plant root zone. In addition, ponding can lead to inoculation and growth of undesirable life forms such as algae and mosquitoes.

[0008] Mosquitoes, in particular, are not only a nuisance as biting insects, but they also have the potential to cause other harmful and widespread consequences. Mosquitoes are vectors of a wide range of diseases that can have serious effects on humans, pets, and livestock. For example, mosquitoes of the genus Aedes are responsible for transmitting several dangerous human viruses, including those that cause malaria, dengue, yellow fever, chikungunya, and most recently, Zika.

[0009] The areas most conducive to mosquito populations are those with hot, humid climates, although mosquitoes can survive in almost any habitat with plenty of stagnant water. This is because key stages of the mosquito life cycle are primarily completed in water. Female mosquitoes of container-breeding mosquito species lay their eggs on the surface of water in man-made containers, puddles, swamps, or small pools. Other mosquitoes lay their eggs in moist or dry soil, waiting to be submerged by rising water. Once the eggs hatch in the water, the young mosquitoes remain in the water for 9 to 14 days, undergoing larval and pupal development stages. Only after the mosquitoes develop into adults do they finally leave the water.

[0010] Due to the potential dangers posed by mosquito bites, monitoring, controlling, and / or modifying the behavior of mosquito populations is essential to prevent the widespread spread of mosquito-borne diseases. For example, draining or preventing standing water can help eliminate mosquito breeding areas. Additionally, mosquito populations can be effectively reduced by introducing traps, protecting natural predators or enemies, and spreading insecticides in mosquito habitats. However, widespread use of insecticides can result in the long-term presence of toxic chemicals in the environment and have adverse effects on beneficial species.

[0011] The economic costs and environmental impacts of poor soil health and inefficient water use are a persistent burden for farmers, gardeners, homeowners and pastoralists. The risk of mosquito-borne disease is another factor, so improved, safe approaches to soil and water management are needed, especially in climates where mosquitoes thrive. Summary of the invention

[0012] The present invention provides compositions and methods for treating standing water (such as reservoirs and puddles) and soils that are prone to flooding and ponding. More specifically, the present invention provides microbial-derived compositions and their use in preventing and / or mitigating (i.e., discharging) undesirable standing water and / or problems associated with standing water (such as mosquitoes and algae growth, etc.). Advantageously, the compositions and methods of the present invention can be used as environmentally friendly, non-toxic and cost-effective solutions for improving water use efficiency, enhancing soil health and controlling pests.

[0013] In certain embodiments, the present invention provides a water treatment composition comprising a surface active molecule. The water treatment composition can be used for a variety of purposes, including:

[0014] a) wetting agents, which are used to promote the drainage of accumulated water into the soil;

[0015] b) preventive treatments used to enhance the wettability and drainage of soils prior to contact with water, thereby preventing ponding and flooding; and

[0016] c) A pest control agent for controlling mosquitoes, mosquito larvae and / or algae in stagnant water.

[0017] Thus, in certain embodiments, methods are provided for applying water treatment to natural and artificial sources of standing water and / or soil. In some embodiments, the method comprises applying the composition to water accumulated on soil, a water container such as a sink, or a body of water such as a pond. In some embodiments, the method comprises applying the composition to the soil before the soil is contacted with irrigation water or rainfall.

[0018] Advantageously, in addition to reducing standing water and / or controlling pests, in some embodiments, the methods and compositions are effective in increasing water use efficiency and soil flow rates under a variety of conditions, including, for example, compacted soils, drought soils, eroded soils, nutrient-depleted soils, waterlogged soils, fire-damaged soils, hydrophobic soils, and / or soils with plants growing therein.

[0019] In a preferred embodiment, the surfactant molecules of the present invention are microbially derived biosurfactants. The biosurfactants can be applied in purified and / or crude form. The biosurfactants in crude form can include biosurfactants and other cell growth products (including fermentation media generated by culturing microorganisms that generate biosurfactants), etc.

[0020] The biosurfactant according to the method of the present invention can be selected from low molecular weight glycolipids (such as sophorolipids, cellobiose lipids, rhamnolipids, mannoerythritol lipids and trehalose lipids, etc.), lipopeptides (such as surfactin, iturin, fengycin, actin and lichenin, etc.), flavin lipids, phospholipids (such as cardiolipin, etc.), fatty acid esters and high molecular weight polymers (for example, lipoproteins, lipopolysaccharide-protein complexes and polysaccharide-protein-fatty acid complexes), etc.

[0021] In certain specific embodiments, the biosurfactant is a sophorolipid (SLP), such as a lactone SLP, an acidic SLP, an amino acid SLP conjugate, a salt-type SLP, or a derivative of any of these.

[0022] In certain embodiments, the water treatment composition may include additional substances, such as carriers, microbial inoculants, pH adjusters, additional pesticides, herbicides, fertilizers, mineral sources, plant seeds, dyes, stabilizers, emulsifiers, prebiotics, and / or polymers.

[0023] The composition can be applied in combination with an aqueous fluid and applied to water and / or soil. In addition, or as an alternative, the composition can be applied as a dry form of powder or granules, which activate the surface active molecules in the composition when mixed with water.

[0024] In certain embodiments, the water treatment composition, due to its nanoparticle micelle size, is particularly useful for reducing the surface and / or interfacial tension between water and soil, and increasing the porosity of compacted soil. The ultra-small micelle size of the irrigation additive allows water to penetrate into the micro- and nano-pores in hard and compacted soils, thereby opening the pores and thus helping to enhance the mobility of air, nutrients, and water.

[0025] In certain embodiments, the water treatment composition reduces the surface tension of water, thereby controlling larval mosquitoes and causing adult mosquitoes to sink.

[0026] In certain embodiments, the water treatment compositions have biocidal properties, thereby helping to control algae growth.

[0027] The compositions and methods of the present invention can be used in areas such as farms, pastures, fields, lawns, gardens, golf courses, parks, swamps, ditches, drains, and anywhere prone to flooding or standing water, with open containers that collect water, or where standing or stagnant water exists. Advantageously, the compositions and methods of the present invention provide a "green" alternative for pest control while improving water use efficiency and soil health. DETAILED DESCRIPTION

[0028] The present invention provides microbial derived compositions and methods for use thereof to prevent and / or mitigate (such as discharge, etc.) undesirable standing water and / or standing water-related problems (e.g., mosquitoes and algae growth). Advantageously, the compositions and methods of the present invention can be used as environmentally friendly, non-toxic and cost-effective solutions for improving water use efficiency, enhancing soil health and controlling pests.

[0029] Selected Definitions

[0030] As used herein, "agriculture" refers to the cultivation and propagation of plants for food, fiber, biofuels, pharmaceuticals, cosmetics, supplements, ornamental purposes and other uses. According to the present invention, agriculture can also include gardening, landscaping, gardening, plant protection, forestry and reforestation, pasture and grassland restoration, orchards, arboriculture and agronomy. Agriculture also includes the care, monitoring and maintenance of soil.

[0031] As used herein, "broth", "culture broth" or "fermentation broth" refers to a culture medium containing at least nutrients and microbial cells.

[0032] As used herein, the term "carbon utilization efficiency" or "CUE" refers to a broad measure of the efficiency with which a microorganism allocates carbon for growth and biomass production versus respiration. CUE can be calculated as growth (biomass production) divided by the sum of CO2 production / emissions and growth. Microorganisms are generally classified as "low CUE" or "high CUE," where a CUE greater than 0.50 is considered high CUE and a CUE below 0.50 is considered low CUE.

[0033] Unless the context requires otherwise, the phrases "fermentation," "fermentation process," or "fermentation reaction" and the like as used herein are intended to encompass both the growth phase and the product biosynthesis phase of the process.

[0034] As used herein, an "isolated" or "purified" compound is substantially free of other compounds with which it is associated in nature, such as cellular material. A purified or isolated polynucleotide (ribonucleic acid (RNA) or deoxyribonucleic acid (DNA)) is free of the flanking genes or sequences found in its naturally occurring state. A purified or isolated polypeptide is free of the amino acids or sequences that flank it in its naturally occurring state. In the context of microbial strains, "isolated" means that the strain is removed from the natural environment in which it occurs. Thus, an isolated strain may exist as a biologically pure culture or as a spore (or other form of strain) associated with a vector.

[0035] As used herein, a "biologically pure culture" is a culture that has been separated from the material with which it is associated in nature. In preferred embodiments, the culture has been separated from all other living cells. In further preferred embodiments, the biologically pure culture exhibits superior properties compared to cultures of the same microorganism found in nature. The advantageous property may be an increased production of one or more growth byproducts.

[0036] In certain embodiments, the purified compound is at least 60% by weight of the compound of interest. Preferably, the preparation is at least 75%, more preferably at least 90%, and most preferably at least 99% by weight of the compound of interest. For example, the purified compound is at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 98%, 99% or 100% (w / w) of the desired compound by weight. Purity is measured by any appropriate standard method such as column chromatography, thin layer chromatography or high performance liquid chromatography (HPLC) analytical methods.

[0037] As used herein, the terms "enhance," "enhance," and "increase" are used interchangeably.

[0038] "Metabolite" refers to any substance generated by metabolism (e.g., a growth byproduct) or a substance necessary to participate in a specific metabolic process. A metabolite can be an organic compound that is a starting material, intermediate, or end product of metabolism. Examples of metabolites include, but are not limited to, biosurfactants, biopolymers, enzymes, acids, solvents, alcohols, proteins, vitamins, minerals, trace elements, and amino acids.

[0039] The present invention utilizes "microbial-based compositions" to refer to compositions that include components generated by the growth of microorganisms or other cell cultures. Therefore, microbial-based compositions can include the microorganisms themselves and / or byproducts of microbial growth. The microorganisms can be in an asexual state, in the form of spores or conidia, in the form of hyphae, in any other form of microbial propagules, or a mixture thereof. The microorganisms can be planktonic microorganisms or microorganisms in the form of biofilms, or a mixture of the two. The byproducts of growth can be, for example, metabolites, cell membrane components, proteins, and / or other cell components. The microorganisms can be intact microorganisms or lysed microorganisms. In a preferred embodiment, the microorganisms are present in the microbial-based composition together with the growth medium in which they grow. The microorganisms can be present in at least 1x10 per gram or per milliliter of the composition. 4 , 1x10 5 , 1x10 6 , 1x10 7 , 1x10 8 , 1x10 9 , 1x10 10 , 1x10 11 , 1x10 12 or 1x10 13 or more CFU concentrations.

[0040] The present invention further provides "microorganism-based products", which are products that achieve the desired effect in practical applications. Microorganism-based products can simply be microorganism-based compositions obtained from microbial culture processes. Alternatively, microorganism-based products may include other ingredients that have been added. For example, these additional ingredients may include stabilizers, buffers, suitable carriers (such as water, saline solutions or any other appropriate carriers), nutrients added to support further growth of microorganisms, non-nutritional growth promoters and / or agents that help track microorganisms and / or compositions in their application environments. Microorganism-based products may also include mixtures based on microorganism-based compositions. Microorganism-based products may also include one or more components of microorganism-based compositions, which have been processed in some way, such as but not limited to filtration, centrifugation, lysis, drying, purification, etc.

[0041] As used herein, the term "plant" includes, but is not limited to, any kind of woody plants, ornamental or decorative plants, crops or cereals, fruit or vegetable plants, flowers or trees, macroalgae or microalgae, phytoplankton and photosynthetic algae (such as green algae - Chlamydomonas reinhardtii, etc.). "Plants" also include single-cell plants (such as microalgae, etc.) and a large number of plant cells that differentiate into colonies (such as Volvox, etc.) or structures present in any stage of plant development. Such structures include, but are not limited to, fruits, seeds, twigs, stems, leaves, roots, petals, etc. Plants can be grown alone, such as in a garden, or as part of a variety of plants (e.g., part of an orchard, crop, or pasture).

[0042] As used herein, "crop plants" refers to any species of plants or algae grown for profit and / or livelihood for humans, animals or aquatic organisms, or for human use (such as textiles, cosmetics and / or pharmaceutical production, etc.), or for people to enjoy (such as flowers or shrubs in landscaping or gardens, etc.). "Crop plants" also include any plant or algae or part thereof used for industrial, commercial or educational purposes. Crop plants, including transgenic plants and plant varieties, can be obtained by traditional breeding and optimization methods, biotechnology, genetic recombination methods, or a combination of these methods.

[0043] All plants and plant parts can benefit from the present invention. In this case, plants are understood as meaning all plants and plant populations, such as desired and undesired wild plants or crop plants (including naturally occurring crop plants).

[0044] Plant tissue and / or plant parts are understood to mean all above-ground and underground parts and organs of the plant, such as buds, leaves, flowers, roots, needles, stems, stalks, fruits, seeds, tubers and rhizomes. Plant parts also include crop material and vegetative and sexual propagation material, such as cuttings, tubers, rhizomes, young branches and seeds.

[0045] As used herein, "preventing" or "prevention" of a situation or event means delaying, inhibiting, stopping, preventing and / or minimizing the occurrence, spread or progression of the situation or event. Prevention may include but does not require indefinite, absolute or complete prevention, i.e., the situation may still have the potential to occur at a later time. Prevention may include reducing the severity of the situation or event and / or preventing it from further developing into a more serious or more extensive situation or event.

[0046] The range provided herein can be understood as a shorthand expression of all numerical values ​​in the range. For example, the range of 1 to 20 can be understood as any number, combination of numbers or subrange in the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, and all decimal values ​​between the above integers, such as 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8 and 1.9. Regarding subranges, "nested subranges" extending from either end of the range are particularly contemplated. For example, the nested subranges of the exemplary range 1 to 50 can include 1 to 10, 1 to 20, 1 to 30 and 1 to 40 in one direction, or 50 to 40, 50 to 30, 50 to 20 and 50 to 10 in another direction.

[0047] As used herein, "decrease" refers to a negative change and the term "increase" refers to a positive change, wherein the negative change or the positive change is at least 0.25%, 0.5%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 70%, 75%, 80%, 85%, 90%, 95% or 100%.

[0048] As used herein, "reference" refers to a standard or control condition.

[0049] As used herein, "soil amendment" or "soil conditioner" refers to any compound, material, or combination of compounds or materials added to the soil to enhance the soil and / or rhizosphere properties. Soil amendments can include organic and inorganic substances, and may also include fertilizers, pesticides and / or herbicides, etc. Nutrient-rich, well-drained soil is essential for the growth and health of plants. Therefore, soil amendments can increase plant biomass by changing the nutrient and moisture content of the soil. Soil amendments can also be used to improve a variety of different qualities of the soil, including but not limited to soil structure (e.g., preventing compaction); increasing nutrient concentration and storage capacity; increasing water retention in dry soils; and improving drainage in waterlogged soils.

[0050] As used herein, "surfactant" refers to a compound that reduces the surface tension (or interfacial tension) between phases. Surfactants are used as detergents, wetting agents, emulsifiers, foaming agents, and dispersants, etc. "Biosurfactants" are surfactants produced by organisms.

[0051] The transitional term "comprising," which is synonymous with "including" or "containing," is inclusive or open-ended and does not exclude additional, unrecited elements or method steps. In contrast, the transitional phrase "consisting of" excludes any element, step, or ingredient not specified in the claim. The transitional phrase "consisting essentially of" limits the scope of the claim to the specified materials or steps "and those that do not materially affect the basic, novel characteristic." Use of the term "comprising" refers to other embodiments consisting of "consisting of" or "consisting essentially of" the recited components.

[0052] Unless otherwise specified or obvious from the context, the term "or" as used herein should be understood to be inclusive. Unless otherwise specified or obvious from the context, as used herein, the terms "a", "and" and "the" are understood to be singular or plural.

[0053] Unless otherwise specified or apparent from the context, as used herein, the term "about" should be understood to be within the normal tolerance range in the art, such as within 2 standard deviations of the mean. "About" can be understood to be within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05% or 0.01% of the specified value.

[0054] The narration of a list of chemical groups in any definition of a variable herein includes the definition of the variable as any single group or combination of the listed groups. The narration of an embodiment for a variable or aspect herein includes the embodiment as any single embodiment or in combination with any other embodiment or portion thereof. The entire contents of all references cited herein are hereby incorporated herein by reference.

[0055] Compositions and methods for reducing standing water and controlling pests

[0056] In certain embodiments, the present invention provides a water treatment composition comprising a surface active molecule. The water treatment composition can be used for a variety of purposes, including:

[0057] d) wetting agents, which are used to promote the drainage of accumulated water into the soil;

[0058] e) preventive treatments used to enhance the wettability and drainage of soils prior to contact with water, thereby preventing ponding and flooding; and

[0059] f) A pest control agent for controlling mosquitoes, mosquito larvae and / or algae in stagnant water.

[0060] Thus, in certain embodiments, methods are provided wherein the water treatment is applied to standing water and / or soil.

[0061] The compositions and methods of the present invention can be used in areas such as farms, pastures, fields, lawns, gardens, golf courses, parks, swamps, ditches, drains, and any area prone to flooding or standing water, with open containers that collect water, or where standing or stagnant water exists. In certain embodiments, in addition to reducing standing water and / or controlling pests, the methods and compositions are effective in increasing water use efficiency and soil irrigation rates under a variety of conditions, including, for example, compacted soils, drought soils, eroded soils, nutrient-depleted soils, waterlogged soils, fire-damaged soils, hydrophobic soils, and / or soils with plants growing therein.

[0062] In a preferred embodiment, the surfactant molecules of the present invention are microbially derived biosurfactants. The biosurfactants can be applied in purified and / or crude form. The biosurfactants in crude form can include biosurfactants and other cell growth products (including fermentation media generated by culturing microorganisms that generate biosurfactants), etc.

[0063] Biosurfactants are important secondary metabolites produced by various microorganisms such as bacteria, fungi and yeast. As amphiphilic molecules, microbial biosurfactants reduce the surface tension and interfacial tension between liquid, solid and gas molecules. In addition, the biosurfactants according to the present invention are biodegradable biosurfactants, have low toxicity, can effectively dissolve and degrade insoluble compounds in the soil, and can be produced using low cost and renewable resources. They can inhibit the adhesion of undesirable microorganisms to various surfaces, prevent the formation of biofilms, destroy microbial cell walls, and have powerful emulsification and demulsification properties. In addition, biosurfactants can also be used to improve wettability, and achieve uniform solubility and / or distribution of fertilizers, nutrients and water in the soil.

[0064] The biosurfactant according to the method of the present invention can be selected from low molecular weight glycolipids (such as sophorolipids, cellobiose lipids, rhamnolipids, mannoerythritol lipids and trehalose lipids, etc.), lipopeptides (such as surfactin, iturin, fengycin, actin and lichenin, etc.), flavin lipids, phospholipids (such as cardiolipin, etc.), fatty acid esters and high molecular weight polymers (for example, lipoproteins, lipopolysaccharide-protein complexes and polysaccharide-protein-fatty acid complexes), etc.

[0065] In certain specific embodiments, the biosurfactant is a sophorolipid (SLP), such as a lactone SLP, an acidic SLP, an amino acid SLP conjugate, a salt-type SLP, or a derivative of any of these.

[0066] As used herein, the terms "sophorolipid", "sophorolipid molecule", "SLP" or "SLP molecule" include all forms of SLP molecules and isomers thereof, including, for example, acidic (straight chain) SLP (ASL) and lactose SLP (LSL), etc. The terms "sophorolipid", "sophorolipid molecule", "SLP" or "SLP molecule" further include monoacetylated SLP, diacetylated SLP, esterified SLP, SLP with different hydrophobic chain lengths, cationic SLP and / or anionic SLP linked to a fatty acid-amino acid complex, esterified SLP, SLP metal complex, salt-type SLP, SLP amino alcohol, SLP with carbonyl removed from the fatty chain, and other SLPs, including SLPs described and / or not described in the present disclosure.

[0067] In a preferred embodiment, the SLP molecule according to the present invention is represented by the general formula (1) and / or the general formula (2), and comprises fatty acids with different chain lengths (R 3 ) and, in some cases, in R 1 and / or R 2 Acetylation or protonation occurs at the site.

[0068]

[0069] In the general formula (1) or (2), R 0 It can be a hydrogen atom or a methyl group. 1 and R 2 Each of R and R can be independently a hydrogen atom or an acetyl group. 3 It may be, for example, a saturated aliphatic hydrocarbon chain, or an unsaturated aliphatic hydrocarbon chain having at least one double bond, and may have one or more substituents, and the like.

[0070] Non-limiting examples of the substituent include halogen atoms, hydroxyl groups, lower (C1-C6) alkyl groups, halogen-lower (C1-C6) alkyl groups, hydroxy-lower (C1-C6) alkyl groups, halogen-lower (C1-C6) alkoxy groups, and the like. 3 Typically, it will have from 9 to 20 carbon atoms.

[0071] SLPs are typically produced by yeast, such as Candida and / or Candida, such as Candida bombus (Torulopsis), Candida mellifera, Candida batista, Candida floridula, Candida reesii, Candida asteroidea, and / or Candida quinata. SLPs have environmental compatibility, high biodegradability, low toxicity, high selectivity, and specific activity over a wide range of temperature, pH, and salinity conditions.

[0072] In preferred embodiments, the surfactant of the present composition has a micelle size of less than 100 nm, less than 75 nm, less than 50 nm, more preferably less than 25 nm. In certain embodiments, the micelle size is less than 10 nm, less than 8 nm or less than 5 nm.

[0073] In certain embodiments, surface active molecules having a small micelle size (e.g., less than 20 nm) or more preferably an ultra-small micelle size (e.g., less than 10 nm) are particularly helpful in reducing the surface tension and / or interfacial tension between water and soil and increasing the porosity of compacted soil due to their nanoparticle micelle size. The ultra-small micelle size of the irrigation additive allows water to penetrate into the micro- and nano-pores in hard and compacted soils, thereby opening the pores and thus helping to enhance the mobility of air, nutrients, and water.

[0074] In certain embodiments, the perfusion additive may include additional substances such as carriers, pH adjusters, pesticides, herbicides, fertilizers, microbial inoculants, mineral sources, soil amendments, soil conditioners, plant seeds, dyes, stabilizers, emulsifiers, prebiotics and / or polymers.

[0075] As used herein, "applying" a composition or product to a site refers to contacting the composition or product with the site so that the composition or product can have an effect on the site. The mode of application depends on the formulation of the composition and may include spraying, pouring, sprinkling, injecting, spreading, mixing, soaking, atomizing and spraying, etc. The composition of the present invention can be formulated into liquids, dry powders and / or wettable powders, powders, dusts, granules, pellets, emulsions, microcapsules, steaks, oils, gels, pastes and / or aerosols, etc.

[0076] In some embodiments, the method includes applying a water treatment composition to water accumulated on top of the soil to improve drainage of the water into the soil.

[0077] In some embodiments, the method comprises applying the composition to the soil prior to contacting the soil with irrigation water or rainfall.

[0078] In some embodiments, the method comprises applying the composition to a container of standing water, such as a tank, or a body of standing water, such as a pond or puddle formed on soil or a non-soil surface.

[0079] The composition may be premixed with an irrigation fluid, and / or the composition may be applied to a body of water, aqueous or non-aqueous, and / or to the soil surface, wherein the beneficial effects of soil application may be activated by rainfall, sprinkler irrigation, flooding, drip irrigation or other forms of irrigation.

[0080] In one embodiment, the method may include applying the composition to an irrigation system for supplying water, fertilizer, pesticide or other liquid composition. Thus, the composition may be applied by, for example, soil irrigation, soil application, using a center pivot irrigation system, sprayers, micro nozzles, rain sprayers, arm sprayers, sprinklers and / or drippers. Advantageously, the method is suitable for treating hundreds of acres or more of land.

[0081] In one embodiment, wherein the method is used in a smaller scale setting, the method may include spraying the composition onto a site using a hand-held lawn and garden sprayer, or simply pouring the composition into the soil or water using a container or watering can.

[0082] Advantageously, in certain embodiments, the water treatment composition helps to reduce the surface and / or interfacial tension between water and soil contents, thereby increasing the dispersion, permeability and / or infiltration of water and nutrients into the soil, reducing the accumulation and flooding of water on the surface and within the soil, thereby reducing evaporation, runoff and waterlogging.

[0083] In addition, in certain embodiments, the water treatment composition helps loosen hard or compacted soil by penetrating tight soil pores, thereby increasing soil porosity and aeration. Advantageously, this can increase root growth space and water retention within the soil.

[0084] Additionally, in certain embodiments, the water treatment composition reduces the surface tension of water, thereby controlling larval and adult mosquitoes by causing them to sink. Still further, in certain embodiments, the water treatment composition has biocidal properties against, for example, blue-green algae, thereby helping to control algae growth.

[0085] In certain embodiments, the types of mosquitoes that can be controlled include, but are not limited to, the following genera: Lepidopoda, Aedes, Anopheles, Aedes, Aedes, Borachinda, Culex, Echinops, Heterostys, Prolegni, Felis, Trichodina, Aedes aegypti, Colletotrichum, Holothuria, Isostomyia, Isostomoyia, Johnsbelkinia, Chrysophora, Pyralids, Lumos, Gnaphalium, Onirion, Trichopoda, Stracipeda, Stomoxys, Lumos, Gnaphalium, Onirion, Trichopoda, Stomoxys, Lumos, Lumos, Echinops, Hippolytus, Hippolytus, Gnaphalium, Gnaphalium, Prothophobia, Gnaphalium, Gnaphalium, Gnaphalium, Gnaphalium, Gnaphalium, and Gnaphalium. In certain preferred embodiments, the mosquito belongs to the genus Culex (Culex pipiens), the genus Aedes (e.g., Aedes aegypti, Asian tiger mosquito, Aedes africanus, Aedes australis, Aedes ruralis, Aedes polynesiensis), and / or the genus Anopheles (e.g., Anopheles gambiae, Anopheles unguiculata, Anopheles arabiensis). In certain embodiments, the composition can be used against other biting insects that breed in standing water (such as midges).

[0086] In some embodiments, the water treatment composition controls adult mosquitoes that land on standing water to lay eggs, etc. The surface tension of the water is reduced, causing the mosquitoes to fall below the water surface and drown.

[0087] In some embodiments, the water treatment composition affects larval mosquitoes that live below the surface. Both larvae and pupae frequently swim to the surface to breathe, and pupae eventually develop into adults and emerge from the surface. However, in some embodiments, reduced surface tension prevents these organisms from easily moving in and out of the water.

[0088] In some embodiments, the water treatment composition can be applied to the surface of mud or empty containers. Some mosquitoes lay eggs in the mud or on the edges of containers so that when it rains or the water level rises, the eggs become submerged and hatch. Therefore, in certain embodiments, by preventing water from pooling in the mud, or by applying the composition to empty containers that may be filled with water, the composition can help prevent the attachment and / or hatching of eggs.

[0089] In some embodiments, certain species lay eggs in groups or flocks. In some embodiments, the composition can destabilize the attachment of eggs to each other or other surfaces, thereby reducing any protective advantage that exists from grouping.

[0090] In certain embodiments, the concentration of the surfactant is about 0.001% to about 50%, about 0.01% to about 25%, about 0.5% to about 15%, about 2% to about 12%, about 3% to about 10% by weight, or any range therein, compared to the total amount of perfusate administered.

[0091] In certain embodiments, the concentration of the surfactant is about 1 ppm to 1,000 ppm, about 5 ppm to 500 ppm, or about 10 ppm to about 100 ppm compared to the amount of water or perfusate.

[0092] The pH value of the composition should be suitable for the water and soil environment in which it is applied. In some embodiments, the pH is about 2.0 to about 10.0, about 2.0 to about 9.5, about 2.0 to about 9.0, about 2.0 to about 8.5, about 2.0 to about 8.0, about 2.0 to about 7.5, about 2.0 to about 7.0, about 3.0 to about 7.5, about 4.0 to about 7.5, about 5.0 to about 7.5, about 5.5 to about 7.0, about 6.5 to about 7.5, about 3.0 to about 5.5, about 3.25 to about 4.0, or about 3.5. Buffers and pH adjusters (such as carbonates and phosphates) can be used to stabilize the pH near the preferred value.

[0093] In certain embodiments, the compositions and methods can be used to enhance the effectiveness and / or reduce the dosage of other compounds by enhancing the penetration of the pesticidal compound to plants or pests, enhancing the dispersibility of the compound, or increasing the bioavailability of water or nutrients to plant roots, etc. Microbial-based products can also be used to supplement other treatment methods, such as pesticide treatments.

[0094] The methods and compositions of the present invention can be used alone or in combination with other compounds to effectively reduce standing water and control pests associated therewith. For example, in some embodiments, the method includes applying additional components (such as herbicides, fertilizers, pesticides and / or other soil conditioners, etc.) together with the water treatment composition. For example, a grower, pedologist, or entomologist benefiting from the present disclosure can determine the exact materials and their quantities. In some embodiments, the composition can be used as an adjuvant for fertilizers, pesticides, or herbicides.

[0095] In certain embodiments, the methods of the present invention may be further enhanced by applying one or more microbial inoculants. In certain embodiments, the microbial inoculants include one or more soil colonizing microorganisms and / or their growth byproducts, such as biosurfactants, enzymes and / or other metabolites. The inoculant composition may also include a fermentation medium in which the microorganisms are generated.

[0096] In certain embodiments, the microorganism is a bacterium, a yeast, and / or a fungus. In some embodiments, the composition includes more than one type and / or species of microorganism.

[0097] In one embodiment, the microorganism is a Bacillus bacterium, such as Bacillus mojave, Bacillus licheniformis, Bacillus amyloliquefaciens, Bacillus amyloliquefaciens NRRL B-67928, Bacillus subtilis, or Bacillus subtilis NR RL B-68031.

[0098] In one embodiment, the microorganism is a yeast or fungus, such as Trichoderma harzianum, Wickhamella anomala (such as NRRL Y-68030, etc.), Saccharomyces spp. (such as Saccharomyces spp. MEC14XN, Saccharomyces guillemoensis, Saccharomyces calibiker, Saccharomyces calibiker subsp. locus, etc.), Saccharomyces boulardii, Saccharomyces hansdei, Pichia pastoris, and / or Pichia kudriavzevii.

[0099] Advantageously, in some embodiments, the microorganisms can colonize soil and plant roots and help disperse water and salts throughout the soil, control and / or defeat pests, solubilize nutrients for uptake by plant roots, and / or increase above- and below-ground plant biomass compared to untreated soil and / or plants.

[0100] In one embodiment, the microorganism is Bacillus amyloliquefaciens NRRL B-67928 "B.amy". Cultures of the Bacillus amyloliquefaciens "B.amy" microorganism have been deposited at the Northern Regional Research Laboratory (NRRL) Culture Collection of the Agricultural Research Service, located at 1815 N. University St., Peoria, IL, USA. The depository has assigned it a deposit number, NRRL B-67928, and deposited it on February 26, 2020.

[0101] In one embodiment, the microorganism is Bacillus subtilis NRRL B-68031 "B4". A culture of the B4 microorganism has been deposited at the Northern Regional Research Laboratory (NRRL) Culture Collection of the Agricultural Research Service, 1815 N. University St., Peoria, IL, USA. The depository has assigned it the accession number NRRL B-68031 and deposited it on May 6, 2021.

[0102] In one embodiment, the microorganism is Wickham's yeast NRRL Y-68030. Cultures of this microorganism have been deposited at the Northern Regional Research Laboratory (NRRL) Culture Collection of the Agricultural Research Service, located at 1815 N. University St., Peoria, IL, USA. The depository has assigned it the accession number NRRL Y-68030 and deposited it on May 6, 2021.

[0103] Each of the cultures of the invention is stored in conditions that ensure that it is available to persons who, during the life of this patent application, are determined by the Director of the Patent and Trademark Office to be entitled to receive it pursuant to 37 CFR § 1.14 and 35 U.S.C. § 122. The deposit is available in countries where the counterpart of this application or its progeny is filed, as required by foreign patent laws. However, it should be understood that the provision of a deposit does not constitute a derogation from patent rights granted by governmental action, but rather constitutes a license to practice the invention.

[0104] Furthermore, the deposited cultures are maintained and made available to the public in accordance with the provisions of the Budapest Treaty on the Deposit of Microorganisms, i.e., all necessary measures are taken to preserve their viability and protect them from contamination for a period of at least five years after the last request for samples of the deposited cultures and, in any case, for at least 30 (thirty) years after the date of deposit or for the enforceability of any patent that may issue disclosing the cultures. The depositor acknowledges that it is responsible for replacing the deposited cultures if the depositor is unable to provide samples upon request due to the state of the deposit. Upon the grant of a patent disclosing the deposited cultures, all restrictions on making the cultures available to the public are irrevocably lifted.

[0105] The microorganisms and microorganism-based compositions of the present invention have many beneficial properties that can be used to increase plant biomass and / or form / stabilize carbon mineral soil aggregates, etc. For example, the composition can include products produced by the growth of the microorganisms, such as biosurfactants, proteins and / or enzymes in purified or crude form. In addition, the microorganisms can promote plant growth, induce auxin production, solubilize, absorb and / or balance nutrients in the soil, and protect plants from pests and pathogens.

[0106] In a specific embodiment, the concentration of each microorganism included in the inoculant composition is at least 1×10 6 CFU / g to 1x10 13 CFU / g, 1x10 7 CFU / g to 1x10 12 CFU / g, 1x10 8 CFU / g to 1x10 11 CFU / g or 1x10 9 CFU / g to 1x10 10 CFU / g.

[0107] In one embodiment, the total microbial cell concentration of the inoculant composition is at least 1 x 10 6 CFU / g, including up to 1x10 9 CFU / g, 1x10 10 CFU / g, 1x10 11 CFU / g, 1x10 12CFU / g and / or 1x10 13 CFU / g or more CFU / g. In one embodiment, the microorganisms of the inoculant composition comprise about 5% to 100%, or about 8% to 50%, or about 10% to 25% by weight of the total composition weight.

[0108] The inoculant composition may include leftover fermentation substrate and / or purified or unpurified growth byproducts, such as enzymes, biosurfactants and / or other metabolites.The microorganism may be an active microorganism or an inactive microorganism.

[0109] The fermentation product can be used with extraction or purification, or can be used directly without extraction or purification. If necessary, extraction and purification can be easily achieved using standard extraction and / or purification methods or techniques described in the literature.

[0110] The microorganisms in the inoculant composition may be in active or inactive form and may be vegetative cells, reproductive spores, mycelium, hyphae, conidia or any other form of microbial propagation. The composition may also contain a combination of any of these microbial forms.

[0111] In one embodiment, when a combination of microbial strains is included in the composition, the different strains of microorganisms are grown separately and then mixed together to prepare the composition.

[0112] Advantageously, according to the present invention, the composition may include a culture medium for microbial growth. The composition may be at least 1%, 5%, 10%, 25%, 50%, 75% or 100% growth medium by weight. The amount of biomass in the composition may be any range of, for example, 0% to 100% by weight, including all percentages therebetween.

[0113] Growth of microorganisms and production of biosurfactants according to the invention

[0114] The present invention utilizes methods for the cultivation of microorganisms and the generation of microbial metabolites and / or other byproducts of microbial growth. The present invention further utilizes culture methods suitable for cultivating microorganisms and generating microbial metabolites on a desired scale. These culture processes include, but are not limited to, liquid culture / fermentation, solid state fermentation (SSF) and modifications, hybrids and / or combinations thereof.

[0115] As used herein, "fermentation" refers to the cultivation or growth of cells under controlled conditions. Growth can be aerobic or anaerobic. In a preferred embodiment, the microorganisms are grown using SSF and / or a modified form thereof.

[0116] In one embodiment, the invention provides materials and methods for generating biomass (such as living cell material, etc.), extracellular metabolites (such as small molecules and proteins, etc.), residual nutrients and / or intracellular components (such as enzymes and other proteins, etc.).

[0117] The microbial growth vessel used according to the present invention can be any fermenter or culture reactor for industrial use. In one embodiment, the vessel can have functional controls / sensors or can be connected to functional controls / sensors to measure important factors in the culture process, such as pH, oxygen, pressure, temperature, humidity, microbial density and / or metabolite concentration, etc.

[0118] In another embodiment, the container can also monitor the growth of microorganisms in the container (e.g., measuring the number of cells and the stage of growth). Alternatively, daily samples can be taken from the vessel and counted by techniques known in the art (e.g., dilution plate technique). Dilution plate method is a simple technique for estimating the number of organisms in a sample. This technique can also provide an indicator by which different environments or treatments can be compared.

[0119] In one embodiment, the method includes supplementing the culture with a nitrogen source. The nitrogen source can be, for example, potassium nitrate, ammonium nitrate, ammonium sulfate, ammonium phosphate, ammonia, urea and / or ammonium chloride. These nitrogen sources can be used alone or in combination of two or more.

[0120] The method can provide oxygenation for the growing culture. One embodiment utilizes the slow movement of air to remove low oxygen air and introduce oxygenated air. In the case of submerged fermentation, the oxygenated air can be ambient air supplemented daily by mechanical means including an impeller for mechanically agitating the liquid and an air distributor for supplying gas bubbles to the liquid to dissolve oxygen into the liquid.

[0121] The method may further include supplementing the culture with a carbon source. The carbon source may be a carbohydrate, such as glucose, sucrose, lactose, fructose, trehalose, mannose, mannitol and / or maltose; an organic acid, such as acetic acid, fumaric acid, citric acid, propionic acid, malic acid, malonic acid and / or pyruvic acid; an alcohol, such as ethanol, propanol, butanol, pentanol, hexanol, isobutanol and / or glycerol; a fat and oil, such as soybean oil, rapeseed oil, rice bran oil, olive oil, corn oil, sunflower oil, sesame oil and / or linseed oil. These carbon sources may be used alone or in combination of two or more.

[0122] In one embodiment, the growth factor and trace nutrients of microorganisms are included in the culture medium. When the growth microorganisms cannot generate all the vitamins required for them, this is particularly preferred. Inorganic nutrients may also be included in the culture medium, and the inorganic nutrients include trace elements such as iron, zinc, copper, manganese, molybdenum and / or cobalt. In addition, the source of vitamins, essential amino acids and trace elements may be included in the culture medium in the form of fine powder or coarse powder (for example, corn flour) or in the form of extracts (for example, yeast extract, potato extract, beef extract, soybean extract, banana peel extract, etc.) or in purified form. Amino acids may also be included in the culture medium, for example, amino acids that can be used for the biosynthesis of proteins.

[0123] In one embodiment, the culture medium may also include inorganic salts. Available inorganic salts may be potassium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, magnesium sulfate, magnesium chloride, iron sulfate, iron chloride, manganese sulfate, manganese chloride, zinc sulfate, lead chloride, copper sulfate, calcium chloride, sodium chloride, calcium carbonate and / or sodium carbonate. These inorganic salts may be used alone or in a combination of two or more.

[0124] In some embodiments, the culturing method may further include adding additional acid and / or antimicrobial agents to the culture medium before and / or during the culturing process.Antimicrobial agents or antibiotics are used to protect the culture from contamination.

[0125] Additionally, antifoaming agents may be added to prevent the formation and / or accumulation of foam during submerged cultivation.

[0126] The pH of the mixture should be appropriate for the microorganism of interest. Buffers and pH adjusters (such as carbonates and phosphates) can be used to stabilize the pH near the preferred value. When metal ions are present in high concentrations, it may be necessary to use chelating agents in the culture medium.

[0127] The microorganisms can grow in a planktonic form or in a biofilm form. If there is a biofilm, the vessel can contain a substrate on which the microorganisms can grow in a biofilm state. The system can also have the ability to apply factors that promote and / or improve the growth characteristics of the biofilm (e.g., shear stress).

[0128] The pH of the culture should be appropriate for the microorganism of interest and the soil environment to which the composition will be applied. In some embodiments, the pH is about 2.0 to about 10.0, about 2.0 to about 9.5, about 2.0 to about 9.0, about 2.0 to about 8.5, about 2.0 to about 8.0, about 2.0 to about 7.5, about 2.0 to about 7.0, about 3.0 to about 7.5, about 4.0 to about 7.5, about 5.0 to about 7.5, about 5.5 to about 7.0, about 6.5 to about 7.5, about 3.0 to about 5.5, about 3.25 to about 4.0, or about 3.5. Buffers and pH adjusters (such as carbonates and phosphates) can be used to stabilize the pH near the preferred value.

[0129] In one embodiment, the culturing method is carried out at a temperature of about 5°C to about 100°C, about 15°C to about 60°C, about 20°C to about 50°C, about 20°C to about 45°C, 25°C to about 40°C, 25°C to about 37°C, 25°C to about 35°C, about 30°C to about 35°C, about 24°C to about 28°C, or about 22°C to about 25°C. In one embodiment, the culturing can be carried out continuously at a constant temperature. In another embodiment, the culturing may be affected by temperature changes.

[0130] In one embodiment, the equipment used in the method and the culture process is an aseptic device. The culture equipment (e.g., reactor / vessel) can be separated from a sterilizing device (e.g., autoclave), but connected thereto. The culture equipment can also have a sterilizing device for carrying out in situ disinfection before starting inoculation. Air can be sterilized by methods known in the art. For example, ambient air can pass through at least one filter before being introduced into the vessel. In other embodiments, the culture medium can be pasteurized, or alternatively, not heated at all, but can utilize low water activity and low pH value to control the growth of undesirable bacteria.

[0131] In one embodiment, the present invention further provides a method for producing microbial metabolites by culturing the microbial strain of the present invention under conditions suitable for growth and metabolite production, and optionally purifying the metabolites, wherein the microbial metabolites are biosurfactants, enzymes, proteins, ethanol, lactic acid, β-glucan, peptides, metabolic intermediates, polyunsaturated fatty acids and lipids, etc. The content of the metabolites produced by the method can be at least 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90%, etc.

[0132] The microbial growth byproducts produced by the microorganism of interest can be retained in the microorganism or secreted into the growth medium. The culture medium can include compounds that stabilize the activity of the microbial growth byproducts.

[0133] The biomass content of the fermentation medium may be from 5 g / l to 180 g / l or more, or from 10 g / l to 150 g / l, etc.

[0134] The cell concentration can be, for example, at least 1×10 6 CFU / ml to 1x10 13 CFU / ml, 1x10 7 CFU / ml to 1x10 12 CFU / ml, 1x10 8 CFU / ml to 1x10 11 CFU / ml, or 1x10 9 CFU / ml to 1x10 10 CFU / ml.

[0135] The methods and apparatus for culturing microorganisms and producing microbial byproducts can be performed in a batch process, a quasi-continuous process, or a continuous process.

[0136] In one embodiment, all of the microbial culture composition is removed when the culture is complete (eg, when a desired cell density or density of a particular metabolite is reached). In this batch process, a completely new batch is started when the first batch is harvested.

[0137] In another embodiment, only a portion of the fermentation product is removed at any time. In this embodiment, the biomass with living cells, spores, conidia, hyphae and / or mycelium is retained in the vessel as an inoculant for a new culture batch. The removed composition can be a cell-free culture medium or contain cells, spores or other propagation propagules and / or their combinations. In this way, a quasi-continuous system is created.

[0138] Advantageously, the method does not require complex equipment or high energy consumption. The relevant microorganisms can be cultivated and used on site on a small or large scale, even still mixed with their culture medium.

[0139] Advantageously, microbial-based products can be produced in remote areas. Microbial growth facilities can be operated off-grid by utilizing electricity such as solar, wind and / or hydroelectric power.

[0140] Preparation of microbial-based products

[0141] A microbial-based product of the present invention contains only a fermentation medium containing microorganisms and / or microbial metabolites produced by the microorganisms and / or any residual nutrients. The fermented product can be used directly without extraction or purification. If necessary, extraction and purification can be easily achieved using standard extraction and / or purification methods or techniques described in the literature.

[0142] The microorganisms in the microbial-based products can be in active or inactive form, and can also be somatic cells, reproductive spores, conidia, mycelium, hyphae or any other form of microbial propagation bodies. The microbial-based products can also contain any combination of these forms of microorganisms.

[0143] In one embodiment, different strains of microorganisms are cultured separately and then mixed together to produce a microorganism-based product. The microorganisms may optionally be mixed with the medium in which they were grown and dried prior to mixing.

[0144] In one embodiment, the different strains are not mixed together, but rather applied to the plant and / or its environment as separate microbial-based products.

[0145] The microbial-based products can be used without further stabilization, preservation and storage. Advantageously, the direct use of these microbial-based products maintains high viability of the microorganisms, reduces the possibility of contamination by foreign agents and undesirable microorganisms, and maintains the activity of byproducts of microbial growth.

[0146] After the microbial composition is harvested from the growth container, other components may be added when the harvested product is placed in a vessel or otherwise transported for use. Additives may be, for example, buffers, carriers, other microbial compositions produced in the same facility or in different facilities, viscosity modifiers, preservatives, nutrients for microbial growth, surfactants, emulsifiers, lubricants, solubility modifiers, tracking agents, solvents, bactericides, antibiotics, pH adjusters, chelating agents, stabilizers, UV inhibitors, other microorganisms, and other appropriate additives commonly used in such preparations.

[0147] In one embodiment, buffers including organic acids and amino acids or their salts can be added. Suitable buffers include citrate, gluconate, tartrate, malate, acetate, lactate, oxalate, aspartate, malonate, glucoheptonate, pyruvate, galactate, gluconate, propanol glycolate, glutamate, glycine, lysine, glutamine, methionine, cysteine, arginine and mixtures thereof. Phosphoric acid and phosphorous acid or their salts can also be used. Synthetic buffers are suitable for use, but preferably natural buffers are used, such as organic acids and amino acids or their salts listed above.

[0148] In another embodiment, the pH adjusting agent comprises potassium hydroxide, ammonium hydroxide, potassium carbonate or bicarbonate, hydrochloric acid, nitric acid, sulfuric acid, or a mixture.

[0149] In one embodiment, other components may be included in the formulation, such as an aqueous salt formulation (eg, sodium bicarbonate or carbonate, sodium sulfate, sodium phosphate, sodium hydrogen phosphate).

[0150] Alternatively, the product may be stored before use. The storage time is preferably short. Thus, the storage time may be less than 60 days, 45 days, 30 days, 20 days, 15 days, 10 days, 7 days, 5 days, 3 days, 2 days, 1 day or 12 hours. In a preferred embodiment, if living cells are present in the product, the product is stored at a low temperature of less than 20°C, 15°C, 10°C or 5°C, etc.

[0151] The present invention provides compositions for treating standing water (such as reservoirs and puddles) and soils that are prone to flooding and ponding. More specifically, the present invention provides microbial-derived compositions and their use in preventing and / or mitigating (i.e., discharging) undesirable standing water and / or problems associated with standing water (such as mosquitoes and algae growth, etc.). Advantageously, the compositions and methods of the present invention can be used as environmentally friendly, non-toxic and cost-effective solutions for improving water use efficiency, enhancing soil health and controlling pests.

[0152] In certain embodiments, the present invention provides a water treatment composition comprising a surface active molecule. The water treatment composition can be used for a variety of purposes, including:

[0153] a) wetting agents, which are used to promote the drainage of accumulated water into the soil;

[0154] b) preventive treatments used to enhance the wettability and drainage of soils prior to contact with water, thereby preventing ponding and flooding; and

[0155] c) A pest control agent for controlling mosquitoes, mosquito larvae and / or algae in stagnant water.

[0156] Thus, in certain embodiments, methods are provided for applying water treatment to natural and artificial sources of standing water and / or soil. In some embodiments, the method comprises applying the composition to water accumulated on soil, a water container such as a sink, or a body of water such as a pond. In some embodiments, the method comprises applying the composition to the soil before the soil is contacted with irrigation water or rainfall.

[0157] Advantageously, in addition to reducing standing water and / or controlling pests, in some embodiments, the methods and compositions are effective in increasing water use efficiency and soil flow rates under a variety of conditions, including, for example, compacted soils, drought soils, eroded soils, nutrient-depleted soils, waterlogged soils, fire-damaged soils, hydrophobic soils, and / or soils with plants growing therein.

[0158] In a preferred embodiment, the surfactant molecules of the present invention are microbially derived biosurfactants. The biosurfactants can be applied in purified and / or crude form. The biosurfactants in crude form can include biosurfactants and other cell growth products (including fermentation media generated by culturing microorganisms that generate biosurfactants), etc.

[0159] The biosurfactant according to the method of the present invention can be selected from low molecular weight glycolipids (such as sophorolipids, cellobiose lipids, rhamnolipids, mannoerythritol lipids and trehalose lipids, etc.), lipopeptides (such as surfactin, iturin, fengycin, actin and lichenin, etc.), flavin lipids, phospholipids (such as cardiolipin, etc.), fatty acid esters and high molecular weight polymers (for example, lipoproteins, lipopolysaccharide-protein complexes and polysaccharide-protein-fatty acid complexes), etc.

[0160] In certain specific embodiments, the biosurfactant is a sophorolipid (SLP), such as a lactone SLP, an acidic SLP, an amino acid SLP conjugate, a salt-type SLP, or a derivative of any of these.

[0161] In certain embodiments, the water treatment composition may include additional substances, such as carriers, microbial inoculants, pH adjusters, additional pesticides, herbicides, fertilizers, mineral sources, plant seeds, dyes, stabilizers, emulsifiers, prebiotics, and / or polymers.

[0162] The composition can be applied in combination with an aqueous fluid and applied to water and / or soil. In addition, or as an alternative, the composition can be applied as a dry form of powder or granules, which activate the surface active molecules in the composition when mixed with water.

[0163] In certain embodiments, the water treatment composition, due to its nanoparticle micelle size, is particularly useful for reducing the surface and / or interfacial tension between water and soil, and increasing the porosity of compacted soil. The ultra-small micelle size of the irrigation additive allows water to penetrate into the micro- and nano-pores in hard and compacted soils, thereby opening the pores and thus helping to enhance the mobility of air, nutrients, and water.

[0164] In certain embodiments, the water treatment composition reduces the surface tension of water, thereby promoting the sinking of larval and adult mosquitoes for their control.

[0165] In certain embodiments, the water treatment composition has antimicrobial properties to help control algae growth.

[0166] The compositions and methods of the present invention can be used on farms, pastures, fields, lawns, gardens, golf courses, parks, swamps, ditches, drains, and anywhere that is prone to flooding or standing water, has open containers that collect water, or where standing or stagnant water exists. Advantageously, the compositions and methods of the present invention provide a "green" alternative for pest control while improving water use efficiency and soil health.

Claims

1. A method for preventing and / or mitigating undesirable standing water, the method comprising applying a composition comprising a surface active molecule to standing water or soil, wherein the composition reduces the surface tension of the standing water or water in contact with the soil.

2. The method of claim 1, wherein the surface active molecule is a synthetic surfactant, a microbially derived biosurfactant or a plant derived biosurfactant or a surfactant derived from a natural substrate, and wherein the surface active molecule has a micelle size of less than 25 nm.

3. The method according to claim 1, wherein the surfactant molecule is a biosurfactant selected from glycolipids, lipopeptides, flavin lipids, phospholipids, fatty acid esters, lipoproteins, lipopolysaccharide-protein complexes and polysaccharide-protein-fatty acid complexes.

4. The method of claim 3, wherein the biosurfactant is a sophorolipid.

5. The method of claim 1, wherein the composition is mixed with water and applied to the soil or the standing water through an irrigation system.

6. The method of claim 1, wherein the composition is applied to the soil before or after the soil is contacted with water.

7. The method of claim 1, wherein the composition is applied at a concentration of 10 ppm to 100 ppm relative to the amount of water.

8. The method of claim 1, wherein the composition improves drainage of water into the soil, reduces soil compaction by penetrating pores within the soil, increases porosity of the soil, and reduces surface tension and interfacial tension between the water and the soil.

9. The method of claim 1, further comprising applying a microbial soil treatment composition to the soil, wherein the microbial soil treatment composition comprises one or more beneficial microorganisms capable of colonizing the soil and / or plant roots.

10. The method according to claim 9, wherein the one or more beneficial microorganisms are selected from Trichoderma harzianum, Trichoderma viride, Trichoderma koningii, Trichoderma kweichowii, Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus megaterium, Bacillus polymyxa, Bacillus licheniformis, Bacillus laterosporus, Candida guilliermondii, Meyer yeast of Kalibik, Pichia pastoris, Pichia kudriavzevii, Wickham yeast of anomalous nature and Pasteurella hansdei.

11. The method of claim 9, wherein the beneficial microorganism is Bacillus amyloliquefaciens NRRL B-67928.

12. The method according to claim 9, wherein the beneficial microorganism is Wickhamella anomala NRRL Y-68030.

13. The method of claim 9, wherein the beneficial microorganism is Bacillus subtilis B4 NRRL B-68031.

14. The method of claim 1, wherein the standing water is a puddle, swamp, pond, lake, pool, reservoir, or waterlogged area.

15. The method of claim 1 further used to control pests that breed in the standing water.

16. The method of claim 15, wherein the pest is a mosquito, a midge or an algae.

17. A method of controlling pests, said method comprising applying a composition comprising a surface-active molecule to standing water or soil that is about to come into contact with water, wherein said pests are of a type that reproduces in said standing water or said soil that is about to come into contact with water.

18. The method of claim 17, wherein the standing water is present in a man-made container or has collected on a non-soil surface.

19. The method of claim 17, wherein the standing water is a puddle, swamp, pond, lake, pool, reservoir, or waterlogged area.

20. The method of claim 17, wherein the pest is a mosquito.

21. The method of claim 20, wherein the composition reduces the surface tension of the water, thereby controlling the mosquitoes by killing adult mosquitoes that come to the water surface to lay eggs.

22. The method of claim 20, wherein the composition reduces the surface tension of the water, thereby preventing larvae and pupae from developing into airborne adults.

23. The method of claim 20, wherein the composition reduces the surface tension of the water, thereby preventing mosquito eggs from adhering to surfaces and to each other.

24. The method of claim 17, wherein the surface active molecule is a synthetic surfactant, a microbially derived biosurfactant, or a plant derived biosurfactant, or a surfactant derived from a natural substrate.

25. The method of claim 17, wherein the surface active molecule is a biosurfactant selected from glycolipids, lipopeptides, flavin lipids, phospholipids, fatty acid esters, lipoproteins, lipopolysaccharide-protein complexes and polysaccharide-protein-fatty acid complexes.

26. The method of claim 25, wherein the biosurfactant is a sophorolipid.

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