Fertilizer comprising elemental sulfur and sulfur-oxidizing microorganisms

Abstract

A fertilizer composition comprising a plurality of one or more live sulfur oxidation microorganisms, elemental sulfur, and a solid fertilizer comprising nitrogen, phosphorous, and / or potassium and optionally talc and methods for producing and using the same.

Description

DESCRIPTIONFERTILIZER COMPRISING ELEMENTAL SULFUR AND SULFUR-OXIDIZING MICROORGANISMSCROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to and the benefit of European Patent Application No. 24175244.3, filed May 10, 2024, the contents of which is incorporated into the present application by reference in its entirety.BACKGROUNDA. Field of the invention

[0002] The invention generally concerns fertilizer compositions containing elemental sulfur and sulfur-oxidizing microorganisms and methods of making the same. In particular embodiments, the fertilizer composition comprises a solid fertilizer comprising nitrogen, phosphorous, and / or potassium, elemental sulfur, and a plurality of live sulfur-oxidizing microorganisms.B. Description of related art

[0003] Soil nutrients, such as nitrogen, phosphorus, potassium, and sulfur, as well as trace elements such as iron, zinc, copper, and magnesium, are useful for achieving thriving agriculture and the growth of plants. Upon repeated planting cycles, the quantity of these nutrients in the soil may be depleted, resulting in reduced plant growth and decreased production. To counter this effect, fertilizers have been developed to help replace the depleted vital nutrients. Single-nutrient fertilizers and multi-nutrient fertilizers, such as fertilizer blends, have been developed to meet the varied needs of crop production worldwide.

[0004] Sulfur is classified as a secondary nutrient required for plant growth and development. However, recent studies suggest that sulfur is more essential than other secondary nutrients and it should be considered a primary nutrient. Plants typically cannot utilize elemental sulfur, but elemental sulfur can be converted to a more plant usable form, ionized sulfate (SCU)2-, by microorganism known as sulfur-oxidizing microorganisms. Sulfates can also be supplied in readymade sulfate products. Sulfur-oxidizing microorganisms are not active in a considerable amount of cultivated lands. For these lands, applying elemental sulfur is mostly useless and not recommended. Sulfate products can instead be used to providefor the sulfur needs of plants in these lands. However, sulfate products are expensive compared to elemental sulfur products.

[0005] The amount and variety of microorganisms, such as sulfur-oxidizing bacteria, can vary from place to place and can vary over time. Therefore, similar to the replenishment of nutrients in soil using fertilizers, soil may also need to be replenished with beneficial microorganisms, such as sulfur-oxidizing bacteria.

[0006] Currently, most supplemental microorganisms are delivered to the soil (i) directly (e.g., as a solid and / or liquid form application), (ii) in solution with irrigation water, (iii) coated on seeds, or (iv) coated on fertilizers. These delivery methods are limited in scope and the delivery methods and suitable carriers for the microorganisms limit the type of microorganisms that can be feasibly utilized commercially. For example, high temperatures associated with fertilizer creation can render most microorganisms inactive and / or inefficient and highly acidic or basic carriers, such as many fertilizers, can be harmful to microorganisms by contact. There exists a need to be able to widen the scope of microorganisms used and / or widen the scope of microorganism delivery methods.SUMMARY

[0007] A discovery has been made that provides a solution to at least some of the problems discussed above. In particular, disclosed herein are microbe-enhanced fertilizers comprising: nitrogen, phosphorous, and / or potassium; elemental sulfur; and a plurality of one or more live sulfur-oxidizing microorganisms. The sulfur-oxidizing microorganism may be in some instances, bacteria of a Starkeya genus and / or a Thibacillus genus. Non-limiting examples include the use of Starkeya novella as a sulfur-oxidizing microorganism. In some instances, the microbe-enhanced fertilizer also contains talc. Further, the microbe-enhanced fertilizer composition may enable the delivery of stable microorganisms along with each granule of fertilizer. The fertilizer granule can be delivered where and when the microorganism is needed to convert elemental sulfur into a plant-consumable sulfate form as a quick release fertilizer and / or over time as the fertilizer degrades as a slow release fertilizer. Additionally, the microbe- enhanced fertilizer composition can have an increased microorganism and / or fertilizer shelflife as compared to other microorganism containing fertilizers.

[0008] In some aspects, the fertilizer composition comprises or consists of a plurality of one or more live bacteria of a Starkeya genus and / or a Thibacillus genus, talc, elemental sulfur, and a solid fertilizer comprising nitrogen, phosphorous, and / or potassium.

[0009] In some aspects, the fertilizer composition comprises microorganisms. In some instances, the microorganisms are a purple sulfur bacteria or green sulfur bacteria. In some instances, the microorganisms are sulfur-oxidizing microorganisms. In some instances, the microorganisms comprise one or more live bacteria of a Starkeya genus and / or a Thibacillus genus. In some instances, the live bacteria comprises Starkeya novella. In some aspects, the live microorganism comprises or further comprises Acidithiobacillus ferroxidans. Cytobacillus firmus, Psedomonas stutzeri, and / or Enterobacter luduigii. In some aspects, the live microorganism oxidizes the elemental sulfur into a plant-consumable sulfate (SO4)2’ form as a sulfur source.

[0010] In certain aspects, the fertilizer composition comprises elemental sulfur. In some aspects, the elemental sulfur comprises micronized sulfur.

[0011] In some aspects, at least a portion of the microorganisms, the talc, and / or the elemental sulfur are comprised in a coating on the solid fertilizer. In some aspects, any one of a solid fertilizer, talc, and / or elemental sulfur may be at least partially surrounded by the microorganisms. In some aspects, the microorganisms are at least partially surrounded with the talc and / or the elemental sulfur. In some aspects, at least a portion of the solid fertilizer is at least partially coated by the microorganisms, the elemental sulfur, and / or the talc. In some aspects, at least a portion of the microorganisms are in contact with the solid fertilizer, elemental sulfur, and / or the talc. In some instances, the solid fertilizer, the talc, and / or the microorganisms are coated on the elemental sulfur. In some instances, the solid fertilizer, the elemental sulfur, and / or the microorganisms are coated on the talc. In some instances, the coating comprises the solid fertilizer, the elemental sulfur, the talc, and / or the microorganisms in separate layers in the coating, such as two or three layers. In some instances, the coating contains the solid fertilizer, the elemental sulfur, the talc, and / or the microorganisms in a single layer, such as a homogenous layer.

[0012] In some aspects, the fertilizer composition further comprises an adhesive agent, such as a water-soluble adhesive agent. In some aspects, the water-soluble adhesive agent comprises glycerol, carboxy methyl cellulose (CMC), polyvinyl pyrrolidone (PVP), gum arabic, guar gum, monosaccharides of CMC, monosaccharides of arabic gum, monosaccharides of guar gum, disaccharides of CMC, disaccharides of arabic gum, disaccharides of guar gum, or combinations thereof. In some aspects, the composition comprises 0.5-2 wt.% of the adhesive agent. In some aspects, the composition further comprises secondary nutrients such as calcium and / or magnesium. In some aspects, the composition contains trace elements, such as boronand / or zinc. In some aspects, the composition further comprises a stabilizer, such as starch, agar, polyethylene glycol, polyvinyl alcohol, ethanol, or combinations thereof. In some aspects, the composition contains a carrier, such as biochar, carbon black, talc, and / or a polysaccharide. In some aspects the adhesive agent, the stabilizer, and / or the carrier protect the microorganisms from contacting the solid fertilizer and / or elemental sulfur. In some instances, the adhesive agent, carrier, and / or stabilizer are comprised in a coating on the fertilizer, a coating of the microorganisms, and / or a layer of a coating.

[0013] In certain aspects, the fertilizer composition contains at least any one of, at most any one of, equal to any one of, or between any two of 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 wt.% of the solid fertilizer comprising nitrogen, phosphorous, and / or potassium based on the total weight of the fertilizer composition. In some aspects, the fertilizer composition contains at least any one of, at most any one of, equal to any one of, or between any two of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 wt. % of talc based on the total weight of the fertilizer composition. In some aspects, the fertilizer composition contains at least any one of, at most any one of, equal to any one of, or between any two of 1, 2, 3, 4, or 5 wt. % of elemental sulfur based on the total weight of the fertilizer composition. In some aspects, the fertilizer composition contains at least any one of, at most any one of, equal to any one of, or between any two of 0.01, 0.03, 0.05, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95 or 1 wt. % of a plurality of microorganisms based on the total weight of the fertilizer composition.

[0014] In some aspects, the fertilizer is homogenous. In some aspects, the fertilizer composition is a solid fertilizer. In some aspects, the fertilizer composition is a slurry. In some aspects, the fertilizer is a liquid. In some aspects, the fertilizer composition is capable of being, or is, blended with other fertilizers. In some aspects, the fertilizer composition is capable of being used in fertigation. In some aspects, the fertilizer composition is capable of being used in a foliar spray.

[0015] Certain aspects are directed to a method of making the fertilizer composition, including any fertilizer described herein. The method can include coating an outer surface of a core comprising the solid fertilizer, the elemental sulfur, and / or the talc with the solid fertilizer, the elemental sulfur, the talc, and / or the microorganism. In some instances, the coating is formed by contacting the outer surface of the core with a solution comprising the solid fertilizer, the elemental sulfur, the talc, and / or the microorganisms. In some instances, the coating is formed by contacting the outer surface of the core with an adhesive agent, such as a water-soluble adhesive agent, before contact with the coating. In some instances, the method comprises combining the microorganisms and the talc before contacting the microorganism and the talc with the fertilizer and / or the elemental sulfur. In some instances, the method comprises coating the solid fertilizer, the elemental sulfur, the talc, and / or the microorganisms in separate layers in the coating, such as two or three layers. In some instances, the method comprises coating the solid fertilizer, the elemental sulfur, the talc, and / or the microorganisms in a single layer, such as a homogenous layer. In some instances, the method comprises combining and mixing the solid fertilizer with the elemental sulfur, the talc, and / or the microorganism to form a homogenous composition.

[0016] In some embodiments, methods of fertilizing are described. A method can include applying a fertilizer composition, and / or blended or compounded fertilizer composition described herein to a portion of a soil, a crop, water, or any combination of the soil, the water, and the crop. In some embodiments, the soil is at least partially or fully submerged under water (e.g., rice paddy crops) and the granules sink in the water to contact the soil.

[0017] Certain embodiments of the present invention are characterized through the following aspects.

[0018] Aspect 1 concerns a fertilizer composition comprising: a plurality of one or more live bacteria of a Starkeya genus and / or a Thibacillus genus; talc; elemental sulfur; and a solid fertilizer comprising nitrogen, phosphorous, and / or potassium.

[0019] Aspect 2 concerns the fertilizer composition of aspect 1, wherein the bacteria comprises Starkeya novella.

[0020] Aspect 3 concerns the fertilizer composition of any one of aspects 1 to 2, wherein the elemental sulfur comprises micronized sulfur.

[0021] Aspect 4 concerns the fertilizer composition of any one of aspects 1 to 3, wherein the bacteria further comprises Acidithiobacillus ferroxidans, Cytobacillus firmus, Psedomonas stutzeri, and / or Enterobacter luduigii.

[0022] Aspect 5 concerns the fertilizer composition of any one of aspects 1 to 4, wherein at least a portion of the bacteria, the talc, and / or the elemental sulfur are comprised in a coating on the solid fertilizer.

[0023] Aspect 6 concerns the fertilizer composition of any one of aspects 1 to 5, wherein the composition further comprises a water-soluble adhesive agent.

[0024] Aspect 7 concerns the fertilizer composition of aspect 6, wherein the water-soluble adhesive agent comprises glycerol, carboxy methyl cellulose (CMC), polyvinyl pyrrolidone (PVP), gum arabic, guar gum, monosaccharides of CMC, monosaccharides of arabic gum, monosaccharides of guar gum, disaccharides of CMC, disaccharides of arabic gum, disaccharides of guar gum, or combinations thereof.

[0025] Aspect 8 concerns the fertilizer composition of aspect 7, wherein the composition comprises 0.5-2 wt.% of the water-soluble adhesive agent.

[0026] Aspect 9 concerns the fertilizer composition of any one of aspects 1 to 8, wherein the composition further comprises: secondary nutrients comprising calcium and / or magnesium; and / or trace elements comprising boron and / or zinc.

[0027] Aspect 10 concerns the fertilizer composition of any one of aspects 1 to 9, wherein the composition further comprises: a stabilizer comprises starch, agar, polyethylene glycol, polyvinyl alcohol, ethanol, or combinations thereof; and / or a carrier comprising biochar and / or a polysaccharide.

[0028] Aspect 11 concerns the fertilizer composition of any one of aspects 1 to 10, wherein the composition comprises 90 to 98 wt.% of the solid fertilizer, 1 to 8 wt.% of the talc, 1 to 3 wt.% of the elemental sulfur, and / or 0.05 to 0.5 wt. % of the plurality of bacteria based on the total weight of the fertilizer composition.

[0029] Aspect 12 concerns the fertilizer composition of any one of aspects 1 to 11, wherein at least a portion of the bacteria are at least partially surrounded by the elemental sulfur and / or the talc.

[0030] Aspect 13 concerns the fertilizer composition of any one of aspects 1 to 12, wherein at least a portion of the solid fertilizer is at least partially coated by the bacteria, the elemental sulfur, and / or the talc.

[0031] Aspect 14 concerns the fertilizer composition of any one of aspects 1 to 12, wherein the fertilizer is homogenous.

[0032] Aspect 15 concerns a method of making the fertilizer composition of any one of aspects 1 to 13, the method comprising coating an outer surface of a core comprising the solid fertilizer with the elemental sulfur, the talc, and / or the bacteria.

[0033] Aspect 16 concerns the method of aspect 15, wherein the coating is formed by contacting the outer surface of the core with a solution comprising the elemental sulfur, the talc, and / or the bacteria.

[0034] Aspect 17 concerns the method of any one of aspects 15 to 16, wherein the coating is formed by contacting the outer surface of the core with a water-soluble adhesive agent prior to contact with the elemental sulfur, the talc, and / or the bacteria.

[0035] Aspect 18 concerns the method of any one of aspects 15 to 17, further comprising combining the bacteria and the talc before contacting the bacteria and the talc with the fertilizer.

[0036] Aspect 19 concerns a method of making the fertilizer composition of aspect 14, the method comprising combining and mixing the solid fertilizer with the elemental sulfur, the talc, and / or the bacteria to form a homogenous composition.

[0037] Aspect 20 concerns a method of fertilizing, the method comprising applying the fertilizer composition of any one of the aspects 1 to 14 to a soil, a crop, water, or any combination thereof.

[0038] The following includes definitions of various terms and phrases used throughout this specification.

[0039] The term “fertilizer” is defined as a material applied to soils or to plant tissues to supply one or more plant nutrients essential or beneficial to the growth of plants and / or stimulants or enhancers to increase or enhance plant growth. Non-limiting examples of fertilizers include materials having one or more of urea, ammonium nitrate, calcium ammonium nitrate, urea calcium sulfate adduct, one or more superphosphates, binary NP fertilizers, binary NK fertilizers, binary PK fertilizers, NPK fertilizers, molybdenum, zinc, copper, boron, cobalt, and / or iron. In some embodiments, fertilizers include agents thatenhance plant growth and / or enhance the ability for a plant to receive the benefit of a fertilizer, such as, but not limited to bio stimulants, urease inhibitors, and nitrification inhibitors.

[0040] The term “microbe” or “microorganism” can include bacteria, fungi, protists, and / or archaea.

[0041] The term “micronutrient” is defined as a chemical element or substance used in trace amounts for the normal growth and development of a plant. Non-limiting examples of micronutrients include B, Cu, Fe, Mn, Mo, Zn, Se, and Si or compounds thereof.

[0042] The term “secondary nutrient” is defined as a chemical element or substance used in moderate amounts for plant growth and are less likely to limit crop growth in comparison to N, P, and K. Non-limiting examples of secondary nutrients include Ca, Mg, and S.

[0043] The term “organic agent” is defined as a substance that is produced by or part of an organism. Non-limiting examples of organic agents suitable for fertilizer include neem oil, seaweed extract, bio- stimulants, char, bio waste, ashes from incineration of animal waste or animal tissues, and diatomaceous earth.

[0044] The term “granule” can include a solid material. A granule can have a variety of different shapes, non-limiting examples of which include a spherical, a puck, an oval, a rod, an oblong, or a random shape.

[0045] The terms “about” or “approximately” are defined as being close to as understood by one of ordinary skill in the art. In one non-limiting embodiment, the terms are defined to be within 10%, preferably within 5%, more preferably within 1%, and most preferably within 0.5%.

[0046] The terms “wt.%,” “vol.%,” or “mol.%” refers to a weight percentage of a component, a volume percentage of a component, or molar percentage of a component, respectively, based on the total weight, the total volume of material, or total moles, that includes the component. In a non-limiting example, 10 grams of component in 100 grams of the material is 10 wt.% of component.

[0047] The term “substantially” and its variations are defined to include ranges within 10%, within 5%, within 1%, or within 0.5%.

[0048] The terms “inhibiting” or “reducing” or “preventing” or “avoiding” or any variation of these terms, when used in the claims and / or the specification, includes any measurable decrease or complete inhibition to achieve a desired result.

[0049] The term “effective,” as that term is used in the specification and / or claims, means adequate to accomplish a desired, expected, or intended result.

[0050] The use of the words “a” or “an” when used in conjunction with any of the terms “comprising,” “including,” “containing,” or “having” in the claims, or the specification, may mean “one,” but it is also consistent with the meaning of “one or more,” “at least one,” and “one or more than one.”

[0051] The words “comprising” (and any form of comprising, such as “comprise” and “comprises”), “having” (and any form of having, such as “have” and “has”), “including” (and any form of including, such as “includes” and “include”), or “containing” (and any form of containing, such as “contains” and “contain”) are inclusive or open-ended and do not exclude additional, unrecited elements or method steps.

[0052] The microbe-enhanced fertilizer composition and methods of producing the microbe-enhanced fertilizer composition of the present invention can “comprise,” “consist essentially of,” or “consist of’ particular ingredients, components, compositions, steps, etc. disclosed throughout the specification. With respect to the transitional phase “consisting essentially of,” in one non-limiting aspect, a basic characteristic of the fertilizer of the present invention is the presence of a microorganism in or on a granulated fertilizer.BRIEF DESCRIPTION OF THE DRAWINGS

[0053] Advantages of the present invention may become apparent to those skilled in the art with the benefit of the following detailed description and upon reference to the accompanying drawings.

[0054] FIG. 1 A and FIG. IB depicts an exemplary homogenous microbe-enhanced fertilizer composition (A) or a coated microbe-enhanced fertilizer composition (B).

[0055] FIG. 2 depicts an exemplary microbe-enhanced fertilizer production method.

[0056] FIG. 3A and FIG. 3B depicts an exemplary homogenous microbe-enhanced liquid inoculant composition left or a urea granule directly coated with the microbe-enhanced liquid inoculant (right) at the time of coating (A) or after 5 days of storage (B).

[0057] While the invention is susceptible to various modifications and alternative forms, specific embodiments thereof are shown by way of example in the drawings. The drawings may not be to scale.DETAILED DESCRIPTION

[0058] Disclosed herein, among other things, is a microbe-enhanced fertilizer and a method of producing a microbe-enhanced fertilizer. The microbe-enhanced fertilizer composition enables the delivery of microorganisms along with each granule of fertilizer, where and / or where the microorganism is needed to transform the elemental sulfur into a plant-consumable sulfate form and / or over time as the fertilizer degrades. Additionally, the microbe-enhanced fertilizer composition can have an increased microorganism and / or fertilizer shelf-life.

[0059] The methods and / or compositions of the current disclosure provide an economically efficient means to produce and / or utilize a stable and high-quality microbe-enhanced fertilizer. These and other non-limiting aspects of the present invention are discussed in further detail in the following sections.A. Fertilizer Compositions

[0060] In certain aspects, the fertilizer composition contains at least any one of, at most any one of, equal to any one of, or between any two of 80 wt. %, 81 wt. %, 82 wt. %, 83 wt. %, 84 wt. %, 85 wt. %, 86 wt. %, 87 wt. %, 88 wt. %, 89 wt. %, 90 wt. %, 91 wt. %, 92 wt. %, 93 wt. %, 94 wt. %, 95 wt. %, 96 wt. %, 97 wt. %, 98 wt. %, or 99 wt.% of the solid fertilizer comprising nitrogen, phosphorous, and / or potassium, at least any one of, at most any one of, equal to any one of, or between any two of 1 wt. %, 2 wt. %, 3 wt. %, 4 wt. %, or 5 wt. % of the elemental sulfur, and / or at least any one of, at most any one of, equal to any one of, or between any two of 0.01, 0.03, 0.05, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95 or 1 wt. % of the plurality of microorganism based on the total weight of the fertilizer composition based on the total weight of the fertilizer composition. In some aspects, the fertilizer composition contains at least any one of, at most any one of, equal to any one of, or between any two of 1 wt. %, 2 wt. %, 3 wt. %, 4 wt. %, 5 wt. %, 6 wt. %, 7 wt. %, 8 wt. %, 9 wt. %, or 10 wt. % of the talc.

[0061] In some aspects, the fertilizer composition comprises or consists of a plurality of one or more live bacteria of a Starkeya genus and / or a Thibacillus genus, elemental sulfur, and a solid fertilizer comprising nitrogen, phosphorous, and / or potassium, and optionally talc. In some aspects, the solid fertilizer comprising urea, di-ammonium phosphate (DAP), mono ammonium phosphate (MAP), or any other fertilizer comprising nitrogen, phosphorous, and / or potassium.

[0062] In some aspects, the fertilizer composition comprises microorganisms. In some aspects, the microorganisms comprise a plurality of one or more live microorganism that is capable of oxidizing the elemental sulfur into a plant-consumable sulfate (SO4)2’ form as a sulfur source. In some aspects, the elemental sulfur comprises micronized sulfur.

[0063] In some aspects, at least a portion of the microorganism, the talc, and / or the elemental sulfur are comprised in a coating on the solid fertilizer. In some aspects, at least a portion of the microorganism, the elemental sulfur, and / or the solid fertilizer are comprised in a coating on the talc. In some aspects, at least a portion of the microorganism, the talc, and / or the solid fertilizer are comprised in a coating on the elemental sulfur. In some aspects, any one of a solid fertilizer, talc, and / or elemental sulfur may be at least partially surrounded by the microorganism. In some aspects, the microorganism is at least partially surrounded with the talc, and / or the elemental sulfur. In some aspects, at least a portion of the solid fertilizer is at least partially coated by the microorganism, the elemental sulfur, and / or the talc. In some aspects, at least a portion of the microorganism are at least partially surrounded by the elemental sulfur and / or the talc.

[0064] In some aspects, the fertilizer composition further comprises an adhesive agent, such as a water-soluble adhesive agent. In some aspects, the water-soluble adhesive agent comprises glycerol, carboxy methyl cellulose (CMC), polyvinyl pyrrolidone (PVP), gum arabic, guar gum, monosaccharides of CMC, monosaccharides of arabic gum, monosaccharides of guar gum, disaccharides of CMC, disaccharides of arabic gum, disaccharides of guar gum, or combinations thereof. In some aspects, the composition comprises 0.5-2 wt.% of the adhesive agent. In some aspects, the composition further comprises secondary nutrients such as calcium and / or magnesium. In some aspects, the composition contains trace elements, such as boron and / or zinc. In some aspects, the composition further comprises a stabilizer, such as starch, agar, polyethylene glycol, polyvinyl alcohol, ethanol, or combinations thereof. In some aspects, the composition contains a carrier, such as biochar, carbon black, talc, and / or a polysaccharide. In some aspects the adhesive agent, the stabilizer, and / or the carrier protect the microorganisms from contacting the solid fertilizer and / or elemental sulfur. In some instances, the adhesive agent, carrier, and / or stabilizer are comprised in a coating on the fertilizer, a coating of the microorganisms, and / or a layer of a coating.

[0065] In some aspects, the fertilizer composition may contain 4 to 46 wt. % of nitrogen or at least any one of, at most any one of, equal to any one of, or between any two of 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33,34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, or 46 wt. % of nitrogen. In some aspects, the fertilizer composition may contain 5 to 35 wt. % of phosphorous or at least any one of, at most any one of, equal to any one of, or between any two of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 wt. % of phosphorous. In some aspects, the fertilizer composition may contain 5 to 35 wt. % of potassium or at least any one of, at most any one of, equal to any one of, or between any two of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 wt. % of potassium. In some aspects, the fertilizer composition may contain 0.5 to 10 wt. % or at least any one of, at most any one of, equal to any one of, or between any two of 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 wt.% of elemental sulfur. In some aspects, elemental sulfur includes micronized sulfur.

[0066] In some aspects, the composition does not contain or is substantially free of a compost, humic acid, Mg, Br, Fe, urea, ammonium nitrate, ammonium phosphate, diammonium phosphate, sulfate of ammonia, muriate of potash, potassium sulfate, magnesium carbonate, silicon, potassium nitrate, phosphate rock, lime, zeolite, hydrochloric acid, leopardite, torrefied organic waste comprising labile carbon, charcoal, urea, and / or calcium cyanamide.

[0067] The fertilizer can be of any suitable shape. Non-limiting shapes include spherical, cuboidal, cylindrical, puck shape, oval, and oblong shapes. In some aspects, the core can be of cylindrical shape with a circular, elliptical, ovular, triangular, square, rectangular, pentagonal, or hexagonal cross section, although cylindrical shaped core having a cross-section of other shapes can also be made. In some aspects, the core can have a dimension such as length, width, height and / or cross-sectional diameter between 0.5 mm to 5 mm or at least any one of, equal to any one of, or between any two of 0.5 mm, 1 mm, 1.5 mm, 2 mm, 2.5 mm, 3 mm, 3.5 mm, 4 mm, 4.5 mm, and 5 mm. In some particular aspects, the core can have a substantially spherical shape with an average diameter 1 mm to 5 mm or at least any one of, equal to any one of, or between any two of 0.5 mm, 1 mm, 1.5 mm, 2 mm, 2.5 mm, 3 mm, 3.5 mm, 4 mm, 4.5 mm, and 5 mm.

[0068] Referring to FIG. 1 a fertilizer granule 100 according to an example of the present invention is shown. The fertilizer granule (100) can contain a major portion of the solid fertilizer comprising nitrogen, phosphorous, and / or potassium (101) in the core, and a minor portion of the talc (102), elemental sulfur (103), and a plurality of live microorganisms such as one or more live bacteria of a Starkeya genus and / or a Thibacillus genus (104). In Fig. 1A, thesolid fertilizer, talc, elemental sulfur, and microorganisms (101, 102, 103, and 104) are uniformly distributed throughout the fertilizer granules. In some instances an adhesive agent, carrier, and / or stabilizer is also uniformly distributed throughout the fertilizer granules. In some instances, the fertilizer granule is homogenous. In some instances, the fertilizer granule is not homogenous. In some instances, the fertilizer granule is coated with a coating. In some instances, the core is a solid fertilizer (101) and is coated with a coating containing any one or a combination of talc, elemental sulfur, and / or microorganisms (102, 103, and 104). In some instances, the outer surface of the core is coated with an adhesive agent, carrier, and / or stabilizer (105) before contact with the elemental sulfur, the talc, and / or the microorganism (102, 103, and 104) as in Fig. IB. In some instances, the fertilizer granules or the fertilizer compositions further comprise other fertilizers, micronutrients, primary nutrients, additional urea, additional nitrogen nutrients, insecticides, herbicides, or fungicides, or combinations thereof.

[0069] The fertilizer granules can have desirable physical properties such as desired levels of abrasion resistance, granule strength, pelletizability, hygroscopicity, granule shape, and size distribution, which are important properties for the fertilizer core.

[0070] The fertilizer granules described herein can be comprised in a composition useful for application to soil, water, and / or a crop. In addition to the fertilizer granules, the composition may include other fertilizer compounds, micronutrients, primary nutrients, urea, nitrogen nutrients, insecticides, herbicides, or fungicides, or combinations thereof.

[0071] The fertilizer granules described herein can also be included in a blended composition comprising other fertilizer granules. The other fertilizer granules can be granules of urea, monoammonium phosphate (MAP), diammonium phosphate (DAP), muriate of potash (MOP), monopotassium phosphate (MKP), triple super phosphate (TSP), rock phosphate, single super phosphate (SSP), ammonium sulfate, and the like.B. Microorganisms

[0072] In some embodiments, a microorganism that can be included in a microbe-enhanced fertilizer can be a bacteria, archaea, fungi, or a protist. In some instances, the microorganism can include or can further include an endophyte, rhizosphere microbe, and / or phyllosphere microbe. In some embodiments, the microorganism can be included more than one species of microorganism. In some embodiments, more than one species, genus, phylum, class, and / orkingdom can be represented in a group of microorganisms protected and / or included in a microbe-enhanced fertilizer.

[0073] In some embodiments, a microorganism can be cultured and / or grown in a laboratory. In some embodiments, a microorganism can be obtained from a natural source. In some embodiments, a microorganism can be a spore / cyst-forming microorganism, such as a spore / cyst-forming bacteria. In some embodiments, a microorganism can be concentrated before addition to the fertilizer.

[0074] In some embodiments, a microorganism can be but is not limited to, one or more live bacteria of a Starkeya genus and / or a Thibacillus genus, sulfur-oxidizing bacterium, (e.g., an organism that can convert elemental sulfur into sulfate through the process of oxidation), phosphorus solubilizing, nitrite oxidizing, Nitrospira species, Methylobacterium species, and / or pink pigmented facultative methylotrophs (PPFM-trophs). In some embodiments, bacteria further comprises Acidithiobacillus ferroxidans, Cytobacillus firmus, Psedomonas stutz.eri. and / or Enterobacter luduigii.

[0075] In some embodiments, a microorganism can be induced to form spores / cysts prior to addition to the fertilizer. In some embodiments, a microorganism is not induced to form spores / cysts prior to and / or as part of addition to the fertilizer. In some embodiments, a microorganism is not chemically induced to form spores / cysts prior to and / or as part of addition to the fertilizer. In some embodiments, a microorganism has been selected for heat tolerance. In some embodiments, a microorganism has not been selected for heat tolerance.

[0076] In some embodiments, microorganisms can be cultured and / or concentrated to greater than, or equal to, approximately 104-1013cells per mL prior to addition to the fertilizer. In some embodiments, microorganisms can be cultured and / or concentrated to greater than, or equal to, approximately 108-109cells per mL prior to addition to the fertilizer.

[0077] Prior to contacting with a fertilizer to obtain a microbe-enhanced fertilizer, microorganisms may be protected. In some embodiments, microorganism protection may comprise any one or more of encapsulation, physical protection, and / or engineering methods. In some embodiments, microorganism protection is by contact with a protectant. In some instances, microorganism protection is by encapsulation in a protectant. In some embodiments, a microorganism is protected by addition of one or more physical protectants, engineering methods, encapsulating agents, water-soluble additives, adhesives, carriers, stabilizer additives, and / or dispersants.

[0078] In some embodiments, microorganism protection can comprise encapsulation with a stabilizer protectant. In some embodiments, a stabilizer comprises one or more of clay, diatomaceous earth, starch, agar, alginate, chitosan, polyethylene glycol (PEG), polyvinyl alcohol (PVA), polyacrylic acid, ethanol, humic acid, humates, talc, clay, peat, lignite, vermiculite, perlite and / or chemically modified versions of the same.

[0079] In some embodiments, chemical modification of a stabilizer can comprise, but is not limited to, one or more of esterification, alkylation, acetylation, phosphorylation, hydrophobic modification, sulfation, sulfomethylation, methylation, amidation, amination, protonation, halogenation, nitration, copolymerization, and / or physical or covalent cross -linking.

[0080] In some embodiments, the protection of a microorganism can also comprise the addition of an adhesive agent, such as a water-soluble adhesive agent. In some embodiments, a water-soluble adhesive agent can be, but is not limited to, glycerol, carboxy methyl cellulose (CMC), polyvinyl pyrrolidone (PVP), gum Arabic, guar gum, and / or mono and / or disaccharide based CMC / Arabic gum / guar gum.

[0081] In some embodiments, microorganism protection can comprise improved stickiness, stabilization, and surfactant and dispersal abilities. In some embodiments, such characteristics can be provided by protectants / inducers and nutrients (e.g. alginates / glycerol / PVA, PEG / polyvinylpyrrolidone (PVP), clay / humate, mono and disaccharides, CMC (carboxymethyl cellulose) / arabic gum / guar gum).

[0082] In some embodiments, protection can comprise the inclusion of certain stabilizers and / or additives at set proportions, including but not limited to 1:0.05, 1:0.10, 1:0.15, 1:0.20, 1:0.25, 1:0.30, 1:0.35, 1:0.40, 1:0.45, 1:0.50, 1:0.55, 1:0.60, 1:0.65, 1:0.70, 1:0.75, 1:0.80, 1:0.85, 1:0.90, 1:0.95, 1: 1, 1: 1.05, 1: 1.10, 1: 1.15, 1: 1.20, 1: 1.25, 1: 1.30, 1: 1.35, 1: 1.40, 1: 1.45, 1: 1.5, 1: 1.55, 1: 1.60, 1: 1.65, 1: 1.70, 1: 1.75, 1: 1.80, 1: 1.85, 1: 1.90, 1: 1.95, 1:2, 1:3, 1:4, 1:5, 1: 10, 1:20, 1:30, 1:40, 1:50, 1:60, 1:70, 1:80, 1:90, 1: 100, 1: 1,000, 1:10,000, 1: 100,000, 1: 1,000,000, or any range derivable therein.

[0083] In some embodiments, microorganisms can be cultured and / or concentrated to greater than, or equal to, approximately 104-1013cells per mL prior to addition to the fertilizer. In some embodiments, microorganisms can be cultured and / or concentrated to greater than, or equal to, approximately 108-109cells per mL prior to addition to the fertilizer.

[0084] In some embodiments, physical protection and / or engineering methods facilitate pelleting and / or layering of microorganisms as a liquid solution at the core or around a core ofa fertilizer granule. In some embodiments, a bolus of concentrated microbes in a liquid carrier are protected with a soluble additive, such as in a slurry.

[0085] In some embodiments, physical protection of microorganisms may comprise addition of protectants that are molecules and / or enzymes derived from thermophiles (e.g., heat tolerant microorganism), these molecules and / or enzymes may contribute to the thermoprotection phenotypes observed in thermophiles (see e.g., Pedro Lamosa et al., Thermo stabilization of Proteins by Diglycerol Phosphate, a New Compatible Solute from Hyperthermophile Archaeoglobus fulgidus. Applied and Environmental Microbiology, Vol. 66, No. 5, 01 May 2000). Molecules and / or enzymes derived from thermophiles include proteins, lipids, saccharides, nucleic acids, small molecules, and / or inorganic compounds. Thermophiles may include bacteria, archaea, protists, and / or fungi. Thermophile microorganisms may include microorganisms that can thrive, divide, and / or survive at temperatures of 50 °C or greater. A non-limiting example of a thermophile is Archaeoglobus fulgidus.

[0086] In some embodiments, engineering methods for the protection of microorganisms may comprise spray drying and / or freeze-drying of the microorganisms. Freeze-drying can be performed by freezing the microorganisms or a composition containing the microorganisms, exposing the frozen microorganism or composition containing the microorganism to pressures below atmospheric pressures, and removing ice from or surrounding the frozen microorganism or composition. The composition containing the microorganism can contain, in some instances, a cryoprotectant, encapsulating agent, water-soluble additive, stabilizer additive, and / or a dispersant.

[0087] In some embodiments, a microorganism is contacted with a protectant using a spray, liquid stream, semi-solid, or solid (such as a powder) comprising said protectant. In some embodiments, a protectant is contacted with a microorganism using a spray, liquid stream, semi-solid, or solid (such as a powder) comprising said microorganism.

[0088] In some embodiments, a protected microorganism is concentrated (e.g., settlement, centrifugation, affinity capture, selective growth media, etc.,) before contact with a protectant and / or prior to protecting. In some embodiments, a protected microorganism is contacted with the protectant or with the fertilizer at a concentration of higher than 1012cells per gram of the protectant. In some embodiments, a protectant is comprised in a liquid, suspension, and / or dried powder.

[0089] In some embodiments, a protected microorganism can contain low amounts of moisture. In some embodiments, the free-moisture content of a protected microorganism can be less than 0.6 wt.%, less than 0.5 wt.% water or 0.25 wt.% to less than 0.6 wt.% water. In some instances, the free moisture content is 0.5, 0.4, 0.3, 0.2, 0.1, or 0 wt.%.C. Methods of Making Fertilizer Compositions

[0090] A method of producing the fertilizer composition comprising a plurality of one or more live microorganism, elemental sulfur, and a solid fertilizer comprising nitrogen, phosphorous, and / or potassium and optionally talc is disclosed. In some aspects, the method can include coating an outer surface of a core comprising the solid fertilizer, the elemental sulfur, and / or the talc with the solid fertilizer, the elemental sulfur, the talc, and / or the microorganism. In some instances, the coating is formed by contacting the outer surface of the core with a solution comprising the solid fertilizer, the elemental sulfur, the talc, and / or the microorganisms. In some instances, the coating is formed by contacting the outer surface of the core with an adhesive agent before contact with the coating. In some instances, the method comprises combining the microorganisms and the talc before contacting the microorganism and the talc with the fertilizer and / or the elemental sulfur. In some instances, the method comprises coating the solid fertilizer, the elemental sulfur, the talc, and / or the microorganisms in separate layers in the coating, such as two or three layers. In some instances, the method comprises coating the solid fertilizer, the elemental sulfur, the talc, and / or the microorganisms in a single layer, such as a homogenous layer. In some instances, the method comprises combining and mixing the solid fertilizer with the elemental sulfur, the talc, and / or the microorganism to form a homogenous composition.

[0091] In some aspects, the fertilizer composition can contain 0 wt. % to 2 wt. % or at least any one of, equal to any one of, or between any two of 0 wt. %, 0.1 wt. %., 0.15 wt. %, 0.2 wt. %, 0.25 wt. %, 0.3 wt. %, 0.35 wt. %, 0.4 wt. %, 0.45 wt. %, 0.5 wt. %, 0.55 wt. %, 0.6 wt. %, 0.65 wt. %, 0.7 wt. %, 0.75 wt. %, 0.8 wt. %, 0.85 wt. %, 0.9 wt. %, 0.95 wt. %, 1 wt. %, 1.1 wt. %., 1.15 wt. %, 1.2 wt. %, 1.25 wt. %, 1.3 wt. %, 1.35 wt. %, 1.4 wt. %, 1.45 wt. %, 1.5 wt. %, 1.55 wt. %, 1.6 wt. %, 1.65 wt. %, 1.7 wt. %, 1.75 wt. %, 1.8 wt. %, 1.85 wt. %, 1.9 wt. %, 1.95 wt. %, and 2 wt. %, of water.

[0092] In some aspects, the sulfur-oxidizing microorganism, talc, solid fertilizer, and / or elemental sulfur can be coated at 40 °C to 90 °C, or at least any one of, equal to any one of, or between any two of 40 °C, 45 °C, 50 °C, 55 °C, 60 °C, 65 °C, 70 °C, 75 °C, 80 °C, 85 °C, and90 °C. In some aspects, combining the microorganism and the talc before coating the microorganism and the talc with the fertilizer. In certain aspects, the mixture can be dried at 35 °C, 40 °C, 45 °C, 50 °C, 55 °C, 60 °C, 65 °C, 70 °C, 75 °C, 80 °C, 85 °C, 90 °C, 95 °C, 100 °C or greater, or any temperature or range thereof or there between to form a dried mixture. The amount of water, e.g., moisture, in the dried mixture can be less than 0.6 wt.%, 0.5 wt.%, 0.4 wt.%, 0.3 wt.%, 0.2 wt.%, 0.1 wt.%, or less, or any amount or range thereof or there between.

[0093] In some instances, producing the fertilizer composition herein comprises contacting the solid fertilizer, elemental sulfur, and / or the talc mixture with the plurality of live sulfuroxidizing microorganisms and mixing to form a fertilizer paste. The fertilizer paste can then be pelletized, such as a pellet with a size of 2 x3 mm. The pellets may be dried at 65 °C to 95 °C, or at least any one of, equal to any one of, or between any two of 65, 70, 75, 80, 85, 90, and 95 °C for a period of time, such as 24 hours.

[0094] In some instances, coating comprises contacting the solid fertilizer with the elemental sulfur and / or the talc to form a mixture, then contacting the mixture with the plurality of live microorganisms such as sulfur-oxidizing bacteria and mixing. The pellets formed in this method may comprise at least a partial coating of live microorganisms such as sulfur-oxidizing bacteria on the solid fertilizer, elemental sulfur, and / or the talc mixture.

[0095] FIG. 2 shows a flow chart of a method 200 for making a fertilizer granule according to one example of the present invention. Referring to FIG. 2, a solid fertilizer comprising nitrogen, phosphorous, and / or potassium and its outer surface form a core 201 that is coated with a solution comprising the elemental sulfur 203, the talc 204, and / or the microorganism 205 to form a final fertilizer granule 206. In some instances, the coating is formed by contacting the outer surface of the core 201 and coating with a water-soluble adhesive agent 202 before contact with the elemental sulfur 203, the talc 204, and / or the microorganism 205. In some instances, the microorganism 205 and the talc 204 are combined before contacting the microorganism and the talc with the solid fertilizer 201.

[0096] In some embodiments, microbe-enhanced fertilizers of the present disclosure comprise any granulatable fertilizer (e.g., granular fertilizer). In some embodiments, a microbe- enhanced fertilizer comprises or excludes one or more granular fertilizers, preferably but not limited to, one or more of urea, single super phosphate (SSP), triple super phosphate (TSP), ammonium sulfate, monoammonium phosphate (MAP), diammonium phosphate (DAP),muriate of potash (MOP), sulfate of potash (SOP), potassium sulfate, binary NP fertilizers, binary NK fertilizers, binary PK fertilizers, and / or a nitrogen phosphorus potassium (NPK) mix.

[0097] In some embodiments, fertilizer granulation comprises chemically reacting reactants to form the fertilizer. In some embodiments, the fertilizer is formed from or is provided in a solution. In some embodiments, the fertilizer is formed or is provided in a fertilizer melt. The fertilizer melt can be formed, in some instances by evaporating a fertilizer solution. In some instances, the fertilizer is formed from or is provided in a solidified fertilizer. The solidified fertilizer can be formed, in some instances by cooling a fertilizer melt. In some instances, the fertilizer is formed from or is provided in a granulated fertilizer. The granulated fertilizer can be formed, in some instances by granulating the solidified fertilizer melt or a cooling fertilizer melt.

[0098] As described herein, in some embodiments, a microbe-enhanced fertilizer is produced when a microorganism is contacted with the fertilizer before or during granulation.

[0099] In some embodiments, a fertilizer dryer can be heated by steam, such as in a steam jacketed dryer. In some embodiments, a fertilizer dryer can be or can be part of a rotating dryer. In some embodiments, a granulator can be separate from a dryer. In some embodiments, a dryer and granulator can be the same vessel or part of a same vessel. In some embodiments, a granulator, may include a rotatable section, a rotatable internal container, and / or a section that vibrates. In some embodiments, the rotatable section and / or rotatable internal container may contain internal flights and / or be rotated to induce movement of a fertilizer composition in the granulator. In some embodiments, a granulator can be or can be part of a granulation drum, pugmill, pan granulator, etc.

[0100] In some embodiments, water or an aqueous solution, such as steam and / or a scrubber solution, can be combined with a fertilizer composition in a granulator to facilitate granulation of a fertilizer composition.

[0101] In some embodiments, a microorganism or a protected microorganism is contacted with a fertilizer by spraying onto a fertilizer particle and / or granule, by mixing into a fertilizer, by spraying a fertilizer onto the microorganism, by coating a fertilizer, by being coated by a fertilizer, by being encapsulated in a fertilizer matrix, by encapsulating a fertilizer to form a matrix of the microorganism, etc.

[0102] In some embodiments, a microorganism or a protected microorganism is contacted with a fertilizer using a spray, liquid stream, semi-solid, or solid (such as a powder) comprising said microorganism. In some embodiments, a microorganism is contacted with a fertilizer using a dosage pump or a spray head.

[0103] In some embodiments, a microorganism or a protected microorganism is concentrated (e.g., settlement, centrifugation, affinity capture, selective growth media, etc.,) prior to contact with a fertilizer to form a microbe-enhanced fertilizer. In some embodiments, the microorganism is contacted with the fertilizer at a concentration of 104-1012cells per gram of the fertilizer. In some preferred embodiments, a microorganism or a protected microorganism is contacted with the fertilizer at a concentration of 108-109cells per gram of the fertilizer. In some embodiments, a microorganism is comprised in a liquid, suspension, and / or dried powder.

[0104] In some embodiments, a microbe-enhanced fertilizer particle can have a crush strength of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 kgf / particle, or more, or any amount there between, preferably 2 kgf / particle to 5 kgf / particle.

[0105] In some embodiments, a microbe-enhanced fertilizer can contain a coating on the surface of one or more particles. In some instances, the coating can include nutrients for a plant, inhibitors of urea hydrolysis and / or nitrification, agents to slow or increase the rate of degradation of the granule and / or fertilizers, agents to repel moisture and / or provide a hydrophobic layer, agents that decrease or increase the reactivity of the granule and / or fertilizers, agents that provide additional benefits to plants, agents that increase the stability and / or crush strength of the granule and / or fertilizers, pH buffering agents, drying agents, etc. or any combination thereof. The coating can be a commercially available coating, an oil, a fertilizer, a micronutrient, talc, a seaweed and / or seaweed extract, a wax, etc. In some instances, the coating can contain surfactants. In some instances, the coating contains a wax, surfactants, and / or an amine-based compound.D. Methods of Using Microbe-enhanced Fertilizers

[0106] In some embodiments, microbe-enhanced fertilizer compositions of the present disclosure can be used in methods of increasing the amount of one or more nutrients and one or more microorganisms in soil, and of enhancing plant growth. In some embodiments, methods can include applying to the soil an effective amount of a composition microbe- enhanced fertilizers of the present disclosure. In some embodiments, methods may includeincreasing the growth and yield of crops, trees, ornamentals, etc. such as, for example, palm, coconut, rice, wheat, corn, barley, oats, and soybeans. In some embodiments, methods can include applying microbe-enhanced fertilizer of the present disclosure to at least one of a soil, an organism, a liquid carrier, a liquid solvent, etc. (e.g., a target substrate).

[0107] In some embodiments, a microbe-enhanced fertilizer can be stored. In some embodiments, the microbe-enhanced fertilizer can be stored for any amount of time, such as 1 minute, 10 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, 16 hours, 1 day, 2 days, 5 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 1 year, 2 years, 3 years, 4 years, 5 years or more, or any amount of time or range thereof or there between without 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43,44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68,69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93,94, 95, 96, 97, 98, 99, or 100 % of the microorganisms in the micro-enhanced fertilizer dying. In some embodiments, the microorganisms and / or fertilizer components of the microbe- enhanced fertilizer composition have an extended shelf life relative to microbe-enhanced fertilizers created through traditional methods.

[0108] In some embodiments, once a microbe-enhanced fertilizer is applied to a target substrate, microorganism protection materials degrade under field conditions and release the protected microorganisms to deliver their bio-effects.

[0109] Non-limiting examples of plants that can benefit from the microbe-enhanced fertilizer of the present invention include vines, trees, shrubs, stalked plants, fems, etc. The plants may include orchard crops, vines, ornamental plants, food crops, timber, and harvested plants. The plants may include Gymnosperms, Angiosperms, and / or Pteridophytes. The Gymnosperms may include plants from the Araucariaceae, Cupressaceae, Pinaceae, Podocarpaceae, Sciadopitaceae, Taxaceae, Cycadaceae, and Ginkgoaceae families. The Angiosperms may include plants from the Aceraceae, Agavaceae, Anacardiaceae, Annonaceae, Apocynaceae, Aquifoliaceae, Araliaceae, Arecaceae, Asphodelaceae, Asteraceae, Berberidaceae, Betulaceae, Bignoniaceae, Bombacaceae, Boraginaceae, Burseraceae, Buxaceae, Canellaceae, Cannabaceae, Capparidaceae, Caprifoliaceae, Caricaceae, Casuarinaceae, Celastraceae, Cercidiphyllaceae, Chrysobalanaceae, Clusiaceae, Combretaceae, Cornaceae, Cyrillaceae, Davidsoniaceae, Ebenaceae, Elaeagnaceae, Ericaceae, Euphorbiaceae, Fabaceae, Fagaceae, Grossulariaceae, Hamamelidaceae, Hippocastanaceae,Illiciaceae, Juglandaceae, Lauraceae, Lecythidaceae, Lythraceae, Magnoliaceae, Malpighiaceae, Malvaceae, Melastomataceae, Meliaceae, Moraceae, Moringaceae, Muntingiaceae, Myoporaceae, Myricaceae, Myrsinaceae, Myrtaceae, Nothofagaceae, Nyctaginaceae, Nyssaceae, Olacaceae, Oleaceae, Oxalidaceae, Pandanaceae, Papaveraceae, Phyllanthaceae, Pittosporaceae, Platanaceae, Poaceae, Polygonaceae, Proteaceae, Punicaceae, Rhamnaceae, Rhizophoraceae, Rosaceae, Rubiaceae, Rutaceae, Salicaceae, Sapindaceae, Sapotaceae, Simaroubaceae, Solanaceae, Staphyleaceae, Sterculiaceae, Strelitziaceae, Styracaceae, Surianaceae, Symplocaceae, Tamaricaceae, Theaceae, Theophrastaceae, Thymelaeaceae, Tiliaceae, Ulmaceae, Verbenaceae, and / or Vitaceae family.

[0110] In some embodiments, the effectiveness of compositions comprising microbe- enhanced fertilizers of the present invention can be ascertained by measuring the amount of particular nutrients in the soil at various times after applying the microbe-enhanced fertilizer composition to the soil. In some embodiments, the effectiveness of compositions comprising microbe-enhanced fertilizers of the present invention can be ascertained by measuring the amount of the microorganism in the soil at various times after applying the microbe-enhanced fertilizer composition to the soil. It is understood that different soils have different characteristics, which can affect the stability of nutrients and microorganisms in the soil. In some embodiments, the effectiveness of a microbe-enhanced fertilizer composition can be directly compared to other fertilizer compositions by doing a side-by-side comparison in the same soil under the same conditions.

[0111] In some embodiments, microbe-enhanced fertilizers according to the present disclosure can have a density that is greater than water. This can allow the granules and / or fertilizers to sink in water rather than float. This can be especially beneficial in instances where application is intended to a crop that is at least partially or fully submerged in water. A nonlimiting example of such a crop is rice, as the ground in a rice paddy is typically submerged in water. Thus, application of microbe-enhanced fertilizers to such crops can be performed such that the granules and / or fertilizer are homogenously distributed on the ground that is submerged under water.EXAMPLES

[0112] The present invention will be described in greater detail by way of specific examples. The following examples are offered for illustrative purposes only, and are not intended to limit the invention in any manner. Those of skill in the art will readily recognize a variety of noncritical parameters which can be changed or modified to yield essentially the same resultsExample 1Identifying Bacterial Culture Conditions

[0113] Optimized the growth conditions such as medium, pH, and temperature for culturing sulfur-oxidizing bacteria (SOB) for six different SOB species are shown in Table 1.Table 1 - Bacteria Tested

[0114] Each of the bacteria were tested for growth in Thiosulfate broth, NCIM broth, Starkey broth, DSMZ A. ferroxidance broth, DSMZ, and Nutrient broth. Thiosulfate broth and Nutrient broth provided the best overall growth medium.

[0115] Selecting the best medium for growth of SOB cultures included individually inoculating sterile 2 mL of each medium with each bacteria. The inoculated medium was incubated at room temperature on an arbitrary shaker at 100 rpm. Spectrophotometer readings (optical density) were taken on the Oth, 5th and 10th day after inoculation (DAI) at 600 nm wavelength. On the 10th DAI, pour plating was performed and plates were incubated at 30 °C. After incubation, the number of colony forming units (CFU) were determined and expressed as CFU per mF.

[0116] Each of the bacteria, except Acidithiobacillus ferroxidans, were further tested for growth on Nutrient Agar or Thiosulfate Agar, growth in Thiosulfate broth at a pH of 5, 7, and 9, and growth in Thiosulfate broth at various temperatures 30, 35, 40, and 45° C. Sulfur oxidation efficiency was also determined. The results, in order of the best performing to the worst performing, are shown in Table 2. Best performing was the bacteria with the fastest growth or the higher number of viable cells under the test conditions.

[0117] The pH tolerance assay used Thiosulfate broth medium with pH levels adjusted to 5, 7, and 9 using 0.1 N HCE and 0.1 N NaOH. Ten mL of the medium with different pH levelswere transferred to test tubes and sterilized at 121°C for 15 minutes. One mL of broth culture of SOB was inoculated into the ten mL medium in the sterilized test tubes. The inoculated thiosulphate mediums with different pH were incubated at 30 °C. An uninoculated control was also tested. Two replications for each isolate were tested. Spectrophotometer readings were taken on the Oth, 5th, and 10th DAI at 600 nm. It was determined that alkaline pH supports overall good growth of SOB.

[0118] The pH Reduction assay used Thiosulfate broth medium with pH adjusted to 8 using 0.1 N NaOH. A few drops of Bromothymol blue were added to the medium as a pH indicator. Twenty mL of the medium was transferred to 100 mL conical flasks and sterilized at 121 °C for 15 minutes. Two mL of broth cultures of SOB were inoculated to all sterilized flasks and incubated at 30 °C. An uninoculated control was also tested. Two replications for each isolate were tested. Reduction in the pH of the medium was monitored using a digital pH meter and pH strips as well as by observing the change in the color of the broth from blue to yellow. Thiobacillus novellus significantly reduced the pH of the broth.

[0119] The Temperature Tolerance assay tested the effect of various temperatures, 30, 35, 40, and 45 °C, on the growth of the sulfur-oxidizing bacterial cultures. Around 250 mL of thiosulfate broth medium was prepared and 20 mL of the medium was transferred to each of thirteen 100 mL conical flasks. The flasks were sterilized in an autoclave at 121°C for 15 min at 15 psi. Two mL of each SOB broth culture was inoculated into two conical flasks and incubated at 30 °C in an incubator. The same procedure was repeated for the remaining three temperatures (35, 40, 45°C). T. novellus tolerated the highest temperature.

[0120] The S° Oxidation Efficiency assay used modified Thiosulfate broth medium where the thiosulfate was replaced with elemental S (S°). S° served as a source of energy and the growth of the cultures were directly proportional to the oxidation of S°. 50 mL of the medium was transferred to 250 mL conical flasks and sterilized at 121°C for 15 minutes. One mL of broth cultures of SOB was inoculated to all sterilized test tubes and incubated at 30 °C. An uninoculated control was also tested. Three replications for each isolate were maintained. The CFU per mL was observed on the Oth DAI and at 10 DAI by a serial dilution and plating technique. The results, in order of the best performing to the worst performing, are shown in Table 2.Among the tested bacteria, Thiobacillus novellus is the best performing SOB.Table 2 - Order of Best Performance for Bacteria TestedExample 2Bacterium Survival Studies on A Solid Fertilizer

[0121] Thiobacillus novell s was combined with talc (100% Talc) or 75 % tac and 25% elemental sulfur (Talc + S) to create a coating material and coated on urea or glass beads. The coating material and the coated urea was tested for colony forming units (CFUs) at 0, 30, 60, 90, 120, 150, or 180 days after coating the urea or glass beads (DAC).

[0122] Briefly, tissue culture bottles, petri plates, conical flasks, glass rods, screwcap tubes, micropipette tips, test tubes, glass spreaders, and other required lab equipment were sterilized three times in an autoclave at 121° C for 15 minutes at 15 pounds per square inch (psi). Urea to be coated was sterilized by heating to 65° C for three days, followed by exposure to UV rays for 1 hour.

[0123] Thiobacillus novell s was grown in Nutrient broth supplemented with 2% polyvinyl pyrrolidone, used as a cell protectant, and 0.2% potassium sorbate, used as a preservative. After growth reached the desired level, the broth was centrifuged at 10,000 revolutions per minute (rpm) for 10 minutes. The pellet was collected in 15 mL of phosphate buffer supplemented with 1% glycerol in sterilized screw cap tubes.

[0124] The Thiobacillus novellas was added to the 100% Talc coating material or the Talc + S coating material. The coating materials were then tested or stored for further testing orcoated on the urea or glass beads. The coated urea and coated glass beads were then tested or stored for further testing.

[0125] Samples were diluted by taking 1 g of sample from the coating materials (Sample # 1 and 2) and combining with 10 mL water, then serially diluted to 107, 108, 109, and IO10final dilutions or taking 5 g of sample from the coated urea or coated glass beads (Sample # 3, 4, and 5) and combining with 100 mL water, then serially diluted to 103, 104, 105, and 106final dilutions. From each of the final dilutions 0.1 mL was transferred to petri plates containing solidified Nutrient agar medium and spread using a sterile glass spreader. The plates were then incubated at 30° C for 48 hours, after which the number of colonies formed were counted.

[0126] The results of the testing are shown in Table 3. The results demonstrate that the coated bacteria can survive for at least 150 days even when coated on urea and in the presence of elemental sulfur.Table 3 - CFUs per gram of sample (CFU / g)Example 3Changes to Plant Growth

[0127] Thiobacillus novellus was combined with talc (100% Talc) or 75 % tac and 25% elemental sulfur (75% Talc + S) or 50 % tac and 50% elemental sulfur (50% Talc + S) to create a coating material and coated on urea or glass beads using the same methods described in Example 2. The coating material and the coated urea were tested for changes in plant height, stem girth, and biovolume index volume 45 days after addition of 0.5 g of the fertilizer into 5kg soil of soybean plants in pots. Each of the pots were also treated with 100% Recommended Dose of Fertilizer (RDF), and 100% RDF only treated pots were used as a negative control. The RDF (N:P:K 16:32: 15 per acre) per pot was calculated based on the number of plants per acre and applied in two doses (first dose during sowing and the second dose during flowering stage). Plants were watered regularly and maintained till harvest (90-100 days).

[0128] The results of the testing are shown in Table 4. Results that are significantly different from each other are indicated by a matching superscript letter. The treatment using 75% talc and 25% elemental sulfur (75% Talc + S) significantly improved plant growth.Table 4 - Changes to Plant GrowthExample 4Shelf- life Studies on Microbial Formulations

[0129] Thiobacillus novellus was combined with 75 % tac and 25% elemental sulfur (75% Talc + S) or 75 % biochar and 25% elemental sulfur (75% biochar + S) or 75 % bentonite and 25% elemental sulfur (75% Bentonite + S) to create a coating material. The coating materials were coated on wax coated urea or coated on granular elemental S granules (S°) using the same methods described in Example 2.

[0130] The coating materials (T1-T3), the coated wax coated urea formulations (T4-T6), and the coated S° granule formulations (T7 -T9) were tested for colony forming units (CFUs) at 0, 30, 60, 90, 120, 150, or 180 days after coating (DAC).

[0131] The results of the testing are shown in Table 5. T1-T3 treatment test results showed that talc and biochar are superior carriers as compared to bentonite / clay in terms of protecting microbes during long-term storage (IO10vs 107at 180 DAC). Similar to T1-T3 results, T4-T6 treatment test results displayed talc and biochar are superior carriers as compared to Bentonite clay in terms of protecting microbes during long-term storage (106vs 105at 180 DAC). For the coated S° granule formulations, T7-T9 treatment test results, all carriers such as talc, biochar or bentonite look similar, but a greater reduction from the starting CFU counts is seen for use of bentonite. Starting populations on the coated fertilizers were less (109) than T1-T3 (IO12-13) due to dilution with a carrier material.Table 5 - CFUs per gram of sample (CFU / g)Example 5Stability Studies of Microbial Formulations

[0132] The survival of Thiobacillus novellus in a liquid inoculant form was tested when it was directly coated on urea fertilizer. The results indicate that Thiobacillus novellus maintained stable populations (>107CFU / mL) over 5 days in liquid inoculant form (control), while viability declined (<107CFU / mL) when directly coated on urea. The results of the testing are shown in Table 6 and FIG. 3 A and FIG. 3B. This suggests that direct contact with urea compromises Thiobacillus survival, possibly due to chemical stress or osmotic imbalance. Table 6:

Claims

CLAIMSWe claim:

1. A fertilizer composition comprising: a plurality of one or more live bacteria of a Starkeya genus and / or a Thibacillus genus; talc; elemental sulfur; and a solid fertilizer comprising nitrogen, phosphorous, and / or potassium.

2. The fertilizer composition of claim 1, wherein the bacteria comprises Starkeya novella, Acidithiobacillus ferroxidans , Cytobacillus firmus, Psedomonas stutzeri, and / or Enter obacter liiduigii, preferably Starkeya novella.

3. The fertilizer composition of any one of claims 1 to 2, wherein the elemental sulfur comprises micronized sulfur.

4. The fertilizer composition of any one of claims 1 to 3, wherein at least a portion of the bacteria, the talc, and / or the elemental sulfur are comprised in a coating on the solid fertilizer.

5. The fertilizer composition of any one of claims 1 to 4, wherein the composition further comprises a water-soluble adhesive agent, preferably glycerol, carboxy methyl cellulose (CMC), polyvinyl pyrrolidone (PVP), gum arabic, guar gum, monosaccharides of CMC, monosaccharides of arabic gum, monosaccharides of guar gum, disaccharides of CMC, disaccharides of arabic gum, disaccharides of guar gum, or combinations thereof.

6. The fertilizer composition of claim 5, wherein the composition comprises 0.5-2 wt.% of the water-soluble adhesive agent.

7. The fertilizer composition of any one of claims 1 to 6, wherein the composition further comprises: secondary nutrients comprising calcium and / or magnesium; trace elements comprising boron and / or zinc;. a stabilizer comprises starch, agar, polyethylene glycol, polyvinyl alcohol, ethanol, or combinations thereof; and / or a carrier comprising biochar and / or a polysaccharide.

8. The fertilizer composition of any one of claims 1 to 7, wherein the composition comprises 90 to 98 wt.% of the solid fertilizer, 1 to 8 wt.% of the talc, 1 to 3 wt.% of the elemental sulfur, and / or 0.05 to 0.5 wt. % of the plurality of bacteria based on the total weight of the fertilizer composition.

9. The fertilizer composition of any one of claims 1 to 8, wherein at least a portion of the bacteria are at least partially surrounded by the elemental sulfur and / or the talc, and / or wherein at least a portion of the solid fertilizer is at least partially coated by the bacteria, the elemental sulfur, and / or the talc.

10. The fertilizer composition of any one of claims 1 to 9, wherein the fertilizer is homogenous.

11. A method of making the fertilizer composition of any one of claims 1 to 9, the method comprising coating an outer surface of a core comprising the solid fertilizer with the elemental sulfur, the talc, and / or the bacteria, preferably contacting the outer surface of the core with a solution comprising the elemental sulfur, the talc, and / or the bacteria.

12. The method of claim 11, wherein the coating is formed by contacting the outer surface of the core with a water-soluble adhesive agent prior to contact with the elemental sulfur, the talc, and / or the bacteria.

13. The method of any one of claims 11 to 12, further comprising combining the bacteria and the talc before contacting the bacteria and the talc with the fertilizer.

14. A method of making the fertilizer composition of claim 10, the method comprising combining and mixing the solid fertilizer with the elemental sulfur, the talc, and / or the bacteria to form a homogenous composition.

15. A method of fertilizing, the method comprising applying the fertilizer composition of any one of claims 1 to 10 to a soil, a crop, water, or any combination thereof.

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