Composition and method of use of the composition

Description

1 / 55 “COMPOSITIONS AND METHOD OF USE OF THE COMPOSITION” Related Orders

[001] This application claims the benefit of Provisional Patent Application 63 / 493,903, filed on April 3, 2023, the content of which is incorporated in full into this application by reference. Field of Invention

[002] The present invention generally relates to compositions comprising spore-forming microorganisms with humic substances and the use of such compositions. The present invention also relates to methods of using a composition comprising spore-forming microorganisms with humic substances for specialized uses (specific applications), such as increasing nutrient use efficiency and / or nutrient availability for crops. The present invention also relates to applications in which both the microorganisms and the humic substances are delivered to the end-use application by delivery vehicles, including, but not limited to, carriers (hereinafter referred to as vehicles) for mineral fertilizers and / or seeds. Background of the Invention

[003] With global population growth and changing environmental conditions, the discovery of compositions and methods to improve plant growth and productivity has been the focus of the agricultural and agrochemical industry. Given such a dynamic scenario, there is still a need for more effective compositions and methods that demonstrate improvements in plant growth, productivity, and quality.

[004] One method for improving plant growth that can be considered includes the use of biostimulants and biofertilizers. Due to their resilience to a variety of physical and chemical stressors, endospores are a popular inclusion in biostimulant and biofertilizer preparations. Petition 870250089817, dated 02 / 10 / 2025, page 68 / 130 2 / 55 Microbial. Many agricultural formulations based on microbes are supplied in liquid or powder form. However, in practice, the commercial application of spore-forming microbial products presents unique challenges in achieving long shelf life under a variety of storage and use conditions. These challenges include: (a) physical separation, in which the microbial component becomes non-homogeneously distributed in the packaging; (b) stability to environmental contaminants, especially microbial contaminants, including mold, yeast, or others; and (c) stability of the active microbial ingredients themselves under stressful storage and use conditions, both in pre-coating liquid suspensions and in post-coating mineral fertilizer carriers.Although endospores are more resistant to chemical stressors than vegetative bacterial cells, many of them can be labile at concentrated levels of preservatives, given sufficient contact time. Furthermore, once coated in a mineral fertilizer carrier, the endospores must remain viable and adherent to the fertilizer carrier under a variety of environmental conditions to ensure consistent quality and performance in end-use applications, allowing for relevant storage times and conditions in agricultural industry distribution channels.

[005] Various strategies have been employed to achieve the stability of microbial-based agricultural products, but there is still a need for strategies to help spore-forming microbial compositions remain physically and biologically stable during long-term storage and use under a variety of relevant conditions, both as concentrates and after coating in a mineral fertilizer carrier. Brief Description of the Invention

[006] The present invention describes compositions and methods for increasing the survival capacity of microorganisms. Petition 870250089817, dated 02 / 10 / 2025, page 69 / 130 3 / 55 spore formers and improve the biological efficiency of compositions based on spore-forming microorganisms. The present invention provides compositions comprising at least one bacterial additive and at least one humic substance, which provides stability and viability to the bacterial additive while improving the biological efficiency of the product. Microorganisms and humic substances, in each case, can be associated with delivery vehicles, including, but not limited to, mineral fertilizer and / or seed carriers. This disclosure also provides compositions comprising at least one bacterial additive and at least one humic substance coated onto a mineral fertilizer carrier, which will maintain or improve bacterial viability and / or adhesion to the fertilizer carrier under environmental conditions.

[007] Thus, in one aspect, the present disclosure provides compositions comprising at least one bacterial additive and at least one humic substance. In another aspect, the present disclosure provides compositions comprising at least one spore-forming bacterial additive and at least one humic substance. The compositions of the present disclosure provide microbial-based biostimulants or biofertilizers that promote the stability and viability of the bacterial additive, while improving the biological efficiency of the product, and which may give rise to specialized uses (specific applications), such as increased nutritional efficiency and / or nutrient availability for crops.

[008] The present invention provides a composition comprising at least one bacterial additive and at least one humic substance.

[009] In some embodiments of the composition of the invention, at least one bacterial additive comprises an endospore-forming bacterium of the phylum Bacillota. In some embodiments, the hair Petition 870250089817, dated 02 / 10 / 2025, pp. 70 / 130 4 / 55 less one bacterial additive comprises Bacillus spp., Brevibacillus spp., Paenibacillus spp., or Lysinibacillus spp. In some embodiments, at least one bacterial additive comprises a species of Bacillus. In some embodiments, at least one bacterial additive comprises B. amyloliquefaciens (BA), B. subtilis (BS), B. pumilus (BP), B. licheniformis (BL), B. megaterium (BM), and B. thuringiensis (BT), or any combination thereof. In some embodiments, at least one bacterial additive comprises a combination of B. amyloliquefaciens (BA), B. subtilis (BS), B. pumilus (BP), and B. licheniformis (BL). In some embodiments, at least one bacterial additive comprises a combination of B. amyloliquefaciens (BA), B. pumilus (BP), and B. licheniformis (BL). In some embodiments, at least one bacterial additive comprises endospores. In some embodiments, the endospores are suspended in a solution.

[010] In some embodiments of the composition of the invention, said at least one humic substance comprises humic acid, fulvic acid and / or humin. In some embodiments, said at least one humic substance comprises humic acid. In some embodiments, said at least one humic substance comprises fulvic acid. In some embodiments, said at least one humic substance comprises humin. In some embodiments, said at least one humic substance comprises seaweed extract.

[011] In some embodiments of the invention's composition, the composition further comprises a mineral fertilizer. In some embodiments, the mineral fertilizer comprises urea. In some embodiments, the mineral fertilizer comprises phosphorus (P) and potassium (K). Petition 870250089817, dated 02 / 10 / 2025, p. 71 / 130 5 / 55

[012] In some embodiments of the invention's composition, the composition is mixed with a liquid fertilizer or coated onto a dry fertilizer. In some embodiments, the liquid fertilizer or dry fertilizer comprises nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), sulfur (S), magnesium (Mg), iron (Fe), boron (B), chlorine (Cl), manganese (Mn), zinc (Zn), copper (Cu), molybdenum (Mo), nickel (Ni), cobalt (Co), or any combination thereof. In some embodiments, the liquid fertilizer comprises urea, ammonium nitrate, polyphosphate, monoammonium phosphate, diammonium phosphate, potassium chloride, single superphosphate, triple superphosphate, potassium nitrate, potassium sulfate, nitrogen, phosphorus, and potassium mixtures and compounds (“NPK”), or any combination thereof. In some embodiments, the liquid fertilizer comprises a fluid suspension for fertigation or any fertilizer capable of being dissolved.

[013] In some embodiments of the invention's composition, the composition is mixed with a liquid fertilizer or coated onto a dry fertilizer. In some embodiments, the dry fertilizer comprises urea, monoammonium phosphate, diammonium phosphate, potassium chloride, single superphosphate, triple superphosphate, potassium nitrate, potassium sulfate, mixed and compound NPK, or any combination thereof.

[014] In some embodiments of the invention's composition, the composition is coated onto the seeds of a plant.

[015] In some embodiments of the invention's composition, the composition comprises at least one bacterial additive and at least one humic substance present in a ratio in the range of 1:10 to 10:1. In some embodiments, at least one bacterial additive and at least one humic substance are present in a ratio of 1:5. Petition 870250089817, dated 02 / 10 / 2025, page 72 / 130 6 / 55

[016] In some examples of embodiments of the invention's composition, at least one bacterial additive comprises Bacillus spp, Brevibacillus spp, Paenibacillus spp, Lysinibacillus spp or a combination thereof.

[017] In some embodiments of the invention's composition, the agronomic efficiency and / or partial productivity of the composition are increased in relation to an equivalent composition comprising at least one bacterial additive without the humic substance.

[018] In some embodiments of the composition of the invention, the composition improves crop nutrient assimilation, optimizes yield potential, improves root development, improves plant vigor, increases native microbial activity in the soil, improves soil productivity and / or improves soil health compared with plants that were not treated with the composition, or plants that were treated with an equivalent composition comprising at least one bacterial additive without the humic substance.

[019] In some embodiments of the invention's composition, the composition increases the survival capacity of the bacterial additive. In some embodiments, the composition increases: (a) the stability of the calcium contained in the coated dry fertilizer; (b) the stability of the magnesium contained in the coated dry fertilizer; (c) the stability of the manganese contained in the coated dry fertilizer; (d) the stability of the nitrogen contained in the coated dry fertilizer; (e) the stability of the phosphorus contained in the coated dry fertilizer; (f) the absorption of zinc by the crop when applied with the zinc contained in the coated dry fertilizer; and / or (g) the absorption of iron by the crop when applied with the iron contained in the coated dry fertilizer. In some embodiments, the composition improves the absorption of calcium, magnesium, manganese, nitrogen, potassium, iron, and / or zinc by the plant. Petition 870250089817, dated 02 / 10 / 2025, p. 73 / 130 7 / 55 in most soils.

[020] In some embodiments of the invention's composition, the composition increases the survival capacity of a bacterial spore on a hostile surface. In some embodiments, the hostile surface comprises very high pH (basic), very low pH (acidic), very high levels of life-inhibiting chemicals, high levels of incomplete reactions that produced chemicals, urea, single superphosphate and / or triple superphosphate.

[021] In some embodiments of the invention's composition, the composition further comprises a surfactant and / or a wetting agent.

[022] In some embodiments of the composition of the invention, the moisture content of the composition is less than 20%.

[023] In some examples of embodiments of the invention's composition, the composition further comprises one or more micronutrients.

[024] In some embodiments of the composition of the invention, the composition is mixed with a liquid fertilizer or coated with a dry fertilizer.

[025] In some embodiments of the invention's composition, a drying agent is applied after the composition is coated onto a dry fertilizer.

[026] In some embodiments of the invention's composition, the composition further comprises an agent that induces modifications in a microbial phenotype.

[027] In some embodiments of the composition of the invention, at least one bacterial additive comprises B. amyloliquefaciens, B. subtilis, B. pumilus and B. licheniformis in a 1:1:1:1 ratio. In some embodiments, at least one additive Petition 870250089817, dated 02 / 10 / 2025, page 74 / 130 8 / 55 of the bacterial population comprises B. amyloliquefaciens, B. pumilus, and B. licheniformis in a 1:1:1 ratio.

[028] The invention provides a composition comprising: (a) a liquid component comprising the composition of the invention; and (b) a carrier component comprising a mineral fertilizer or seed.

[029] The invention provides a method that utilizes the composition of the invention, wherein the method comprises a specialized use (specific application). In some embodiments, the specialized use comprises increasing the efficiency in the use of nutrients and / or the availability of nutrients to crops. In some embodiments, the nutrient comprises nitrogen, phosphorus, potassium, calcium, magnesium, sulfur, non-essential nutrients and / or micronutrients. In some embodiments, the non-essential nutrients comprise titanium, selenium and / or silicon. Brief Description of the Figures

[030] Figure 1: shows the effect of treating corn with a composition from this publication on corn yield. Corn yield was recorded by weighing the dry grains per experimental plot (experimental bed). Corn grain yield data were recorded in kg per experimental plot. Detailed Description of the Invention

[031] The present invention provides, among other things, compositions and methods for increasing nutrient use efficiency (NUE) and / or nutrient availability to crops, and increasing the survival capacity of spore-forming microbes and improving the biological efficiency of spore-forming microbe-based compositions. The compositions comprise at least one bacterial additive and at least Petition 870250089817, dated 02 / 10 / 2025, page 75 / 130 9 / 55 a humic substance. Compositions and methods are also provided for increasing the survival capacity of spore-forming microbes and improving the biological efficiency of spore-forming microbe-based compositions comprising at least one bacterial additive and at least one humic substance. The compositions and methods described in the present invention provide certain advantages over other compositions and methods, including, but not limited to, increased stability and viability of the bacterial additive, improved biological efficiency of the microbial product, and promotion of desirable microbial phenotypes, which may adventitiously result in specialized uses (specific applications), such as greater nutrient use efficiency and / or greater nutrient availability for crops. Compositions

[032] One aspect of the present invention relates to compositions comprising at least one bacterial additive and at least one humic substance. In some embodiments, the at least one bacterial additive comprises at least one spore-forming bacterial additive. Notably, in some cases, the compositions of the present invention may also provide microbial-based biostimulants or biofertilizers that support the stability and viability of the bacterial additive, while increasing the product's biological efficiency, nutritional efficiency, and / or nutrient availability to crops. Another aspect of the present invention relates to compositions comprising a) a liquid component comprising at least one bacterial additive and at least one humic substance; and b) a carrier component comprising a mineral fertilizer or a seed.Notably, the compositions of the present invention will maintain or improve bacterial viability and / or adhesion to the vehicle under ambient conditions. Petition 870250089817, dated 02 / 10 / 2025, p. 76 / 130 10 / 55

[033] In some embodiments, at least one bacterial additive comprises an endospore-forming bacterium of the phylum Bacillota. In some embodiments, at least one bacterial additive comprises Bacillus spp, Brevibacillus spp, Paenibacillus spp, Lysinibacillus spp and / or another species of Bacillota, wherein spp refers to species within a genus. For example, Bacillus spp refers to one or more species within the genus, such as the genus Bacillus. In some embodiments, at least one bacterial additive is Bacillus spp, Brevibacillus spp, Paenibacillus spp and / or Lysinibacillus spp. In some embodiments, the Bacillus species is B. amyloliquefaciens (BA), B. subtilis (BS), B. pumilus (BP), B. licheniformis (BL), B. megaterium (BM) and B. thuringiensis (BT) or another Bacillus species.

[034] In some embodiments, the bacterial additive comprises one or more endospore-forming bacteria of the phylum Bacillota. Bacillota is a phylum of bacteria that includes the genus Bacillus. In some embodiments, the one or more endospore-forming bacteria of the phylum Bacillota are Bacillus spp, Brevibacillus spp, Paenibacillus spp, Lysinibacillus spp, another species of Bacillota, or any combination thereof. In some embodiments, the one or more endospore-forming bacteria of the phylum Bacillota are of a species of Bacillus, Brevibacillus, Paenibacillus, or Lysinibacillus. In some embodiments, the Bacillus species is B. amyloliquefaciens (BA), B. subtilis (BS), B. pumilus (BP), B. licheniformis (BL), Priestia megaterium (basonym B. megaterium (BM)), B. thuringiensis (BT), another Bacillus species, or any combination of these species. In some embodiments, the Bacillus species is B. amyloliquefaciens (BA), B.subtilis (BS), B. pumilus (BP) and / or B. licheniformis (BL). In some embodiments, the Bacillus species is B. amyloliquefaciens (BA), B. pumilus (BP) and / or B. licheniformis (BL). Petition 870250089817, dated 02 / 10 / 2025, page 77 / 130 11 / 55

[035] Species of the phylum Bacillota, such as Bacillus, Brevibacillus, Paenibacillus, and Lysinibacillus, are known to those skilled in the art. Such species are available in generally recognized deposits. Exemplary information on deposits of bacterial species that can be used in the compositions of the present invention is provided in Table 1 below. Table 1 Deposit Numbers for Specimens of Bacterial Species Species Depository institution Deposit number(s). B. amyloliquefaciens ATCC* 23350, BAA-390, 23842, 23844 licheniformis ATCC 14580, 12713, 9945 * American Type Culture Collection (atcc.org). Endospore Suspensions or Liquid Solutions

[036] In some embodiments, at least one bacterial additive comprises a liquid endospore solution (i.e., in a solution). In some embodiments, the liquid endospore suspension comprises at least one endospore-forming bacterium of the phylum Bacillota. In some embodiments, the one or more endospore-forming bacteria of the phylum Bacillota are Bacillus spp, Brevibacillus spp, Paenibacillus spp, Lysinibacillus spp, another species of Bacillota, or any combination thereof.

[037] In some embodiments, the liquid endospore suspension has a microbial concentration of about 1x10⁶ to about 1x10¹² colony-forming units per mL (cfu / mL), per Petition 870250089817, dated 02 / 10 / 2025, page 78 / 130 12 / 55 example, 1χ106a approximately 1χ1011 cfu / mL, approximately 1χ108a approximately 1χ1012 cfu / mL or approximately 1χ109a approximately 1χ1012 cfu / mL. In some embodiments, the liquid endospore has a microbial concentration of approximately 1χ107a approximately 1χ1011 cfu / mL. In some embodiments, the liquid endospore suspension has a microbial concentration of approximately 1χ108a approximately 1χ1010 cfu / mL. In some embodiments, the liquid endospore suspension has a microbial concentration of approximately 5χ108a approximately 1χ1010 cfu / mL. In some embodiments, the liquid endospore suspension exhibits a microbial concentration of approximately 5χ10⁸ to approximately 5χ10⁹ cfu / mL. Exemplary liquid endospore solutions are described in publication PCT / US2023 / 079253, the content of which is incorporated in full into this application by reference.

[038] In some embodiments, the liquid endospore suspension comprises one or more preservatives, buffering agents, suspending agents, surfactants or a combination thereof. Exemplary additional preservatives, buffering agents, suspending agents and surfactants are described in WO 2021 / 022128, the contents of which are incorporated herein by reference.

[039] One or more preservatives may be selected from a modified isothiazolinone compound, a p-hydroxybenzoic acid ester, a modified quaternary amine, a modified urea, a glycerin derivative, 2-bromo-2-nitro-1,3-propanediol, a natural oil, an organic acid with a molecular weight not exceeding 200 and at least a pKa greater than 4.2, or any combination thereof. In some embodiments, one or more preservatives comprise(s) calcium sulfate.

[040] In some examples of implementation, the suspension of Petition 870250089817, dated 02 / 10 / 2025, page 79 / 130 13 / 55 liquid endospores may include from about 0.01% by weight to 15.0% by weight of one or more preservatives, for example, from about 0.01% by weight to 10.0% by weight, from about 0.01% by weight to 5.0% by weight, from about 0.01% by weight to 3.0% by weight, from about 0.01% by weight to 1.0% by weight, from about 0.1% by weight to 15.0% by weight, from about 0.1% by weight to 10.0% by weight, from about 0.1% by weight to 5.0% by weight, from about 0.1% by weight to 3.0% by weight, from about 0.1% by weight to 1.0% by weight, from about 0.5% by weight to 15.0% by weight, from about 0.5% by weight to 10.0% by weight, about 0.5% by weight to 5.0% by weight, about 0.5% by weight to 3.0% by weight, about 0.5% by weight to 1.0% by weight, about 1.0% by weight to 15.0% by weight, about 1.0% by weight to 10.0% by weight, about 1.0% by weight to 5.0% by weight, or about 1.0% by weight to 3.0% by weight of one or more preservatives.

[041] In some embodiments, the liquid endospore solution may include one suspending agent, two suspending agents, three suspending agents, or more than three suspending agents. The suspending agent is used, among other things, to suspend the microbes in the aqueous composition, thus providing the desired physical stability. In some embodiments, the suspending agent is a polymer, a surfactant, or a combination thereof. In some embodiments, the suspending agent may include xanthan gum, acacia gum, alcohol ethoxylate, or a combination thereof.

[042] In some embodiments, the suspending agent comprises a rheological modifier. Rheological modifiers are substances that alter the rheological properties of a material, i.e., that are added to a formulation to change viscosity. Suitable rheological modifiers may include, among others, commercially available rheological modifiers comprising urea and modified versions of urea, such as RHEOBYK®-410. Petition 870250089817, dated 02 / 10 / 2025, pp. 80 / 130 14 / 55

[043] In some embodiments, the liquid endospore suspension may include from about 0.01% by weight to 15.0% by weight of buffering agent, for example, about 0.01% by weight to 10.0% by weight, about 0.01% by weight to 5.0% by weight, about 0.01% by weight to 3.0% by weight, about 0.01% by weight to 1.0% by weight, about 0.1% by weight to 15.0% by weight, about 0.1% by weight to 10.0% by weight, about 0.1% by weight to 5.0% by weight, about 0.1% by weight to 3.0% by weight, about 0.1% by weight to 1.0% by weight, about 0.5% by weight to 15.0% by weight, about 0.5% by weight 10.0% by weight, about 0.5% by weight to 5.0% by weight, about 0.5% by weight to 3.0% by weight, about 0.5% by weight to 1.0% by weight, about 1.0% by weight to 15.0% by weight, about 1.0% by weight to 10.0% by weight, about 1.0% by weight to 5.0% by weight, or about 1.0% by weight to 3.0% by weight of buffering agent.

[044] The buffering agent may be in sufficient quantity to maintain the pH of the liquid endospore suspension greater than 4.2, for example, greater than 4.5, greater than 5.0, greater than 5.5, greater than 6.0, greater than 6.5 or greater than 7.0. In some embodiments, the buffering agent may be in sufficient quantity to maintain the pH of the composition in the range of about 4.3 to about 12.0, for example, about 4.3 to about 11.0, about 4.3 to about 10.0, about 4.3 to about 9.5, about 4.3 to about 9.0, about 4.3 to about 8.5, about 5.0 to about 12.0, about 5.0 to about 11.0, about 5.0 to about 10.0, about 5.0 to about 9.5, about 5.0 to about 9.0, about 5.0 to about 8.5, about 5.5 to about 12.0, about 5.5 to about 11.0, about 5.5 to about 10.0, about 5.5 to about 9.5, about 5.5 to about 9.0, or about 5.5 to about 8.5.

[045] The pH of a liquid composition or liquid suspension of endospores of the present invention can be selected to maximize the life Petition 870250089817, dated 02 / 10 / 2025, page 81 / 130 15 / 55 useful of spores and / or colonies stored in the composition for extended periods. pH values ​​below about 4 can have a detrimental effect on the stability of certain bacterial spores at elevated temperatures. For example, Bacillus spores can demineralize at low pH, losing their resistance to storage at high temperatures. Therefore, a pH above about 4.2 is desirable to improve the biological stability of Bacillus spores. In some embodiments, the composition including at least one bacterial additive and at least one humic substance increases the survival capacity of a bacterial spore on a hostile surface. In some embodiments, the hostile surface comprises very high pH (basic), very low pH (acidic), very high levels of life-inhibiting chemicals, high levels of incomplete reactions that produced chemicals, urea, single superphosphate, and / or triple superphosphate. Humic Substances

[046] The disclosure provides compositions comprising at least one humic substance. In some embodiments, the at least one humic substance comprises at least one humic acid, fulvic acid, humin, or any combination thereof. In some embodiments, the at least one humic substance comprises at least one humic acid, fulvic acid, and / or humin. In some embodiments, the humic substance is a humic acid, a fulvic acid, or a humin. In some embodiments, the humic substance may include at least one humic substance, at least two humic substances, at least three humic substances, or four or more humic substances. In some embodiments, one or more humic substances comprise seaweed extract. In some embodiments, one or more humic substances are extracted from seaweed extract. Petition 870250089817, dated 02 / 10 / 2025, p. 82 / 130 16 / 55 seaweed. In some embodiments, the humic substance originates from industrial processes. In some embodiments, the humic substance is extracted from leonardite. In some embodiments, the humic substance is extracted by means of redox reactions, dissolution, high-pressure and high-temperature extractions, and / or chemical dissolution with strong acids and bases. In some embodiments, the humic substance may include long-chain and / or short-chain humic acid-derived molecules.

[047] In some embodiments, at least one bacterial additive and at least one humic substance are present in a ratio in the range of about 10:1 to 1:10 in the composition. In some embodiments, the bacterial additive comprises Bacillus spp, Brevibacillus spp, Paenibacillus spp and / or Lysinibacillus spp. In some embodiments, at least one bacterial additive and at least one humic substance are present in a ratio in the range of about 20:1 to 1:20. In some embodiments, at least one bacterial additive and at least one humic substance are present in a ratio in the range of about 15:1 to 1:15. In some embodiments, at least one bacterial additive and at least one humic substance are present in a ratio in the range of about 10:1 to 1:10.In some embodiments, at least one bacterial additive and at least one humic substance are present in a ratio in the range of about 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10. In some embodiments, at least one bacterial additive and at least one humic substance are present in a ratio of about 1:5. In some embodiments, at least one bacterial additive and at least one humic substance are present in a ratio of about 1:1.

[048] In some examples of realization, the composition that Petition 870250089817, dated 02 / 10 / 2025, p. 83 / 130 17 / 55 comprises at least one bacterial additive and at least one humic substance; it further comprises at least one surfactant and / or wetting agent. Examples of wetting agents include, among others, propylene glycol and glycerin. Examples of surfactants include, but are not limited to, a non-ionic surfactant, a primary alkyl alcohol ethoxylate, a secondary alkyl alcohol ethoxylate, a primary alkyl alcohol propoxylate, a secondary alkyl alcohol propoxylate, and / or any combination thereof. In some embodiments, the surfactant has a cloud point of about 30 °C to about 80 °C and a hydrophilic-lipophilic balance of about 5 to about 15. In some embodiments, the surfactant is a C13 branched primary alcohol with an average ethoxylation of 5 to 10, for example, 5, 6, 7, 8, 9, or 10.

[049] In some embodiments, the surfactant includes a fully saturated ethoxylated primary C13 branched alcohol, such as Synperonic® 13 / 7 or Synperonic® 13 / 6. In some embodiments, the surfactant includes an ethoxylated alcohol, such as Ecosurf® EH-6. In some embodiments, the surfactant includes a mixture of 58.0 to 62.0% oligomeric D-glucopyranose decyloctylglucoside and 38.0 to 42.0% water, and contains less than 2% decanol and less than 1.0% octanol, such as Triton® CG110. In some embodiments, the surfactant includes a secondary polyether polyol, such as Tergitol® L-62. In some embodiments, the surfactant includes a secondary alcohol ethoxylate, such as Tergitol® 15-S-12.

[050] In some embodiments, the surfactant includes a nonionic EO / PO alkyl copolymer, such as Tergitol® XDLW.

[051] In some embodiments, the composition comprising at least one bacterial additive and at least one humic substance may include from about 0.1% by weight to 10% by weight of surfactant, for example, from about 0.1% by weight to 5% by weight, 0.1% by weight to 2% by weight, Petition 870250089817, dated 02 / 10 / 2025, page 84 / 130 18 / 55 0.1% by weight to 1% by weight, 0.5% by weight to 1% by weight, or 0.5% by weight to 2% by weight of surfactant.

[052] When used in agronomic applications, the compositions of the present invention can provide a number of desirable characteristics related to reduced deterioration, endospore stability and / or specialized uses, including, but not limited to, increased nutrient use efficiency and / or nutrient availability for crops. Fertilizer Compositions

[053] The disclosure provides fertilizer compositions comprising biostimulants or biofertilizer products based on microorganisms that support the stability and viability of the bacterial additive, while increasing the biological efficiency of the product, nutrient use efficiency and / or nutrient availability to crops.

[054] In some embodiments, the composition comprises a) a liquid component comprising at least one bacterial additive and at least one humic substance; and b) a carrier (support) component. In some embodiments, the carrier component comprises a mineral fertilizer or a seed. In some embodiments, the compositions of the present invention will maintain or improve bacterial viability and / or adherence to the carrier under environmental conditions. In some embodiments, the composition comprising at least one bacterial additive and at least one humic substance is mixed with a liquid fertilizer or coated onto a dry fertilizer.

[055] In some embodiments, the present invention relates to fertilizer compositions comprising a liquid fertilizer in combination with a composition comprising at least one additive. Petition 870250089817, dated 02 / 10 / 2025, page 85 / 130 19 / 55 bacterial and at least one humic substance described in this invention. In other embodiments, the present invention relates to fertilizer compositions comprising a dry fertilizer coated with a composition comprising at least one bacterial additive and at least one humic substance described herein.

[056] In some embodiments, the fertilizer is a mineral fertilizer. In some embodiments, the mineral fertilizer comprises urea, monoammonium phosphate, diammonium phosphate and / or various mixed and compound NPK fertilizers (nitrogen, for example, urea, combined with phosphate and potassium sources). In some embodiments, the fertilizer is an organic fertilizer. In some embodiments, the organic fertilizer comprises fishmeal, poultry guano, cattle manure, compost and / or rock phosphate. Another aspect of the present invention relates to a seed-coated fertilizer composition. In some embodiments, the fertilizer composition comprises a composition comprising at least one bacterial additive and at least one humic substance described in the present invention.In some embodiments, a composition comprising at least one bacterial additive and at least one humic substance is coated onto seeds of any plant. In some embodiments, the compositions described in the present invention can be coated onto seeds of any plant, including, but not limited to, rice, corn, soybeans, onions, sugarcane, tomatoes, barley, lettuce, wheat, potatoes, legumes, or grasses.

[057] In some embodiments, a composition of the present invention may be used as a biostimulant and / or enhanced-efficiency fertilizer component in agronomic applications to promote plant health and vigor and improve crop yield. In some Petition 870250089817, dated 02 / 10 / 2025, page 86 / 130 20 / 55 embodiments, it can be applied to the soil after being coated onto substrates or through irrigation / chemigation processes, or by tank mixing with other agrochemicals, or coated onto seeds. In some embodiments, the composition comprising at least one bacterial additive and at least one humic substance can be coated onto a nutrient fertilizer. For example, the composition comprising at least one bacterial additive and at least one humic substance can be coated onto common fertilizer particles such as urea, monoammonium and diammonium phosphate, ammonium sulfate, compound and blended NPKs, and biosolids. The resulting coated fertilizer particles can then be applied in the field through standard fertilization practices known in the state of the art.Alternatively, the composition comprising at least one bacterial additive and at least one humic substance may be mixed with liquid fertilizers and applied as part of a standard fertilization regime known in the state of the art. The composition comprising at least one bacterial additive and at least one humic substance may be combined with other agrochemicals, such as fungicides, herbicides, or insecticides, in a tank mixture and applied by spraying. The composition comprising at least one bacterial additive and at least one humic substance may also be applied directly to the leaves of a plant by foliar application.

[058] In yet another embodiment, compositions of the present invention comprising at least one bacterial additive and at least one humic substance are added to liquid fertilizers certified for use in organic agriculture (e.g., Phytamin®, PhytaQC®, Tridents Pride®, etc.) and the subsequent mixed liquid is used as part of an organic crop fertility program. In some embodiments, the liquid fertilizer comprises a suspension. Petition 870250089817, dated 02 / 10 / 2025, page 87 / 130 21 / 55 fluid for fertigation in combination with urea, ammonium nitrate, polyphosphate, monoammonium phosphate, diammonium phosphate, potassium chloride, single superphosphate, triple superphosphate, potassium nitrate, potassium sulfate, NPK (mixed and compound) and / or any fertilizer that can be dissolved.

[059] In another embodiment, compositions of the present invention comprising at least one bacterial additive and at least one humic substance are coated onto a dry fertilizer. In some embodiments, the dry fertilizer is an organic fertilizer. In some embodiments, the organic fertilizer includes fishmeal, bird guano, cattle manure, compost, and / or rock phosphate. In some embodiments, the dry fertilizer is a mineral fertilizer. In some embodiments, the mineral fertilizer comprises urea, monoammonium phosphate, diammonium phosphate, and / or various mixed and compound nitrogen, phosphorus, and potassium (“NPK”) compounds. In some embodiments, the dry fertilizer comprises urea, monoammonium phosphate, diammonium phosphate, potassium chloride, single superphosphate, triple superphosphate, potassium nitrate, potassium sulfate, and / or NPK (mixed and compound).

[060] In some embodiments, after the composition comprising at least one bacterial additive and at least one humic substance is coated onto a dry fertilizer, a drying agent is applied (post-coating). In some embodiments, the drying agent is ammonium sulfate, diammonium phosphate, ammonium phosphate, ammonium nitrate, another ammonia-based drying agent, or any combination thereof. In some embodiments, the drying agent is applied to granular urea after coating with a composition comprising at least one bacterial additive and at least one humic substance. Petition 870250089817, dated 02 / 10 / 2025, page 88 / 130 22 / 55

[061] In another embodiment, liquid fertilizer or dry fertilizer comprises nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), sulfur (S), magnesium (Mg), iron (Fe), boron (B), chlorine (Cl), manganese (Mn), zinc (Zn), copper (Cu), molybdenum (Mo), nickel (Ni), cobalt (Co), or any combination thereof.

[062] To improve the efficiency of the coating and the retention of the composition comprising at least one bacterial additive and at least one humic substance on a dry fertilizer with a hydrophobic and / or external environment, a surfactant and / or a wetting agent may be added to the composition. The addition of an appropriate surfactant and / or wetting agent may also aid in steric stabilization, preventing spores from clumping together. In some embodiments, wetting agents comprise propylene glycol and glycerin. In some embodiments, the surfactant comprises a non-ionic surfactant, a primary alkyl alcohol ethoxylate, a secondary alkyl alcohol ethoxylate, a primary alkyl alcohol propoxylate, a secondary alkyl alcohol propoxylate, and / or any combination thereof. Methods of Use of the Compositions of the Invention: Methods of fertilizing crops.

[063] The present invention provides methods for fertilizing crops using any of the compositions described herein. In some embodiments, the method comprises contacting the crop with a composition comprising at least one bacterial additive and at least one humic substance. In some embodiments, the crop is organic. In some embodiments, the crop is not organic. In some embodiments, the crop is grown using conventional agricultural practices (i.e., the uniform management of crop resources, including the use of chemical fertilizers and pesticides). Examples of crops Petition 870250089817, dated 02 / 10 / 2025, page 89 / 130 23 / 55 includes, among others, rice, corn, soybeans, onions, sugarcane, tomatoes, potatoes, barley, wheat, legumes, lettuce, cotton, oats, and grasses.

[064] In some embodiments, the method comprises contacting the crop with the composition comprising at least one bacterial additive and at least one humic substance, at least once during the growing season, for example, 1 to 3 times. In some embodiments, crop contact comprises applying the composition comprising at least one bacterial additive and at least one humic substance to the soil, an irrigation system and / or the leaves of the crop. In some embodiments, the method further comprises mixing the composition comprising at least one bacterial additive and at least one humic substance with a fertilizer before contact with the crop.

[065] When used in agronomic applications, the compositions of the present invention can provide a number of desirable characteristics related to soil health and improved soil quality, such as increased nutrient availability, higher organic matter content, less compaction, and better moisture retention. The composition, which comprises at least one bacterial additive and at least one humic substance, can also provide a number of desirable characteristics related to plant health and vigor, including, among others, improved nutrient absorption, greater tolerance to abiotic stress, greater plant biomass, higher NDVI (Natural Difference Vegetation Index), and higher agricultural productivity.

[066] In some embodiments, the composition including at least one bacterial additive and at least one humic substance may improve crop nutrient assimilation, may optimize yield potential, may increase root development, may improve Petition 870250089817, dated 02 / 10 / 2025, pp. 90 / 130 24 / 55 plant vigor, can increase native microbial activity in the soil, can improve soil productivity and / or can improve soil health. In some embodiments, the composition can improve the assimilation of essential and / or non-essential nutrients by the crop. In some embodiments, the composition improves the assimilation of essential nutrients by the crop. For example, a composition that includes at least one bacterial additive and at least one humic substance improves the assimilation of calcium, magnesium, manganese, nitrogen, phosphorus, zinc, iron, another essential nutrient, or any combination thereof. In some embodiments, a composition that includes at least one bacterial additive and at least one humic substance improves the assimilation of calcium, magnesium, manganese, nitrogen, potassium, phosphorus, zinc, and / or iron by the crop.In some embodiments, the composition improves the uptake of calcium, magnesium, manganese, nitrogen, potassium, iron, and / or zinc by the plant in most soils. In some embodiments, the composition maintains or increases: (a) the stability of the calcium contained in the coated dry fertilizer; (b) the stability of the magnesium contained in the coated dry fertilizer; (c) the stability of the manganese contained in the coated dry fertilizer; (d) the stability of the nitrogen contained in the coated dry fertilizer; (e) the stability of the phosphorus contained in the coated dry fertilizer; (f) the uptake of zinc by the crop when applied with the zinc contained in the coated dry fertilizer; and / or (g) the uptake of iron by the crop when applied with the iron contained in the coated dry fertilizer.

[067] In some embodiments, the composition that includes at least one bacterial additive and at least one humic substance can improve the assimilation of non-essential nutrients by the crop. For example, the composition that includes at least one bacterial additive and at least one humic substance improves the assimilation of titanium, selenium and / or Petition 870250089817, dated 02 / 10 / 2025, pp. 91 / 130 25 / 55 silicon, another non-essential nutrient, or any combination thereof by the crop. In some embodiments, the composition that includes at least one bacterial additive and at least one humic substance improves the assimilation of titanium, selenium, and / or silicon by the crop. In some embodiments, the composition improves the absorption of titanium, selenium, and / or silicon by plants in most soils.

[068] In some embodiments, the method increases nutrient use efficiency and / or the availability of non-essential nutrients to crops. For example, the method increases nutrient use efficiency and / or the availability of non-essential nutrients to crops: titanium, selenium and / or silicon, another non-essential nutrient or any combination thereof. In some embodiments, the method increases nutrient use efficiency and / or the availability of non-essential nutrients to crops: titanium, selenium and / or silicon. In some embodiments, the method increases nutrient use efficiency and / or the availability of non-essential nutrients to crops: titanium, selenium and / or silicon for plant uptake in most soils.

[069] In some embodiments, the method increases nutrient use efficiency and / or nutrient availability for crops, wherein the nutrient includes essential nutrients, non-essential nutrients and / or micronutrients. In some embodiments, the method increases nutrient use efficiency and / or nutrient availability for crops, wherein the nutrient includes nitrogen, phosphorus, potassium, calcium, magnesium, sulfur, non-essential nutrients and / or micronutrients.

[070] In some embodiments, the method increases nutrient use efficiency and / or the availability of essential nutrients for crops. For example, the method increases nutrient use efficiency Petition 870250089817, dated 02 / 10 / 2025, page 92 / 130 26 / 55 nutrients and / or the availability of essential nutrients for the crop: calcium, magnesium, manganese, nitrogen, potassium, phosphorus, zinc, iron, another essential nutrient, or any combination thereof. In some embodiments, the method increases nutrient use efficiency and / or the availability of essential nutrients for the crop: calcium, magnesium, manganese, nitrogen, potassium, phosphorus, zinc, and / or iron. In some embodiments, the method increases nutrient use efficiency and / or the availability of essential nutrients for the crop: calcium, magnesium, manganese, nitrogen, potassium, iron, and / or zinc for plant uptake in most soils.

[071] Another aspect of the present invention relates to the method of using the composition comprising at least one bacterial additive and at least one humic substance, wherein the method comprises a specialized use (specific application). In some embodiments, the specialized use comprises increasing nutrient use efficiency and / or nutrient availability for crops. For example, in some embodiments, the method of using the composition comprising at least one bacterial additive and at least one humic substance increases nutrient use efficiency. In some embodiments, the method of using the composition comprising at least one bacterial additive and at least one humic substance increases nutrient availability for crops. Methods for improving stability

[072] The invention provides methods for inhibiting the proliferation of at least one environmentally spoilage organism using any of the compositions described herein. The compositions of the present invention may be antagonistic to the spoilage organisms while promoting dormancy and endospore stability. Petition 870250089817, dated 02 / 10 / 2025, page 93 / 130 27 / 55

[073] The disclosure also provides methods for inhibiting the proliferation of at least one environmentally spoilage organism in a liquid suspension of endospores, wherein the method comprises the use of at least one osmotic preservative. In some embodiments, the at least one environmentally spoilage organism includes, among others, a bacterial, yeast or fungal contaminant.

[074] In some embodiments, the composition which includes at least one bacterial additive and at least one humic substance can maintain or improve biological stability at about 20 °C to about 40 °C for at least 3 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least one year, at least 1.5 years or at least 2 years. In some embodiments, the composition that includes at least one bacterial additive and at least one humic substance can maintain or improve biological stability in a range of about 20 °C to about 40 °C for about 6 months to about 3 years, for example, from about 6 months to about 2 years, from about 6 months to about 1.5 years, from about 6 months to about 1 year, from about 1 year to about 3 years, from about 1 year to 2 years, or from about 1 year to 1.5 years.In some embodiments, the composition that includes at least one bacterial additive and at least one humic substance can maintain or improve biological stability in a range of about 20 °C to about 40 °C for about 3 months, about 6 months, about 9 months, about one year, about 1.5 years, or about 2 years. For example, the composition that includes at least one bacterial additive and at least one humic substance can maintain or improve biological stability at about 40 °C for about 6 months (i.e., 180 days).

[075] In some embodiments, the composition includes at least one bacterial additive and at least one humic substance. Petition 870250089817, dated 02 / 10 / 2025, pp. 94 / 130 28 / 55 can maintain or improve biological stability at room temperature for at least 3 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least one year, at least 1.5 years, or at least 2 years. In some embodiments, the composition including at least one bacterial additive and at least one humic substance can maintain or improve biological stability at room temperature for about 6 months to about 3 years, for example, about 6 months to about 2 years, about 6 months to about 1.5 years, about 6 months to about 1 year, about 1 year to about 3 years, about 1 year to 2 years, or about 1 year to 1.5 years.For example, a composition including at least one bacterial additive and at least one humic substance can maintain or improve biological stability at room temperature for approximately 3 months, approximately 6 months, approximately 9 months, approximately one year, approximately 1.5 years, approximately 2 years, approximately 2.5 years, or approximately 3 years.

[076] In some embodiments, the composition which includes at least one bacterial additive and at least one humic substance can improve the biological performance of the bacterial additive in a range of about 20 °C to about 40 °C for at least 3 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least one year, at least 1.5 years or at least 2 years. In some embodiments, the composition that includes at least one bacterial additive and at least one humic substance can improve the biological performance of the bacterial additive in a range of about 20 °C to about 40 °C for about 6 months to about 3 years, for example, about 6 months to about 2 years, about 6 months to about 1.5 years, about 6 months to about 1 year, about 1 year to about 3 years, about 1 year to 2 years, or about 1 year to 1.5 years. In some Petition 870250089817, dated 02 / 10 / 2025, pp. 95 / 130 29 / 55 examples of embodiment, the composition that includes at least one bacterial additive and at least one humic substance can improve the biological performance of the bacterial additive in a range of about 20 °C to about 40 °C for about 3 months, about 6 months, about 9 months, about one year, about 1.5 years or about 2 years. For example, the composition that includes at least one bacterial additive and at least one humic substance can improve the biological performance of the bacterial additive at about 40 °C for about 6 months (i.e., 180 days).

[077] In some embodiments, the composition that includes at least one bacterial additive and at least one humic substance can improve the biological performance of the bacterial additive at room temperature for at least 3 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least one year, at least 1.5 years, or at least 2 years. In some embodiments, the composition that includes at least one bacterial additive and at least one humic substance can improve the biological performance of the bacterial additive at room temperature for about 6 months to about 3 years, for example, about 6 months to about 2 years, about 6 months to about 1.5 years, about 6 months to about 1 year, about 1 year to about 3 years, about 1 year to 2 years, or about 1 year to 1.5 years.For example, a composition including at least one bacterial additive and at least one humic substance can improve the biological performance of the bacterial additive at room temperature for approximately 3 months, approximately 6 months, approximately 9 months, approximately one year, approximately 1.5 years, approximately 2 years, approximately 2.5 years, or approximately 3 years.

[078] In some embodiments, the composition that includes at least one bacterial additive and at least one humic substance can improve the biological performance of the bacterial additive. In some Petition 870250089817, dated 02 / 10 / 2025, pp. 96 / 130 In some embodiments, the composition that includes at least one bacterial additive and at least one humic substance increases the survival capacity of the bacterial additive. In some embodiments, the compositions of the present invention may increase the survival capacity of at least one bacterial additive by at least 5% compared to a control composition in which at least one humic substance is not used. In some embodiments, the composition may increase the survival capacity of at least one bacterial additive by at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 20%, at least 30%, at least 40%, or at least 50% compared to a control composition in which at least one humic substance is not used.In some embodiments, the composition can increase the survival capacity of at least one bacterial additive by about 10% to 200%, compared to a control composition in which at least one humic substance is not used. Survival can be measured by plate count assays.

[079] The composition, including at least one bacterial additive and at least one humic substance, can remain biologically and / or physically stable for the periods mentioned above, under a variety of humidity conditions. In some embodiments, the humidity is ambient. In some embodiments, the humidity is from about 10% to about 90%, for example, about 10% to about 80%, about 10% to about 75%, about 10% to about 70%, about 10% to about 65%, about 10% to about 60%, about 20% to about 90%, about 20% to about 80%, about 20% to about 75%, about 20% to about 70%, about 20% to about 65%, about 20% to about 60%, about 30% to about 90%, about 30% to about 80%, about 30% to about 75%, Petition 870250089817, dated 02 / 10 / 2025, p. 97 / 130 31 / 55 approximately 30% to approximately 70%, approximately 30% to approximately 65%, or approximately 30% to approximately 60%. In some embodiments, the humidity is approximately 10%, approximately 15%, approximately 20%, approximately 25%, approximately 30%, approximately 35%, about 40%, about 45%, about 50%, about 55%, about 60%, approximately 65%, or approximately 70%. In some embodiments, the humidity is less than approximately 10%, approximately 15%, approximately 20%, approximately 25%, about 30%, about 35%, about 40%, about 45%, about 50%, approximately 55%, approximately 60%, approximately 65%, or approximately 70%. In some embodiments, the moisture content is less than 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, or 70%. In some embodiments, the composition moisture content is less than 20%.

[080] In some embodiments, the composition maintains or increases: a) the stability of the calcium contained in the coated dry fertilizer; b) the stability of the magnesium contained in the coated dry fertilizer; c) the stability of the manganese contained in the coated dry fertilizer; d) the stability of the nitrogen contained in the coated dry fertilizer; e) the stability of the phosphorus contained in the coated dry fertilizer; or any combination thereof.

[081] Although the present teachings have been described in conjunction with various exemplary realizations (realization examples) and examples, the present teachings should not be understood as limited to the realization examples or examples provided. On the contrary, the present teachings encompass various alternatives, modifications, and equivalents, as will be understood by those skilled in the subject.

[082] Although several examples of embodiments of the invention have been described and illustrated in this document, those skilled in the art will readily foresee a variety of other means and / or structures. Petition 870250089817, dated 02 / 10 / 2025, pp. 98 / 130 32 / 55 for the execution of the function and / or obtaining the results and / or one or more of the advantages described in this document, and each of such variations and / or modifications are considered to be within the scope of the embodiments of the invention described herein. More generally, those skilled in the art will readily perceive that all parameters, dimensions, materials, and configurations described herein are intended to be examples and that the actual parameters, dimensions, materials, and / or configurations will depend on the specific application or applications for which the teachings of the invention are used. Those skilled in the art will recognize many equivalents to the specific inventive embodiments described herein.Therefore, it should be understood that the examples of embodiments described above are presented only by way of example, and that, within the scope of the appended and equivalent claims thereto, examples of embodiments of the invention may be practiced in ways other than those specifically described and claimed. The examples of embodiments of the invention in the present description are directed to each individual feature, article, system, material, kit, and / or method described. Furthermore, any combination of two or more of these elements, systems, articles, materials, kits, and / or methods, provided that such features, systems, articles, materials, kits, and / or methods are not mutually incompatible, shall be included as being within the inventive scope of the present invention. Definitions

[083] All definitions, as defined and used in the present invention, shall be understood as prevailing over dictionary definitions, definitions in documents incorporated by reference, and / or common meanings of the defined terms.

[084] As used in the present invention, “hostile surface” Petition 870250089817, dated 02 / 10 / 2025, pp. 99 / 130 33 / 55 refers to a surface that diminishes the survival capacity of a microorganism, including, but not limited to, bacterial spores. For example, a hostile surface may have an excessively acidic pH or an excessively basic pH. Another example of a hostile surface may include, but not limited to, an excessively smooth particle surface and other retention problems. Furthermore, a hostile surface should be understood as including, but not limited to, a surface hostile to microbial survival and / or retention potential. A non-limiting example of a hostile surface may include, but not limited to, compacted fertilizers and other excessively smooth particles with few hiding places for endospore adsorption.

[085] As used in the present invention, “liquid fertilizer” refers to any form of liquid fertilizer available, any fluid suspension for fertigation, and any fertilizer capable of being dissolved. For example, liquid fertilizer may include, but is not limited to, urea, ammonium nitrate, polyphosphate, monoammonium phosphate, diammonium phosphate, potassium chloride, single superphosphate, triple superphosphate, potassium nitrate, potassium sulfate, and / or NPK (mixed and compound). In addition, liquid fertilizer may include nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), sulfur (S), magnesium (Mg), iron (Fe), boron (B), chlorine (Cl), manganese (Mn), zinc (Zn), copper (Cu), molybdenum (Mo), nickel (Ni), cobalt (Co), or any combination thereof.

[086] As used in the present invention, “dry fertilizer” refers to any form of dry fertilizer available, any fertilizer capable of being mixed and / or compounded to produce a dry mixture. For example, dry fertilizer may include, but is not limited to, urea, monoammonium phosphate, diammonium phosphate, potassium chloride, single superphosphate, triple superphosphate, potassium nitrate, potassium sulfate and / or NPK (mixed and compounded). In addition, dry fertilizer may include nitrogen (N), phosphorus Petition 870250089817, dated 02 / 10 / 2025, pages 100 / 130 34 / 55 (P), potassium (K), calcium (Ca), sulfur (S), magnesium (Mg), iron (Fe), boron (B), chlorine (Cl), manganese (Mn), zinc (Zn), copper (Cu), molybdenum (Mo), nickel (Ni), cobalt (Co) or any combination thereof. The fertilizer may be understood as granular (e.g., granular urea). The fertilizer may be understood as granular (e.g., granular urea).

[087] As used in the present invention, “NPK” refers to nitrogen, phosphorus, and potassium fertilizers, which include the macronutrients nitrogen (N), phosphorus (P), and potassium (K) in varying amounts. The amount of each macronutrient component may vary depending on the plant’s nutritional needs. NPK fertilizers may be compound, in which the components are chemically bonded, or blended, in which the individual nutrient components form a physical mixture.

[088] As used in the present invention, “drying agent” refers to one or more inorganic salts applied to a dry fertilizer. For example, the drying agent may include, among others, ammonium sulfate, diammonium phosphate, ammonium phosphate, ammonium nitrate, another ammonia-based drying agent, or any combination thereof. These compounds may be applied after the application of a composition of the present invention to a fertilizer.

[089] As used in the present invention, “humic substance” refers to one or more humic substances or at least one humic substance. Humic substances refer to a category of compounds that are present in soil and other organic matter. These compounds are not easily decomposed and are known to mediate many chemical reactions and are believed to play a role in reducing soil toxicity. Humic substances may be derived from decomposed plant and animal material, including lignin, tannins, cellulose, and cutins; or they may be produced commercially or industrially from Petition 870250089817, dated 02 / 10 / 2025, pp. 101 / 130 35 / 55 coal, lignite, leonardite, or other organic materials. Humic substances may also be derived from seaweed and / or seaweed extracts (e.g., brown seaweed (kelp)). They are characterized by aromatic, polyaromatic, aliphatic, and carbohydrate units, often with carboxylic, phenolic, and ester functional groups. These substances may include long-chain and / or short-chain humic molecules. See; Trevisan S. et al., Humic substances biological activity at the plant-soil interface: from environmental aspects to molecular factors. Plant Signal Behav. 2010 Jun;5(6):635-43; and Ampong et al., Understanding the Role of Humic Acids on Crop Performance and Soil Health. Front. Agron, 2022; Vol. 4, whose relevant disclosures are incorporated herein by reference for the purposes and subjects mentioned herein.

[090] Furthermore, a humic substance may be understood as including, but not limited to, humic substances extracted by means of industrial processes, including, but not limited to, redox reactions, dissolution, high-pressure and high-temperature extractions, and chemical dissolution with strong acids and bases. A humic substance may include long-chain and / or short-chain humic molecules. In addition, a humic substance may include, but is not limited to, industrial humic substances and non-industrial or commercial humic substances. Industrial humic substances are defined as a preparation of specific molecules from the humic molecular family, sold as single molecules or mixtures of molecules, produced using specific industrial processes.The separation of molecular groups is based on specific chemical processes (condensation, redox, complexation, and many others) that can regroup a supermixture of molecules using industrialized chemical configurations. These molecules, or groups of molecules, can be selected according to their ability to generate specific results. Petition 870250089817, dated 02 / 10 / 2025, pp. 102 / 130 36 / 55 in their end-use application (e.g., absorption of specific nutrients such as P or K) based on the known activity of the molecule or molecular group in question. Industrial humic substances exhibit little or no batch-to-batch variation. Non-industrial or commercial humic substances are defined as a preparation of molecules from the humic molecular family produced by processes targeting this molecular family in general, without considering specific molecular components. The separation of molecular components is based on a set of pH and solubility / precipitation relationships. These molecular mixtures can be selected for their ability to generate generalized results in the end-use application (e.g., growth promotion, nutrient absorption). Non-industrial or commercial humic substances often exhibit significant batch variations.

[091] Examples of humic substances may include humic acid, fulvic acid, humin and / or an industrial humic substance. As used in the present invention, “humic acid” refers to an organic composition, often extracted from humic substances found in soil, which is typically water-soluble at pH 7 or higher and insoluble at pH 2 or lower. Humic acids may comprise carboxylate and / or phenolate groups, which can form complexes with ions in the soil and have been shown to play a role in regulating the bioavailability of metal ions in the soil. See Lovland PJ., Humic Substances: Structures, Models and Functions, The Journal of Agricultural Science. 2002;139(1):113-114., relevant disclosures of which are incorporated herein by reference for the purposes and subject matter hereof.

[092] As used in the present invention, “fulvic acid” refers to an organic compound, often extracted from humus found in soil, which shares many structural similarities with Petition 870250089817, dated 02 / 10 / 2025, pp. 103 / 130 37 / 55 humic acid, but generally has a greater number of carboxylic groups and a lower molecular weight. Fulvic acid's structure allows it to be soluble in water at any pH.

[093] As used in the present invention, “humin” refers to an organic compound, often extracted from humus found in soil, that is not soluble in water at any pH and, when compared to humic acid and fulvic acid, generally has a higher molecular weight and lower levels of functional groups, specifically carboxyl and hydroxyl. See Hayes M et al., Chapter Two - Humin: Its Composition and Importance in Soil Organic Matter, Advan. Agron., 2017; 4:47-138.

[094] As used in this invention, the term “seaweed extract” can be understood as a liquid extract or a dry / powdered extract of one or more varieties of seaweed. Seaweed can be understood as a multicellular macroalgal organism. The variety of seaweed can be red algae (Rhodophyta), green algae (Chlorophyta) or a brown algae (Phaeophyceae, e.g., kelp). The seaweed extract may contain one or more macronutrients, micronutrients, humic substances, including humic acids, phytohormones, soluble alginates and combinations thereof. The seaweed extract can be used as a fertilizer and is commercially available (e.g., Kelpak®). Suitable seaweed extracts are known to those skilled in the art.

[095] As used in the present invention, the term “biologically stable” refers to the microbial titer of a microbial composition that changes no more than 25% (e.g., no more than 20%, no more than 15%, or no more than 10%) after storage at a specific temperature (e.g., at room temperature) for a period of time (e.g., at least 6 months). Petition 870250089817, dated 02 / 10 / 2025, pp. 104 / 130 38 / 55

[096] In addition, the term “organic performance” may include, but is not limited to, product stability and / or the ability of a microbial consortium to retain a concentration of viable and cultivable endospores that meets or exceeds what the product label states.

[097] As used in the present invention, the term nutrient use efficiency (NUE) may also refer to fertilizer use efficiency (FUE). Furthermore, the following formulas may be adopted to express the NUE (unit quantity of nutrient applied) parameters: Agronomic Efficiency (AE) in relation to T1 = (Yield in treatment Tx - Yield in T1) / Unit quantity of nutrient applied; Partial factor productivity (PFP) = Yield / Unit quantity of nutrient applied; where “T1” is a control treatment (e.g., urea) and “Tx” is a single treatment condition. The nutrients applied may be Nitrogen, Phosphorus, and Potassium, among others. As used in the present invention, “yield” can be understood as the amount of agricultural production harvested per unit area of ​​land. The term can also be understood as grain yield (e.g., corn grain yield) or seed ratio (e.g., the multiplication ratio between the number of plants harvested for each seed planted).

[098] As used in the present invention, the term “vegetation index” can be understood as a spectral image transformation of two or more image bands to enhance the contribution of certain green vegetative features, allowing them to be distinguished from other objects in the image. The Vegetation Index can be a simple ratio (SR), which is a ratio between the reflectance recorded in the Infrared bands. Petition 870250089817, dated 02 / 10 / 2025, pp. 105 / 130 39 / 55 Near (NIR) and Red. The Vegetation Index can be a Normalized Difference Vegetation Index (NDVI), which is a method for monitoring green cover in an area calculated by the ratio between the difference and the sum of reflectances in the NIR and Red bands. The Vegetation Index can be a Photochemical Reflectance Index as a measure of light use, which is a measure of efficiency in light use.

[099] As used in the present invention, the term “standard fertilization program” or “standard farmer fertilization program” can be understood as a program comprising treatment with urea.

[100] The indefinite articles “a, an” as used herein in the descriptive report and claims, unless expressly stated otherwise, shall be understood to mean “at least one”. Any ranges quoted herein are inclusive.

[101] The terms “substantially”, “approximately” and “about” used in this descriptive report and in the claims generally mean more or less 10% of the stated value, for example, about 100 would include 90 to 110.

[102] The expression “and / or”, as used in the descriptive report and claims, should be understood as “one or both” of the elements thus associated, i.e., elements that are present conjuncturally in some cases and present disjunctively in other cases. The multiple elements listed with the expression 'and / or' should be understood equivalently, i.e., as encompassing 'one or more' of the elements thus associated. Other elements may optionally be present in addition to the elements specifically identified by the “and / or” clause, whether or not they are related to the elements specifically identified. Thus, as a non-limiting example, a reference to “A and / or B”, when used in conjunction with non-exhaustive (open) language, such as Petition 870250089817, dated 02 / 10 / 2025, pp. 106 / 130 40 / 55 “comprising”, may refer, in one embodiment, to only A (optionally including other elements besides B), in another embodiment, to only B (optionally including elements besides A), in another embodiment, to A and B together (optionally including other elements); etc.

[103] As used herein in the descriptive report and claims, “or” shall be interpreted as having the same meaning as “and / or” as defined above. For example, when separating items in a list, “or” or “and / or” shall be interpreted as being inclusive, i.e., the inclusion of at least one, but also including more than one, from a number or list of elements, and optionally other unlisted items. Only terms that clearly indicate otherwise, such as “only one of” or “exactly one of”, or, when used in the claims, “consisting of”, will refer to the inclusion of exactly one element from a number or list of elements. In general, the term “or” as used herein shall be interpreted as an exclusive alternative indication (e.g., “one or the other, but not both”), when preceded by terms of exclusivity, such as “none of”, “one of”, “only one”, or “exactly one of”."Essentially consisting" when used in claims, has its normal meaning as used in the field of patent law.

[104] As used herein in the descriptive report and claims, the phrase “at least one,” in relation to a list of one or more elements, is to be interpreted as meaning at least one element selected from any one or more elements in the list of elements, but does not necessarily include at least one of each of the elements specifically listed in the list of elements and does not exclude any combinations of the elements in the list of elements. This definition also allows for elements to be present, optionally, in addition to those listed in the list of elements. Petition 870250089817, dated 02 / 10 / 2025, pp. 107 / 130 41 / 55 elements specifically identified in the list of elements to which the phrase “at least one” refers, whether or not they are related to the specifically identified elements. Thus, as a non-limiting example, “at least one of A and B” (or, equivalently, “at least one of A or B”, or equivalently, “at least one of A and / or B”) may refer in one embodiment to at least one A, optionally including more than one A, without the presence of B (optionally including other elements besides B), in another embodiment, to at least one B, optionally including more than one B, without the presence of A (and optionally including other elements besides A), and in yet another embodiment, to at least one A, optionally including more than one A, and at least one B, optionally including more than one B (and optionally including other elements); etc.

[105] In the claims, as well as in the descriptive report above, all transitional phrases, such as “comprising”, “including”, “carrying”, “possessing”, “containing”, “enveloping”, “retaining”, “composed of” and the like, should be understood as being non-exhaustive (open), that is, meaning 'including, but not limited to'. Only the transitional phrases “consisting of” and “consisting essentially of” should be closed or semi-closed transitional phrases, respectively, as set forth in the U.S. Patent Office Manual of Patent Examination Procedures, Section 2111.0.

[106] The claims should not be interpreted as limited to the order or elements described, unless indicated to that effect. It should be understood that various alterations in form and detail may be made by a professional with common knowledge in the field without departing from the spirit and scope of the appended claims. All examples of embodiment that fall within the spirit and scope of the claims to be Petition 870250089817, dated 02 / 10 / 2025, pp. 108 / 130 42 / 55 and equivalents thereof are claimed.

[107] Although methods and materials similar or equivalent to those described herein may be used in the practice or testing of this invention, some illustrative methods and materials are now described. Other features, objectives and advantages of the invention will be evident from the description and claims. In the descriptive report and appended claims, singular forms also include the plural, unless the context clearly indicates otherwise. Unless defined otherwise, the technical and scientific terms used in the present invention have the same meaning as is commonly understood by a person skilled in the art to which this invention pertains. All patents and publications cited in this descriptive report are fully incorporated into this application by reference. Illustrative Achievements

[108] In the present invention, examples of embodiments of compositions and methods for increasing the survival capacity of spore-forming microbes and improving the biological efficiency of spore-forming microbe-based compositions, as described in the present invention, are described. The invention can be understood with reference to the following enumerated embodiments.

[109] Example of embodiment 1. A composition comprising at least one bacterial additive and at least one humic substance, which provides stability and viability to the bacterial additive, while improving the biological efficiency of the product.

[110] Embodiment example 2. The composition, according to embodiment example 1, in which at least one bacterial additive and at least one humic substance can be associated with delivery vehicles, including, but not limited to, mineral fertilizer vehicles. Petition 870250089817, dated 02 / 10 / 2025, pp. 109 / 130 43 / 55

[111] Example embodiment 3. A composition comprising at least one bacterial additive and at least one humic substance coated onto a mineral fertilizer carrier, which will maintain or improve bacterial viability and / or adhesion to the fertilizer carrier under environmental conditions.

[112] Example embodiment 4. A composition comprising at least one spore-forming bacterial additive and at least one humic substance.

[113] Embodiment Example 5. Compositions, according to any of the previous embodiment examples, in which the composition is used as a microbial-based biostimulant or biofertilizer to support the stability and viability of the bacterial additive, while improving the biological efficiency of the product, while also supporting specialized uses (specific applications), such as increased nutrient use efficiency and / or nutrient availability for crops.

[114] Example of embodiment 6. The composition, according to any of the previous examples of embodiment, in which at least one bacterial additive comprises endospore-forming bacteria of the phylum Bacillota.

[115] Example embodiment 7. The composition, according to any of the previous embodiment examples, in which at least one bacterial additive comprises Bacillus spp, Brevibacillus spp, Paenibacillus spp and / or Lysinibacillus spp.

[116] Embodiment example 8. The composition, according to any of the previous embodiment examples, in which at least one humic substance comprises humic acid, fulvic acid and / or humins.

[117] Example of realization 9. The composition, according to any of the previous examples of realization, in which the composition is Petition 870250089817, dated 02 / 10 / 2025, pp. 110 / 130 44 / 55 mixed with a liquid fertilizer or coated over a dry fertilizer.

[118] Example of embodiment 10. The composition, according to any of the previous embodiment examples, in which the composition is coated on seeds of any plant.

[119] Embodiment example 11. The composition, according to embodiment example 9, wherein the liquid fertilizer comprises urea, ammonium nitrate, polyphosphate, monoammonium phosphate, diammonium phosphate, potassium chloride, single superphosphate, triple superphosphate, potassium nitrate, potassium sulfate, NPK (mixed and compound), a fluid suspension for fertigation and / or any fertilizer capable of being dissolved.

[120] Embodiment example 12. The composition, according to embodiment example 9, in which the dry fertilizer comprises urea, monoammonium phosphate, diammonium phosphate, potassium chloride, single superphosphate, triple superphosphate, potassium nitrate, potassium sulfate and / or NPK (mixed and compound).

[121] Embodiment example 13. The composition, according to embodiment example 9, in which the liquid fertilizer or dry fertilizer comprises nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), sulfur (S), magnesium (Mg), iron (Fe), boron (B), chlorine (Cl), manganese (Mn), zinc (Zn), copper (Cu), molybdenum (Mo), nickel (Ni), cobalt (Co) or any combination thereof.

[122] Example embodiment 14. The composition according to any of the previous embodiment examples, wherein the composition comprises at least one bacterial additive and at least one humic substance present in a ratio of 1:10 to 10:1; and wherein the at least one bacterial additive comprises Bacillus spp, Brevibacillus spp, Paenibacillus spp and / or Lysinibacillus spp.

[123] Example of realization 15. The composition, according to Petition 870250089817, dated 02 / 10 / 2025, pp. 111 / 130 45 / 55 any of the previous embodiments, where the composition improves the biological performance of the bacterial additive.

[124] Embodiment Example 16. The composition, in accordance with any of the previous embodiment examples, wherein the composition improves crop nutrient assimilation, optimizes yield potential, improves root development, improves plant vigor, improves native microbial activity in the soil, improves soil productivity and / or improves soil health.

[125] Embodiment example 17. The composition, according to any of the previous embodiment examples, in which the composition increases the survival capacity of the bacterial additive.

[126] Embodiment Example 18. The composition, according to any of the previous embodiment examples, in which the composition increases: a) the stability of the calcium contained in the coated dry fertilizer; b) the stability of the magnesium contained in the coated dry fertilizer; c) the stability of the manganese contained in the coated dry fertilizer; d) the stability of the nitrogen contained in the coated dry fertilizer; e) the stability of the phosphorus contained in the coated dry fertilizer; f) the absorption of zinc by the crop when applied with the zinc contained in the coated dry fertilizer; and / or g) the absorption of iron by the crop when applied with the iron contained in the coated dry fertilizer.

[127] Embodiment example 19. The composition, according to any of the previous embodiment examples, in which the composition improves the uptake of calcium, magnesium, manganese, nitrogen, potassium, iron and / or zinc by the plant in most soils.

[128] Embodiment example 20. The composition, according to any of the previous embodiment examples, in which the composition increases the survivability of a bacterial spore on a hostile surface. Petition 870250089817, dated 02 / 10 / 2025, pp. 112 / 130 46 / 55

[129] Embodiment example 21. The composition, according to embodiment example 20, in which the hostile surface comprises very high pH (basic), very low pH (acidic), very high levels of life-inhibiting chemicals, high levels of incomplete reactions that produced chemicals, urea, single superphosphate and / or triple superphosphate.

[130] Embodiment example 22. The composition, according to any of the previous embodiment examples, wherein the composition further comprises a surfactant and / or a wetting agent.

[131] Embodiment example 23. The composition, according to any of the previous embodiment examples, in which the composition moisture is less than 20%.

[132] Example embodiment 24. A method of using a composition comprising at least one bacterial additive and at least one humic substance, wherein the method comprises a specialized use (specific application).

[133] Embodiment example 25. The method, according to embodiment example 24, in which specialized use (specific application) comprises greater efficiency in nutrient use and / or nutrient availability for crops.

[134] Embodiment example 26. The method, according to embodiment example 24, in which the nutrient comprises nitrogen, phosphorus, potassium, calcium, magnesium, sulfur, non-essential nutrients and / or micronutrients.

[135] Embodiment example 27. The method, according to embodiment example 26, in which non-essential nutrients comprise titanium, selenium and / or silicon. Examples

[136] Disclosure is further illustrated by the following examples, which should not be interpreted as limiting the scope or spirit. Petition 870250089817, dated 02 / 10 / 2025, pp. 113 / 130 47 / 55 of this invention to the specific procedures described herein. It should be understood that the examples are provided to illustrate certain embodiments and are not intended to limit the scope of the invention. It should also be understood that various other embodiments, modifications and equivalents that may be suggested by skilled individuals in the field may be used without departing from the spirit of the present invention and / or the scope of the appended claims. Example 1 Evaluation of the Effect of the Compositions of the Present Invention on Corn Cultivation

[137] Several exemplary compositions and methods for their use were devised. The details of the treatments with the tested compositions are summarized in Table 2. The exemplary composition BW F1 comprises Bacillus amyloliquefaciens (1.25E+09 CFU / mL, or colony-forming units per milliliter), Bacillus subtilis (1.25E+09 CFU / mL), Bacillus pumilus (1.25E+09 CFU / mL) and Bacillus licheniformis (1.25E+09 CFU / mL), totaling a content of 5.00E+09 CFU / mL of Bacillus. The exemplary composition BW F2 comprises Bacillus amyloliquefaciens (1.25E+09 CFU / mL), Bacillus pumilus (1.25E+09 CFU / mL) and Bacillus licheniformis (1.25E+09 CFU / mL), totaling a content of 3.75E+09 CFU / mL of Bacillus. The compositions of the present invention can be produced by any of the standard processes known in the art. Table 2 Composition and Treatment Details Treatment No. Treatment Code 1 Dose, kg / ha* Application Time A Application Time B Application Time C T1 Urea 435 (200 kg of N 100% N) 13 days after sowing (BbCH 32, 2 detectable nodes) 21 days after sowing (BBCH 33, 3 detectable nodes) 40 days after sowing (BBCH 39, 9 or more detectable nodes) Petition 870250089817, dated 02 / 10 / 2025, pp. 114 / 130 48 / 55 Treatment No. Treatment Code Dose, kg / ha* Application Time A Application Time B Application Time C T2 Urea+S-Humic 435 (200 kg of N 100% N) 13 days after sowing (BbCH 32, 2 detectable nodes) 21 days after sowing (BBCH 33, 3 detectable nodes) 40 days after sowing (BBCH 39, 9 or more detectable nodes) T3 Urea+BW F1 (435 (200 kg of N 100% N) 13 days after sowing (BbCH 32, 2 detectable nodes) 21 days after sowing (BBCH 33, 3 detectable nodes) 40 days after sowing (BBCH 39, 9 or more detectable nodes) T4 Urea+S-Humic+BW F1 435 (200 kg of N) 100% N) 13 days after sowing (BbCH 32, 2 detectable nodes) 21 days after sowing (BBCH 33, 3 detectable nodes) 40 days after sowing (BBCH 39, 9 or more detectable nodes) T5 Urea+BW F2 435 (200 kg of N 100% N) 13 days after sowing (BbCH 32, 2 detectable nodes) 21 days after sowing (BBCH 33, 3 detectable nodes) 40 days after sowing (BBCH 39,9 or more detectable nodes) T6 Urea+S-Humic+BW F2 435 (200 kg of N 100% N) 13 days after sowing (BbCH 32, 2 detectable nodes) 21 days after sowing (BBCH 33, 3 detectable nodes) 40 days after sowing (BBCH 39, 9 or more detectable nodes) T7 Urea+S-Humic 326 (150 kg of N 75% N) 13 days after sowing (BbCH 32, 2 detectable nodes) 21 days after sowing (BBCH 33, 3 detectable nodes) 40 days after sowing (BBCH 39, 9 or more detectable nodes) T8 Urea+BW F1 326 (150 kg of N 75% N) 13 days after sowing (BbCH 32, 2 detectable nodes) 21 days after sowing (BBCH 33, 3 detectable nodes) 40 days after sowing (BBCH 39, 9 or more detectable nodes) T9 Urea+S-Humic+BW F1 326 (150 kg N 75% N) 13 days after sowing (BbCH 32, 2 detectable nodes) 21 days after sowing (BBCH 33, 3 detectable nodes) 40 days after sowing (BBCH 39, 9 or more detectable nodes) T10 Urea+BW F2 326 (150 kg N 75% N) 13 days after sowing (BbCH 32,2 detectable nodes) 21 days after sowing (BBCH 33, 3 detectable nodes) 40 days after sowing (BBCH 39, 9 or more detectable nodes) T11 Urea+S-Humic+BW F2 326 (150 kg of N 75% of N) 13 days after sowing (BbCH 32, 2 detectable nodes) 21 days after sowing (BBCH 33, 3 detectable nodes) 40 days after sowing (BBCH 39, 9 or more detectable nodes) * kg / ha refers to kilograms per hectare; ** N = nitrogen.

[138] In an experiment, the effectiveness of combinations of S-humic (humic substance) and / or BiOWiSH® Crop Liquid containing biofertilizers (fertilizers containing Bacillus, as described in Petition 870250089817, dated 02 / 10 / 2025, pages 115 / 130 49 / 55 of the present invention) in combination with urea in a maize crop (see Table 2), focusing on evaluating the effect on maize growth and yield under field conditions. This experiment was conducted in a typical field with an alluvial soil type with a known fertilization level (also called fertility level) in order to compare a standard farmer fertilizer program using urea with treatment conditions that included BiOWiSH® Technologies urea formulations coated with BiOWiSH® Crop Liquid and / or S-Humic biofertilizers. The tested rates were 100% and 75% N via Urea, as defined below, with a uniform application of phosphorus (P) and potassium (K) fertilizer in all treatment conditions. Pest and disease control and irrigation management practices were implemented as needed following standard processes known in the state of the art.

[139] The eleven treatments used in this experiment were: (T1) Untreated control (CNT) FERTILIZATION LEVEL - 100% Urea (sometimes referred to as 100% N (via urea), i.e., with urea applied at 200 kg of nitrogen (N) per hectare); (T2) FERTILIZATION LEVEL - 100% Urea + S-HUMIC, (T3) FERTILIZATION LEVEL - 100% Urea + BW F1, (T4) FERTILIZATION LEVEL - 100% Urea + S-HUMIC + BW F1, (T5) FERTILIZATION LEVEL - 100% Urea + BW F2, (T6) FERTILIZATION LEVEL - 100% Urea + S-HUMIC + BW F2, (T7) FERTILIZATION LEVEL - 75% Urea (sometimes referred to as 75% N (via urea), i.e., with urea applied at 150 kg of N per hectare) + S-HUMIC, (T8) FERTILIZATION LEVEL - 75% Urea + BW F1, (T9) FERTILIZATION LEVEL - 75% Urea + S-HUMIC + BW F1, (T10) FERTILIZATION LEVEL - 75% Urea + BW F2, and Petition 870250089817, dated 02 / 10 / 2025, pp. 116 / 130 50 / 55 (T11) FERTILIZATION LEVEL - 75% Urea, S-UMICA + BW F2.

[140] Three applications were made at 13, 21 and 40 days after sowing, when the indicated number of nodes on the plants was already evident (see Table 2).

[141] Soil samples were collected before sowing and analyzed for several parameters to assess nutritional status. The vegetation index (NDVI) was recorded at the 4-node and 6-node stages using a Trimble Green Seeker NDVI meter (Model PSAI10R-050Q) and measuring the index on 10 randomly selected plants in each experimental plot. Plant counts were observed at the 4- and 6-node stages and at harvest. In addition, soil samples up to 12 inches and 12 to 24 inches deep were collected at the heading stage for analysis of soil pH, organic matter content, and nitrate nitrogen content. Plant materials (opposite leaf sample and below the ear) were collected at the heading stage and analyzed for nutrient content such as N, P, K, calcium (Ca), magnesium (Mg), sulfur (S), sodium (Na), copper (Cu), zinc (Zn), manganese (Mn), iron (Fe), and boron (B).Soil and corn kernel samples were also analyzed at harvest time for various parameters. Corn kernel yield was recorded at harvest.

[142] The effect of the treatments on corn yield was evaluated. Corn yield ranged from 6.85 to 7.97 kg / experimental plot (Table 3 and Figure 1). The differences in yield observed between treatments compared to the yield of the untreated control clearly demonstrate the effect of the formulation that included S-Humic with urea coated with BiOWiSH® Crop Liquid containing biofertilizers on corn yield. The treatments FERTILIZATION LEVEL - 100% Urea + S-HUMIC + BW F1 (T4), FERTILIZATION LEVEL - 100% Urea + BW F2 (T5) and FERTILIZATION LEVEL - 100% Urea + S-HUMIC + BW F2 (T6) Petition 870250089817, dated 02 / 10 / 2025, pages 117 / 130 51 / 55 showed relatively much higher yields than the other treatments. The yield performance of all BiOWiSH® product treatments was significantly higher than the control with urea alone. Among the BiOWiSH® treatments, BW F1 in combination with FERTILIZATION LEVEL - 100% urea + S-HUMIC (T4), BW F2 + FERTILIZATION LEVEL - 100% urea (T5), and BW F2 with FERTILIZATION LEVEL - 100% urea + S-HUMIC (T6) are suitable for improving corn productivity. Furthermore, the 100% nitrogen application level was higher compared to the 75% nitrogen application level. However, even with a 75% N level, the use of BiOWiSH® products containing BW F1 and BW F2 resulted in significantly higher corn yields than products without BW F1 and BW F2. Table 3 Effect of Treatments on Corn Productivity Treatment No. Treatment Treatment Code Dose (MU) Yield. Grain Yield Increase compared to the control (%) kg / plot t / ha T1 FERTILIZATION LEVEL 100% Urea Urea 100% N 6.85 f 3.80 f 0.0 T2 FERTILIZATION LEVEL 100% Urea S-Humic Urea+Humic 100% N + Humic 7.23 of 4.02 of 5.5 T3 FERTILIZATION LEVEL - 100% Urea + BW F1 Urea+BW F1 100% N + 0.2% w / w 7.67 bc 4.26 bc 11.9 T4 FERTILIZATION LEVEL 100% Urea S-Humic + BW F1 Urea+Humic+BW F1 100% N + Humic + 0.2% w / w 7.97 to 4.43 to 16.4 T5 FERTILIZATION LEVEL - 100% Urea + BW F2 Urea + BW F2 100% N + 0.2% w / w 7.75 ab 4.30 ab 13.1 T6 FERTILIZATION LEVEL - Urea + Shumic Acid + BW 100% N + S - 7.74 ab 4.30 ab 13.1 Petition 870250089817, dated 02 / 10 / 2025, pp. 118 / 130 52 / 55 Treatment No. Treatment Treatment Code Dose (MU) Yield. Grain Yield Increase compared to the control (%) kg / plot t / ha 100% Urea S-Humic + BW F2 F2 Humic + 0.2% w / w T7 FERTILIZATION LEVEL 75% Urea S-Humic Urea+Humic 75% N + S-Humic 7.10 ef 3.95 ef 3.7 T8 FERTILIZATION LEVEL 75% Urea + BW F1 Urea+BW F1 75% N + 0.2% w / w 7.43 cd 4.13 cd 8.5 T9 FERTILIZATION LEVEL 75% Urea S-Humic + BW F1 Urea+Humic+BW F1 75% N + S-Humic + 0.2% w / w 7.61 bc 4.23 bc 11.2 T10 FERTILIZATION LEVEL 75% Urea + BW F2 Urea + BW F2 75% N + 0.2% w / w 7.44 cd 4.13 cd 8.6 T11 FERTILIZATION LEVEL 75% Urea S-Humic + BW F2 Urea + S-Humic + BW F2 75% N + S-Humic + 0.2% w / w 7.43 cd 4.13 cd 8.4 DC (P=0.05) 0.25 0.14 3.6 DC: Critical difference. Means followed by different letters are significantly different from each other.

[143] The effect of different treatments on nutrient use efficiency (NUE, also known as fertilizer use efficiency, or FUE) was also evaluated. Based on the yield data generated in the experiment, the NUE parameters were calculated to understand the effect of the various treatments on the NUE of N, P, and K. The results obtained are presented in Table 4. The agronomic efficiency (AE) of N supplied through different test items revealed that the treatments containing BW F1 and BW F2 were generally superior at both the 100% rate. Petition 870250089817, dated 02 / 10 / 2025, pages 119 / 130 53 / 55 as well as at the 75% N rate. Partial factor productivity (PFP) revealed that N applied at the 75% N rate was superior to the 100% N rate. Considering that uniform rates of P and K were applied to all treatments, the trend of the agronomic efficiency (AE) and partial factor productivity (PFP) parameters of P and K presented in Table 4 is similar to the yield trend observed in Table 3. Table 4 Effect of Treatments on Agronomic Efficiency and Partial Factor Productivity O c Φ E <ü * do milho । a) Yield) T1 (kg / ha) Nitrogen Phosphorus Potassium Treatment No. Yield (kg / ha) Difference in relation to applied N kg / ha *ea **PPF Applied P2O5 (kg / ha) *ea **ppf Applied K2O (kg / ha) *ea **PPF T1 3805 0 200 0.00 19.0 70 0.0 54.4 70 0.0 54.4 T2 4016 211 200 1.06 20.1 70 3.0 57.4 70 3.0 57.4 T3 4259 454 200 2.27 21.3 70 6.5 60.8 70 6.5 60.8 T4 4430 626 200 3.13 22.2 70 8.9 63.3 70 8.9 63.3 T5 4305 500 200 2.50 21.5 70 7.1 61.5 70 7.1 61.5 T6 4301 497 200 2.48 21.5 70 7.1 61.4 T7 3947 143 150 0.95 26.3 425 150 2.83 28.2 70 6.1 60.4 70 6.1 60.4 T10 4133 329 150 2.19 27.6 70 4.7 59.0 70 4.7 59.0 T11 4126 321 150 2.14 27.5 70 4.6 58.9 70 4.6 58.9 * EA = Agronomic Efficiency in relation to T1 = (Yield in treatment Tx - Yield in T1) / nutrient applied; **PPF - Partial Productivity of the Factor = Yield / Nutrient Applied.**

[144] The results of the soil analysis indicated that the soil was Petition 870250089817, dated 02 / 10 / 2025, pages 120 / 130 The soil was 54 / 55 alkaline and saline, and the availability of primary and secondary nutrients was considered adequate. Soil analysis showed that the application of the different treatments did not demonstrate substantial differences in the levels of the different parameters. Foliar analysis results indicated that, among the main nutrients, N and K levels were relatively low in the leaves, while Ca, Mg, and S levels were more than adequate. Despite the low availability of micronutrients in the soil, their levels in the leaves indicated that the crop was able to absorb these nutrients effectively from the soil.

[145] The quality parameters of the grains analyzed, such as moisture, crude protein, ash, crude fiber and crude fat, indicated that the different treatments did not present a substantial difference in the levels of the different grain quality parameters.

[146] The application of FERTILIZATION LEVEL - 100% Urea SHUMIC + BW F1, FERTILIZATION LEVEL - 100% Urea S-HUMIC + BW F2 and FERTILIZATION LEVEL - 100% Urea + BW F2 resulted in higher yields compared to the other treatments, including the control. Furthermore, the application of BiOWiSH® products containing urea, BW F1 and BW F2, at a 100% N rate was superior to that at a 75% N rate. The agronomic efficiency of N supplied through different test compositions revealed that the use of BiOWiSH® products containing BW F1 and BW F2 was generally superior to both a 100% N rate and a 75% N rate. Partial factor productivity revealed that N applied at a 75% N rate was superior to the 100% N rate. This appears to reflect the law of diminishing returns after a certain level of N application, under the soil conditions in which the crop was grown.

[147] Without intending to limit itself to a specific theory, the results demonstrate that the compositions of the present invention resulted in improved corn crop yield. The improvement in yield of Petition 870250089817, dated 02 / 10 / 2025, pp. 121 / 130 55 / 55 corn cultivation can be the pinnacle of efficient use of N applied through these compositions, as well as other nutrients applied / available in the soil. Furthermore, without intending to limit itself to a specific theory, the results related to corn grain yield demonstrated the significant positive effect of different compositions of the present invention (BiOWiSH® Technologies Formulation + S-Humic with urea coated with BiOWiSH® Crop Liquid containing biofertilizers). Additionally, the application of FERTILIZATION LEVEL - 100% urea + S-HUMIC + BW F1 registered significantly maximum grain yield, followed by the application of FERTILIZATION LEVEL - 100% urea + BW F2 and FERTILIZATION LEVEL 100% Urea + S-HUMIC + BW F2. Without intending to limit itself to theory, the results demonstrate that BiOWiSH® urea products containing BW F1 and BW F2 are suitable for improving corn productivity. Equivalents

[148] Although the present invention has been described in conjunction with the specific embodiments set forth above, many alternatives, modifications and other variations thereof will be apparent to those skilled in the art. All such alternatives, modifications and variations are intended to fit within the spirit and scope of the present invention. Petition 870250089817, dated 02 / 10 / 2025, pages 122 / 130

Claims

1 / 6 Claims 1. COMPOSITION, characterized by comprising at least one bacterial additive and at least one humic substance.

2. COMPOSITION, according to claim 1, characterized in that at least one bacterial additive comprises an endospore-forming bacterium of the phylum Bacillota.

3. COMPOSITION, according to claim 1, characterized by at least one bacterial additive comprising Bacillus spp, Brevibacillus spp, Paenibacillus spp or Lysinibacillus spp.

4. COMPOSITION, according to claim 1, characterized in that at least one bacterial additive comprises a species of the genus Bacillus.

5. COMPOSITION, according to claim 1, characterized by at least one bacterial additive comprising B. amyloliquefaciens (BA), B. subtilis (BS), B. pumilus (BP), B. licheniformis (BL), B. megaterium (BM) and B. thuringiensis (BT), or any combination of these species.

6. COMPOSITION, according to claim 1, characterized in that at least one bacterial additive comprises a combination of B. amyloliquefaciens (BA), B. subtilis (BS), B. pumilus (BP) and B. licheniformis (BL).

7. COMPOSITION, according to claim 1, characterized by at least one bacterial additive comprising a combination of B. amyloliquefaciens (BA), B. pumilus (BP) and B. licheniformis (BL).

8. COMPOSITION, according to any one of claims 1 to 7, characterized by at least one bacterial additive comprising endospores, wherein, optionally, the endospores are suspended in a solution.

9. COMPOSITION, according to any of the preceding claims, characterized by at least one humic substance comprising humic acid, fulvic acid and / or humin.

10. COMPOSITION, according to any of the preceding claims, characterized in that the composition further comprises a mineral fertilizer.

11. COMPOSITION, according to claim 10, characterized in that the mineral fertilizer comprises urea.

12. COMPOSITION, according to any one of claims 10 or 11, characterized by the mineral fertilizer comprising phosphorus (P) and potassium (K).

13. COMPOSITION, according to any one of claims 1 to 12, characterized in that the composition is mixed with a liquid fertilizer or coated with a dry fertilizer.

14. COMPOSITION, according to claim 13, characterized in the liquid fertilizer or dry fertilizer comprising nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), sulfur (S), magnesium (Mg), iron (Fe), boron (B), chlorine (Cl), manganese (Mn), zinc (Zn), copper (Cu), molybdenum (Mo), nickel (Ni), cobalt (Co) or any combination thereof.

15. COMPOSITION, according to claim 13, characterized by the liquid fertilizer comprising urea, ammonium nitrate, polyphosphate, monoammonium phosphate, diammonium phosphate, potassium chloride, single superphosphate, triple superphosphate, potassium nitrate, potassium sulfate, nitrogen, phosphorus and potassium mixed and compounded (“NPK”), or any combination thereof.

16. COMPOSITION, according to claim 15, Petition 870250089817, dated 10 / 02 / 2025, pp. 124 / 130 3 / 6, characterized by the liquid fertilizer comprising a fluid suspension for fertigation, or any fertilizer capable of being dissolved.

17. COMPOSITION, according to claim 13, characterized by the dry fertilizer comprising urea, monoammonium phosphate, diammonium phosphate, potassium chloride, single superphosphate, triple superphosphate, potassium nitrate, potassium sulfate, mixed and compound NPK, or any combination thereof.

18. COMPOSITION, according to any of the preceding claims, characterized in that the composition is coated onto the seeds of a plant.

19. COMPOSITION, according to any of the preceding claims, characterized in that the composition comprises at least one bacterial additive and at least one humic substance present in a ratio in the range of 1:10 to 10:1, wherein, optionally, at least one bacterial additive and at least one humic substance are present in a ratio of 1:

5.

20. COMPOSITION, according to claim 19, characterized by at least one bacterial additive comprising Bacillus spp, Brevibacillus spp, Paenibacillus spp, Lysinibacillus spp or a combination of one or more of these genera.

21. COMPOSITION, according to any of the preceding claims, characterized in that the agronomic efficiency and / or partial productivity of the composition factor are increased in relation to an equivalent composition comprising at least one bacterial additive without the humic substance.

22. COMPOSITION, according to any of the preceding claims, characterized by the composition improving nutrient assimilation by the crop, optimizing yield potential, Petition 870250089817, dated 02 / 10 / 2025, page 125 / 130 4 / 6 increasing root development, improving plant vigor, increasing native microbial activity in the soil, improving soil productivity and / or improving soil health compared to plants that were not treated with the composition, or plants that were treated with an equivalent composition comprising at least one bacterial additive without the humic substance.

23. COMPOSITION, according to any of the preceding claims, characterized by the composition increasing the survival capacity of the bacterial additive.

24. COMPOSITION, according to any one of claims 13 to 23, characterized by the composition increasing: a) the stability of calcium contained in the coated dry fertilizer; b) the stability of magnesium contained in the coated dry fertilizer; c) the stability of manganese contained in the coated dry fertilizer; d) the stability of nitrogen contained in the coated dry fertilizer; e) the stability of phosphorus contained in the coated dry fertilizer; f) the absorption of zinc by the crop when applied with zinc contained in the coated dry fertilizer; and / or g) the absorption of iron by the crop when applied with iron contained in the coated dry fertilizer.

25. COMPOSITION, according to any of the preceding claims, characterized by the composition improving the absorption of calcium, magnesium, manganese, nitrogen, potassium, iron and / or zinc by the plant in most soils.

26. COMPOSITION, according to any of the previous claims, characterized by the composition increasing the survival capacity of a bacterial spore on a hostile surface.

27. COMPOSITION, according to claim 26, characterized by the hostile surface comprising a very high pH (basic), a very low pH (acidic), a very high level of life-inhibiting chemicals, high levels of incomplete reactions that produce chemicals, urea, single superphosphate and / or triple superphosphate.

28. COMPOSITION, according to any of the preceding claims, characterized in that the composition further comprises a surfactant and / or a wetting agent.

29. COMPOSITION, according to any of the preceding claims, characterized in that the moisture content of the composition is less than 20%.

30. COMPOSITION, according to any of the preceding claims, characterized in that the composition further comprises one or more micronutrients.

31. COMPOSITION, according to claim 30, characterized in that the composition is mixed with a liquid fertilizer or coated onto a dry fertilizer.

32. COMPOSITION, according to claim 31, characterized in that the composition further comprises an agent that induces modifications in a microbial phenotype.

33. COMPOSITION, according to any of the preceding claims, characterized by at least one bacterial additive comprising B. amyloliquefaciens, B. subtilis, B. pumilus and B. licheniformis in a ratio of 1:1:1:

1.

34. COMPOSITION, according to any one of claims 1 to 32, characterized by at least one bacterial additive Petition 870250089817, dated 02 / 10 / 2025, page 127 / 130 6 / 6 comprising B. amyloliquefaciens, B. pumilus and B. licheniformis in a 1:1:1 ratio.

35. COMPOSITION, characterized by comprising: a) a liquid component comprising the composition as defined in any of the preceding claims; and b) a carrier component comprising a mineral fertilizer or seed.

36. METHOD OF USING THE COMPOSITION, as defined in any of the preceding claims, characterized in that the method comprises a specialized use.

37. METHOD, according to claim 36, characterized by its specialized use comprising greater efficiency in the use of nutrients and / or availability of nutrients for crops.

38. METHOD, according to claim 37, characterized in that the nutrient comprises nitrogen, phosphorus, potassium, calcium, magnesium, sulfur, non-essential nutrients and / or micronutrients.

39. METHOD, according to claim 38, characterized in that the non-essential nutrients comprise titanium, selenium and / or silicon. Petition 870250089817, dated 02 / 10 / 2025, pp. 128 / 130

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