A soil treatment agent and manufacturing method thereof

Abstract

The present invention pertains to a soil treatment agent and its manufacturing method. The manufacturing method for the soil treatment agent comprises the steps: S101 : mixing a raw soil material with a cultivation medium to achieve a first mixture; wherein the raw soil material comprises original microorganisms that are viable; S102: cultivating the first mixture for a first cultivation period under a first cultivation condition to achieve a second mixture, wherein the first cultivation condition is favourable for promoting growth of the original microorganisms; S103: assessing viable microorganism concentration of the second mixture to determine whether the viable microorganism concentration of the second mixture has achieved a predetermined training threshold; S104: if the predetermined training threshold is achieved, then subjecting the second mixture to a treatment protocol comprising a plurality of treatment cycles to achieve the soil treatment agent, wherein each treatment cycle comprises temperature-shock treatment and oxygen-shock treatment.

Description

DESCRIPTIONTITLE OF INVENTION: [A SOIL TREATMENT AGENT AND MANUFACTURING METHOD THEREOF]FIELD

[0001] The present invention relates to the field of soil treatment or soil remediation, and particularly to a soil treatment agent and manufacturing method thereof.BACKGROUND ART

[0002] The following discussion of the background to the disclosure is intended to facilitate an understanding of the present disclosure only. It should be appreciated that the discussion is not an acknowledgement or admission that any of the material referred to was published, known or part of the common general knowledge of the person skilled in the art in any jurisdiction as at the priority date of the disclosure.

[0003] Black soil," also known as "chernozem," represents a type of soil abundant in humus and is highly favorable for agricultural cultivation. Enriched with minerals such as phosphoric acid, natural humic substance (distinct from mere mineral-source humic acid), phosphorus, ammonia, magnesium, calcium, and iron, it exhibits a substantial concentration of natural humic substance, ranging between 20% to 25%. This unique composition contributes to stabilizing the soil's microbial ecology. Consequently, it helps in maintaining the soil's physical structure, chemical properties, and biological ecosystem with consistency. Unlike other soil types, black soil can meet the agricultural planting requirements effectively without the dependency on chemical fertilizers, reflecting its innate capacity to support sustainable farming practices.

[0004] Black soils have many advantages over the commonly used chemical fertilizers. The common practice of over-cultivation and overreliance on chemical fertilizers and pesticides has had detrimental effects, leading to imbalances in soil pH, salinization, compaction, and the near- total eradication of microbial diversity. These disruptions to the soil's natural equilibrium contribute to a loss of fertility, rendering farming more challenging and less productive, with reduced yields, increased reliance on fertilizers, and escalatedprevalence of soil-bome diseases. Moreover, the predominant use of chemical fertilizers in contemporary farming — with no regard for the maturity or natural humic substance content in current organic fertilizer standards — exacerbates these problems. Some practices even include the addition of mineral- source humic acid extracted through coal industry acid washing. This widespread misuse of chemical fertilizers disrupts the soil's natural balance further, depleting its vitality, and effectively contributing to environmental pollution, highlighting an urgent need for responsible and sustainable soil management practices. Thus, there is a growing interest to exploit the use of black soil.

[0005] However, existing reserves of black soil are extremely limited, and arbitrary extraction of black soil is prohibited by regulations in many countries. Accordingly, efforts have made to create the so-called “artificial black soil”. For example, US patent application document US20230217872A1, titled “Preparation Method for Artificially Simulated Soil Rich in Humus,” describes a method for creating artificial soil that's rich in humus and is claimed to resemble natural "black soil." This method includes preparing a soil mixture and then subjecting it to a hydrothermal reaction in a high-temperature, high-pressure kettle (e.g., 180°C - 200°C under a pressure of 0.5MPa - 5MPa for 20 - 28 hours). The result is an artificially simulated soil that's said to be rich in humus. Despite these claims of fertility, the artificial soil might fall short in replicating the various attributes of natural black soil, including its healthy and robust micro-ecosystem. The current method's ability to imitate the complex properties that define natural black soil is uncertain, suggesting potential limitations in its effectiveness.

[0006] Other methods, such as those described in the Chinese patent application document CN113182325A, titled “Method for preparing artificial black soil from household kitchen garbage,” purport to create artificial black soil by introducing “high activity facultative anaerobic microbial floras” into household kitchen garbage for fermentation. However, the efficacy of using only these specific microbial floras to transform kitchen waste into a substance akin to black soil is questionable. It is well understood by those skilled in the field that black soil hosts a complex and diverse range of microorganisms that together create a healthy and robust micro-ecosystem.

[0007] The present disclosure contemplates that it would be desirous to provide a soil treatment agent and manufacturing method thereof, which may at least alleviate or mitigate the afore-mentioned technical problems.SUMMARY

[0008] In view of the above technical problems, the present invention provides a soil treatment agent and manufacturing method thereof that might partially solve or alleviate the above-mentioned deficiencies in the prior art and can provide a more efficient and effective technical solution for improving the soil properties.

[0009] Specifically, to address the above-mentioned technical problems, the present invention specifically uses the following technical solutions:

[0010] In accordance with one aspect of the present invention, there is provided a manufacturing method for a soil treatment agent, wherein comprising following steps:

[0011] S 101 : mixing a raw soil material with a cultivation medium to achieve a first mixture; wherein the raw soil material comprises original microorganisms that are viable.

[0012] S102: cultivating the first mixture for a first cultivation period under a first cultivation condition to achieve a second mixture, wherein the first cultivation condition is favorable for promoting growth of the original microorganisms.

[0013] S103: assessing viable microorganism concentration of the second mixture to determine whether the viable microorganism concentration of the second mixture has achieved a predetermined training threshold.

[0014] S104: if the predetermined training threshold is achieved, then subjecting the second mixture to a treatment protocol comprising a plurality of treatment cycles to achieve the soil treatment agent, wherein each treatment cycle comprises temperature- shock treatment and oxygen- shock treatment.

[0015] In some embodiments, the raw soil material is selected from the group consisting of black soil, peat soil, brown forest soil, artificially modified or treated soils rich in beneficial microorganisms, and any combination thereof.

[0016] In some embodiments, the cultivation medium comprises a mixture of lysogeny broth powder, agar, yeast extract, trace composite amino acids and water.

[0017] In some embodiments, the manufacturing method further comprises S105: if the viable microorganism concentration of the second mixture has yet to achieve the predetermined training threshold, cultivating the second mixture further for a second cultivation period under a second cultivation condition, wherein the second cultivation condition is more favorable than the first cultivation condition for promoting the growth of the original microorganisms.

[0018] In some embodiments, the predetermined training threshold is deemed achieved if a viable cell count of the second mixture is at least 1.0 x 108CFU / g.

[0019] In some embodiments, the temperature-shock treatment comprises a low- temperature training session and a high-temperature training session; wherein the low- temperature training session involves placing the second mixture in a low-temperature environment of 0°C - 10°C, and the high-temperature training session involves placing the second mixture in a high-temperature environment of 50°C - 85°C; wherein each training session of the temperature- shock treatment is arranged to last 7 - 10 days.

[0020] In some embodiments, the oxygen- shock treatment comprises a low-oxygen training session and a high-oxygen training session; wherein the low-oxygen training session involves subjecting the second mixture to a low-oxygen environment with dissolved oxygen concentration between 0.1 - 0.4 mg / L; and the high-oxygen training session involves subjecting the second mixture to a high-oxygen environment with dissolved oxygen concentration between 4.0 ~ 6.0mg / L; wherein each training session of the oxygen-shock treatment is arranged to last 3 - 5 days. Different techniques can be employed to administer the oxygen-shock. For instance, in certain embodiments, a method involving intermittent aeration is favored. This approach uses an oxygen feed pump equipped with a precision pressure-regulating throttle valve, allowing for dynamic adjustment and stabilization of the dissolved oxygen concentration within the second mixture.

[0021] In some embodiments, the manufacturing method further comprises S106: monitoring the viable microorganism concentration of the second mixture to determine whether the viable microorganism concentration has been maintained at or above a viability threshold during operation of the treatment protocol.

[0022] In some embodiments, if the viable microorganism concentration of the second mixture has dropped below the viability threshold, temporarily pausing the treatment protocol, and subjecting the second mixture to a recovery environment for a recovery period till the viable microorganism concentration has increased to or above the viability threshold.

[0023] In some embodiments, the viability threshold is deemed maintained if the viable microorganism count of the second mixture is at least 1.0 x 106CFU / g.

[0024] In some embodiments, temperature- shock treatment and oxygen- shock treatment are arranged to take place separately in sequence.

[0025] In some embodiments, in one or more of the treatment cycles, the temperatureshock treatment and the oxygen-shock treatment are arranged to be conducted simultaneously; wherein the temperature- shock treatment comprises a low-temperature training session and a high-temperature training session; and the oxygen-shock treatment comprises a low-oxygen training session and a high-oxygen training session; wherein the low-temperature training session and the low-oxygen training session are arranged to be conducted simultaneously; and the high-temperature training session and the high-oxygen training session are arranged to be conducted simultaneously.

[0026] In accordance with another aspect of the present invention, there is a provided a soil treatment agent manufactured according to the manufacturing methods described above, wherein a viable cell count of the soil treatment agent is at least 1.5xl08CFU / g.

[0027] In accordance with another aspect of the present invention, there is a provided a manufacturing method for a soil treatment composition, comprising following steps:

[0028] S107: introducing the soil treatment agent of Claim 13 into a compost base material to achieve a weight percentage of the soil treatment agent over the compost basematerial between 0.3% to 0.5%; and S108: cultivating the compost base material under a third cultivation condition for a third cultivation period.

[0029] In some embodiments, the compost base material is selected from the group consisting of discarded mushroom cultivation substrate, livestock and poultry manure, agricultural waste, and any combination thereof.

[0030] In some embodiments, the compost base material is prepared to have a carbon- to-nitrogen ratio between 15 and 25, a moisture content between 50% and 60%, and a pH value between 6 and 9.

[0031] In some embodiments, the third cultivation condition is arranged to have a temperature between 22°C - 28°C.

[0032] In some embodiments, the third cultivation period is at least 30 days, optionally 35 - 45 days.

[0033] In some embodiments, the manufacturing method further comprises S109: monitoring contents of (i) humic substance; (ii) organic substance; (iii) total content of nitrogen, phosphorus, and potassium; and (iv) live microorganisms, in the soil treatment composition prepared from S108.

[0034] In some embodiments, the manufacturing method further comprises SI 10: harvesting the soil treatment composition once (i) the humic substance is assessed to have reached a humic substance concentration of at least 18% by weight over the soil treatment composition; (ii) the organic substance is assessed to have reached an organic substance concentration of at least 80% by weight over the soil treatment composition; (iii) the total content of nitrogen, phosphorus, and potassium is assessed to have reached a NPK concentration of at least 7%; and (iv) viable cell count of the live microorganisms is assessed to have reached at least 1.5xl09CFU / g.

[0035] In summary, the manufacturing methods of the present invention utilize a minimal amount of soil raw material to develop a soil treatment agent that effectively preserves the beneficial microorganisms originally inhabiting the soil. Through a carefully orchestrated treatment protocol that includes shock treatments (such as temperature -shock and oxygen-shock treatments), the method imposes selective stresses on the microorganismsduring the development of the soil treatment agent. This ensures that microorganisms adapted to endure harsh environmental conditions are not outcompeted, allowing their unique features and capabilities to be preserved through successive generations of propagation. These attributes are vital for fostering a healthy and resilient micro-ecosystem when the soil treatment agent as well as the soil treatment composition developed (via fermentation) using the soil treatment agent is applied to rehabilitate degraded soils or to cultivate fertile lands akin to specific soil types such as black soil, peat soil, or brown forest soil.

[0036] In addition, the manufacturing methods provided in the present invention neither demand precise identification of the various microorganisms originally found within the soil raw material nor sacrifice the complex interplay among these microorganisms. Instead, they manage to preserve this delicate balance, which is essential in replicating the unique characteristics of natural black soil. This approach recognizes the inherent complexity of the soil ecosystem and leverages it, rather than trying to isolate or artificially recreate specific components.

[0037] Other aspects of the disclosure will be apparent to those of ordinary skill in the art upon review of the following description of specific embodiments of the disclosure in conjunction with the accompanying drawings.BRIEF DESCRIPTION OF THE DRAWINGS

[0038] In the figures, which illustrate, by way of non-limiting examples only, embodiments of the present invention.

[0039] Fig. 1 is a flowchart illustrating some of the steps of the manufacturing method for the soil treatment agent of the present invention.

[0040] Fig. 2 is a flowchart illustrating some further steps of the manufacturing method for the soil treatment agent of the present invention.

[0041] Fig. 3 is a flowchart illustrating an example of the treatment protocol.

[0042] Fig. 4 is a flowchart illustrating an example of the treatment cycles.

[0043] Fig. 5 is a flowchart illustrating some of the steps of the manufacturingmethod for the soil treatment composition of the present invention.

[0044] Corresponding parts are denoted by the same reference numerals from Figures 1 to 5.DETAILED DESCRIPTION

[0045] In the following description, details are provided to describe the embodiments of the specification. It shall be apparent to one skilled in the art, however, that the embodiments may be practiced without such details.

[0046] Similar parts of the embodiments may have the same names, similar part numbers, or similar part numbers with an alphabet symbol or prime symbol. The description of one part applies by reference to another similar part, where appropriate, thereby reducing repetition of text without limiting the disclosure.

[0047] Throughout this specification, unless otherwise indicated to the contrary, the terms ‘comprising’, ‘consisting of, ‘having’ and the like, are to be construed as non- exhaustive, or in other words, as meaning ‘including, but not limited to’.

[0048] Throughout the specification, unless the context requires otherwise, the word ‘include’ or variations such as ‘includes’ or ‘including’ will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers.

[0049] Throughout the specification, the singular form "a", "an" and "the" include plural references unless the context clearly dictates otherwise. For example, the term "a bioreactor" or "at least one bioreactor" may include a plurality of bioreactors.

[0050] Throughout the specification, the term “black soil”, also known as “chernozem”, refers to a black-colored soil containing a high percentage of humus (4% to 16%) and high percentages of phosphorus and ammonia compounds. Black soil is known to be very fertile soil and can produce high agricultural yields with its unique properties, such as high moisture storage capacity.

[0051] Throughout the specification, the term “humic substance” refers to decomposed organic matter, more specifically coloured recalcitrant organic compounds that might be naturally or artificially formed during long-term decomposition and transformation of biomass residues. The color of humic substances varies from yellow to brown to black. Humic substance include humic acid, fulvic acid, humin and hymatomelanic acid which differ in solubility.

[0052] Throughout the specification, the term “beneficial microorganism” encompasses bacteria, archaea, fungi, and protozoa that contribute positively to the soil's ecosystem. These microorganisms foster a healthy and robust micro-ecosystem within the soil, enhancing advantageous properties such as moisture retention capacity. Furthermore, they play a role in inhibiting the growth of plant-pathogenic microorganisms, thus maintaining, or even improving soil quality and functionality.

[0053] Throughout the specification, the term “viable cell count” refers to the colony forming units (“CFU”) per gram of soil, e.g., CFU / g, of a microorganism as measured by, for example, the standard dilution plating methods. In the context of the present invention, measuring the viable cell count is one of the possible methods to evaluate the microorganism's overall viability during the development of the soil treatment agent and the soil treatment composition. This assessment serves as a guide to determine whether the manufacturing process can advance to the next phase, such as commencing the treatment protocol once the microorganisms reach the training threshold. Conversely, it can also signal a need to pause the process, for example, if a training session imposes excessive stress on the microorganisms originally present in the soil raw material, thereby necessitating a temporary halt for recovery. Importantly, the viable cell count is not an exhaustive indicator of the types and quantities of microorganisms in the soil raw material or the manufactured soil treatment agent. A skilled practitioner will recognize that these may include microorganisms that are not detectable using conventional techniques like plating methods, thus requiring careful interpretation and consideration in the process.

[0054] Throughout the specification, the term “compost base material” refers to the foundational material used in the composting process, typically consisting of organic matter that will break down into compost over time.

[0055] Unless defined otherwise, all other technical and scientific terms used herein have the same meaning as is commonly understood by a skilled person to which the subject matter herein belongs.

[0056] Embodiment.1 A Manufacturing Method for A Soil Treatment Agent

[0057] Referring to Fig. 1, the present invention provides a manufacturing method for a soil treatment agent, wherein comprising following steps:

[0058] S101: mixing a raw soil material with a cultivation medium to achieve a first mixture; wherein the raw soil material comprises original microorganisms that are viable.

[0059] In some embodiments, the raw soil material is selected from the group consisting of black soil, peat soil, brown forest soil, artificially modified or treated soils rich in beneficial microorganisms, and any combination thereof.

[0060] In some embodiments, the cultivation medium comprises a composition of lysogeny broth powder, agar, yeast extract, trace composite amino acids and water. The aim of this composition is to create a cultivation medium tailored to promote the growth of a diverse range of microorganisms.

[0061] The objective of this step is to utilize a soil source, either known or potentially rich in beneficial microorganisms, as a foundational material. By capturing and cultivating these microorganisms, a soil treatment agent can be developed to improve soil quality. This method obviates the need to identify the specific types of beneficial microorganisms in the soil source, thereby circumventing the usual technical challenges of isolating and growing unknown or undetectable microorganisms. However, it remains crucial to ensure that the microorganisms in the soil source are still viable. To this end, using freshly prepared soil or soil that has been properly stored in a controlled environment (e.g., with regulated humidity, temperature, and air) is essential for the success of the method described in the present invention.

[0062] S102: cultivating the first mixture for a first cultivation period under a first cultivation condition to achieve a second mixture, wherein the first cultivation condition isdesigned to foster the growth of the original microorganisms within the soil source. The goal of this step is to create an environment conducive to the rapid expansion of the diverse microorganisms naturally present in the soil, enhancing their propagation.

[0063] S103: assessing viable microorganism concentration of the second mixture to determine whether the viable microorganism concentration of the second mixture has achieved a predetermined training threshold. Various methods suitable for determining both viability and concentration of microorganisms may be employed, including, but not limited to, plate counting, flow cytometry, turbidity measurement, respiration measurements, and microscopic examination with staining. It should be understood by those skilled in the field that the assessment of viable microorganism concentration in this context is intended for a general or rough estimation of the overall viability of the diverse microorganisms present. It is not aimed at accurately identifying every individual organism from the soil source, as many types of microorganisms may be undetectable using conventional detection methods.

[0064] The objective of this step is to assess whether the communities of viable microorganisms within the second mixture have reached a concentration level ready for treatment under harsh environmental conditions, known as the training threshold. It is beneficial to halt the expansion of the viable microorganisms once this threshold has been achieved, as excessive growth may present disadvantages. Since the first cultivation condition is favorable to most microorganisms, those more suited to thrive in friendly or comfortable environments might overgrow, outcompeting those adapted to survive under harsh conditions. Furthermore, overexpanding even the microorganisms capable of enduring severe environments might lead to offspring with diminished capabilities and loss of specific features important for creating healthy and robust micro-ecosystem.

[0065] S104: if the predetermined training threshold is achieved, then subjecting the second mixture to a treatment protocol comprising a plurality of treatment cycles to achieve the soil treatment agent, wherein each treatment cycle comprises temperature- shock treatment and oxygen-shock treatment. In some embodiments, the predetermined training threshold is deemed achieved if a viable cell count of the second mixture is at least 1.0 x 108CFU / g. Trialexperiments show that, starting the training with the viable cell count at or above 1.0 x 108CFU / g helps achieve relatively more stable and consistent production of the soil treatment agent rich in live microorganisms that manage to survive the harsh treatments.

[0066] S105: if the viable microorganism concentration of the second mixture has yet to achieve the predetermined training threshold, cultivating the second mixture further for a second cultivation period under a second cultivation condition, wherein the second cultivation condition is more favorable than the first cultivation condition for promoting the growth of the original microorganisms. This step can be particularly useful when encountering challenges in growing certain microorganisms, allowing for the application of an even more accommodating environment to aid their growth. With this approach, the first cultivation condition can be tailored to be moderately conducive to growth, avoiding an overly hospitable environment that might enable organisms suited to thrive in friendly conditions to quickly outcompete those adapted to harsher environments. If the first cultivation condition does not lead to healthy expansion, it can be adjusted to the second cultivation condition to facilitate the growth of the microorganisms. This ensures a balanced approach that promotes growth while still selecting for resilience.

[0067] In some embodiments, the temperature-shock treatment comprises a low- temperature training session and a high-temperature training session; wherein the low- temperature training session involves placing the second mixture in a low-temperature environment of 0°C - 10°C for a first treatment duration (e.g., 7 - 10 days), and the high- temperature training session involves placing the second mixture in a high-temperature environment of 50°C - 85°C for a second treatment duration (e.g., 7 - 10 days).

[0068] In some embodiments, the oxygen- shock treatment comprises a low-oxygen training session and a high-oxygen training session; wherein the low-oxygen training session involves subjecting the second mixture to a low-oxygen environment with dissolved oxygen concentration between 0.1 - 0.4 mg / L for a third treatment duration (e.g., 3 - 5 days); and the high-oxygen training session involves subjecting the second mixture to a high-oxygenenvironment with dissolved oxygen concentration between 4.0 - 6.0 mg / L for a fourth treatment duration (e.g., 3 - 5 days).

[0069] As shown in Table 1, the specified training conditions described above are preferred in certain embodiments, for example, those involving precious and limited raw soil sources / These conditions appear to balance the preservation of native microorganism viability with their exposure to challenging environmental conditions, which helps the offspring inherit and retain vital traits essential for establishing a healthy and resilient microecosystem within the soil.

[0070] Referring to Fig. 2, in some embodiments, the manufacturing method further comprises S106: monitoring the viable microorganism concentration of the second mixture to determine whether the viable microorganism concentration has been maintained at or above a viability threshold during operation of the treatment protocol. In some embodiments, the viability threshold is deemed maintained if the viable microorganism count of the second mixture is at least 1.0 x 106CFU / g.

[0071] The objective of this step is to dynamically monitor the stress caused by the shock treatments to ensure that at least those microorganisms adapted to harsh conditions can still survive. This monitoring helps to calibrate the intensity of the treatments, thereby preventing them from becoming excessively harsh. For example, once it is found that the viable microorganism concentration of the second mixture has dropped below the viability threshold, it is advantageous to halt the treatment protocol, and to subject the second mixture to a recovery environment for a recovery period till the viable microorganism concentration has increased to or above the viability threshold.

[0072] This step enables the treatment protocol to initiate with one or more treatment cycles, characterized by relatively harsh training sessions (e.g., more extreme temperatures or dissolved oxygen concentrations). Should these training sessions lead to a substantial decrease in the concentration of viable microorganisms, the second mixture can then undergo a more relaxed treatment cycle following a recovery period. The primary goal is to strike a balance in applying stresses at the onset of the treatment protocol, thereby efficientlyselecting the intended microorganisms vital for improving soil quality, without losing them through the application of overly harsh or inadequate conditions.

[0073] Referring to Fig. 3, in some embodiments, in one or more of the treatment cycles (201, 203 and 205), temperature- shock treatment and oxygen-shock treatment are conducted separately and in sequence. This approach is particularly useful in scenarios where the microorganisms appear less resilient to harsh environmental conditions. By exposing them to one challenging condition at a time, a more buffered environment is provided, allowing the microorganisms to better endure the stresses.

[0074] Conversely, in other embodiments (referring to Fig. 4), in one or more of the treatment cycles (201, 203), the temperature- shock treatment and oxygen-shock treatment are arranged to be carried out simultaneously. This approach imposes multiple harsh environmental stresses, aiming to select robust microorganisms capable of forming a strong micro-ecosystem to enhance soil quality. For instance, the temperature -shock treatment may include a low-temperature training session and a high-temperature training session; likewise, the oxygen-shock treatment might consist of a low-oxygen training session and a high- oxygen training session. These can be arranged such that the low-temperature session is conducted simultaneously with the low-oxygen session (301, 305), and the high-temperature session is conducted simultaneously with the high-oxygen session (303, 307). This simultaneous treatment ensures facilitates a stringent selection process, favoring the survival of microorganisms resilient to multi-dimensional environmental stressors. Under the simultaneous -treatment arrangement, the duration of each training session, in which two or multiple treatment conditions are being applied, is suggested to be shorten. For example, when subjecting the second mixture to low temperature of 8 °C and low dissolved oxygen of 0.4 mg / L concurrently, the duration of the training session could be limited to 5 days, ensuring a consistent and stable production of a soil treatment agent with viable cell counts of at least 1.5xl08CFU / g.

[0075] In accordance with another aspect of the invention, there is provided a soil treatment agent manufactured according to the manufacturing methods described above. Inspecific embodiments, the harvesting of the soil treatment agent is initiated when the concentration of viable microorganisms within reaches or exceeds a threshold of 1.5xl08CFU / g. This ensures a potent formulation of soil treatment agent rich in microorganisms that can be effectively applied to ferment the compost base material (e.g., discarded mushroom cultivation substrate, livestock and poultry manure, agricultural waste) to obtain the soil treatment composition.

[0076] Embodiment.2 A Manufacturing Method for A Soil Treatment Composition

[0077] Referring to Fig. 5, in accordance with another aspect of the invention, there is provided a manufacturing method for a soil treatment composition, wherein comprising following steps:

[0078] SI 07: introducing the soil treatment agent harvested from Embodiment.1 into a compost base material to achieve a weight percentage of the soil treatment agent over the compost base material between 0.3% to 0.5%. This concentration ensures optimal fermentation powered by the microorganisms that were trained and remained viable in the soil treatment agent. Otherwise, the beneficial microorganisms that underwent training and survived might be outcompeted by less desirable microorganisms naturally present in the compost base material. Such competition can detrimentally influence the fermentation process. For instance, experiments indicate that an insufficient amount of the soil treatment agent can prolong the fermentation, affecting the desired levels of humic substances and organic materials in the soil treatment composition produced in the end. Furthermore, it is evident to a skilled person that starting with a higher concentration of the soil treatment agent, for example, exceeding 0.5% by weight, can offer added benefits and is also a feasible approach.

[0079] S108: cultivating the compost base material under a third cultivation condition for a third cultivation period. This step can employ standard cultivation conditions and periods typically associated with fermentation. Essentially, any condition conducive to thegrowth and proliferation of most microorganisms would be suitable for this step of the present invention.

[0080] In some embodiments, the compost base material is selected from the group consisting of discarded mushroom cultivation substrate, livestock and poultry manure, agricultural waste, and any combination thereof. Beyond the examples cited, other materials traditionally recognized as suitable for composting can also be employed in this step of the invention.

[0081] In some embodiments, the compost base material is prepared to have a carbon- to-nitrogen ratio between 15 and 25, a moisture content between 50% and 60%, and a pH value between 6 and 9. In some embodiments, the third cultivation condition is arranged to have a temperature between 22°C - 28°C. In some embodiments, the third cultivation period is at least 30 days, optionally 35 - 45 days.

[0082] In some embodiments, the manufacturing method further comprises S109: monitoring contents of (i) humic substance (e.g., ); (ii) organic substance; (iii) total content of nitrogen, phosphorus, and potassium; and (iv) live microorganisms, in the soil treatment composition prepared from S108.

[0083] In some embodiments, the manufacturing method further comprises SI 10: harvesting the soil treatment composition once (i) the humic substance is assessed to have reached a humic substance concentration of at least 18% by weight over the soil treatment composition; (ii) the organic substance is assessed to have reached an organic substance concentration of at least 80% by weight over the soil treatment composition; (iii) the total content of Nitrogen, Phosphorus, and Potassium (“NPK”) is assessed to have reached a NPK concentration of at least 7%; and (iv) viable cell count of the live microorganisms is assessed to have reached at least 1.5xl09CFU.

[0084] The table (Table.1) below summarizes experimental results from select embodiments of the present invention. These results highlight the enhanced characteristics of the soil treatment composition produced and harvested using the described methods.Table.1 Soil Treatment Composition Prepared

[0085] In the experiment groups (G1 - G3) shown in the Table.1 above, the raw soil material used is a modest quantity of natural black soil known to be rich in beneficial microorganisms capable of creating a healthy and robust micro-ecosystem. For all three groups, three treatment cycles were conducted, and each treatment cycle encompassed four training sessions: low-temperature (Low-T), high-temperature (High-T), low dissolved oxygen (Low-DO), and high dissolved oxygen (High-DO), which were sequentially executed in isolation. The specifics of each training session's conditions are summarized in Table.1.

[0086] As shown in Table.1, the soil treatment compositions derived from all three groups surpass conventional fertilizers in certain key characteristics, such as the concentration of both humic and organic substances. For context, conventional fertilizers typically contain approximately 30% (w / w) organic substance and around 5% (w / w) humic substance concentration. Notably, the compositions from the three groups of the present invention not only have an increased humic substance content but also a significant portion of that is fulvic acid. Contrastingly, in conventional methods, even those fertilizers boasting high humic substance content predominantly contain humic acid, often with negligible fulvic acid content. Fulvic acids, distinguished by their golden hue, are smaller molecules than humic acids, soluble in water across all pH ranges, and are more biologically active. This is attributed to their oxygen content, which is nearly double that of humic acids, and other advantageous properties, such as being a highly effective carbon-containing chelatingcompound. Therefore, due to their enriched fulvic acid content, the soil treatment compositions of the present invention may, in certain respects, outperform conventional fertilizers that lack fulvic acid.

[0087] Furthermore, among the three groups, G1 with the Low-T between 0°C — 10°C, the High-T between 50°C — 85°C, the Low-DO between 0.1 — 0.4 mg / L and the high-DO between 4.0 — 6.0mg / L outperforms the other two groups (G2 and G3) by achieving the soil treatment composition with notably enhanced levels of humic substances (18 - 22% (w / w)), fulvic acid (9 - 12% (w / w)), and organic materials (more than 80% (w / w)).

[0088] Moreover, the method of G1 demonstrates enhanced consistency and stability in the production of the soil treatment agent, consistently achieving a high concentration of beneficial microorganisms (e.g., at least 1.5xl08CFU / g viable cell count), as well as in the production of the soil treatment composition (i.e., the compost base material fermented by the soil treatment agent), which consistently achieves a high concentration of beneficial microorganisms (e.g., at least 1.5xl09CFU / g viable cell count). In contrast, the methods of G2 and G3, which adopt more aggressive treatment conditions, exhibit relatively less reliability in this aspect. Therefore, in situations where preserving the indigenous microorganisms in valuable and scarce raw soil materials is important, the milder treatment conditions of G1 are then preferred, which could help avoid the unexpected and unintended eradication of native microorganisms that might occur with more aggressive treatment conditions.

[0089] It is to be appreciated by the person skilled in the art that variations and combinations of features described above, not being alternatives or substitutes, may be combined to form yet further embodiments falling within the intended scope of the disclosure.

Claims

CLAIMS1. A manufacturing method for a soil treatment agent, comprising following steps:S 101: mixing a raw soil material with a cultivation medium to achieve a first mixture; wherein the raw soil material comprises original microorganisms that are viable;S102: cultivating the first mixture for a first cultivation period under a first cultivation condition to achieve a second mixture, wherein the first cultivation condition is favorable for promoting growth of the original microorganisms;S103: assessing viable microorganism concentration of the second mixture to determine whether the viable microorganism concentration of the second mixture has achieved a predetermined training threshold;S104: if the predetermined training threshold is achieved, then subjecting the second mixture to a treatment protocol comprising a plurality of treatment cycles to achieve the soil treatment agent, wherein each treatment cycle comprises temperature- shock treatment and oxygen- shock treatment.

2. The manufacturing method according to Claim 1, wherein the raw soil material is selected from the group consisting of black soil, peat soil, brown forest soil, artificially modified or treated soils rich in beneficial microorganisms, and any combination thereof.

3. The manufacturing method according to Claim 1, wherein the cultivation medium comprises a mixture of lysogeny broth powder, agar, yeast extract, trace composite amino acids and water.

4. The manufacturing method according to Claim 1, wherein further comprising S105: if the viable microorganism concentration of the second mixture has yet to achieve the predetermined training threshold, cultivating the second mixture further for a second cultivation period under a second cultivation condition, wherein the second cultivation condition is more favorable than the first cultivation condition for promoting the growth of the original microorganisms.

5. The manufacturing method according to Claim 1, wherein the predetermined training threshold is deemed achieved if a viable cell count of the second mixture is at least 1.0 x 108CFU / g.

6. The manufacturing method according to Claim 1, wherein the temperature- shock treatment comprises a low-temperature training session and a high-temperature training session; wherein the low -temperature training session involves placing the second mixture in a low-temperature environment of 0°C - 10°C, and the high-temperature training session involves placing the second mixture in a high-temperature environment of 50°C - 85°C; wherein each training session of the temperature- shock treatment is arranged to last 7 - 10 days.

7. The manufacturing method according to Claim 1, wherein the oxygen- shock treatment comprises a low-oxygen training session and a high-oxygen training session; wherein the low-oxygen training session involves subjecting the second mixture to a low-oxygen environment with dissolved oxygen concentration between 0.1 - 0.4 mg / L; and the high- oxygen training session involves subjecting the second mixture to a high-oxygen environment with dissolved oxygen concentration between 4.0 ~ 6.0mg / L; wherein each training session of the oxygen-shock treatment is arranged to last 3 - 5 days.

8. The manufacturing method according to Claim 1, wherein further comprising S106: monitoring the viable microorganism concentration of the second mixture to determine whether the viable microorganism concentration has been maintained at or above a viability threshold during operation of the treatment protocol.

9. The manufacturing method according to Claim 8, wherein, if the viable microorganism concentration of the second mixture has dropped below the viability threshold, temporarily pausing the treatment protocol, and subjecting the second mixture to a recovery environment for a recovery period till the viable microorganism concentration has increased to or above the viability threshold.

10. The manufacturing method according to Claim 8 or 9, wherein the viability threshold is deemed maintained if the viable microorganism count of the second mixture is at least 1.0 x 106CFU / g.

11. The manufacturing method according to Claim 1, wherein temperature- shock treatment and oxygen-shock treatment are arranged to take place separately in sequence.

12. The manufacturing method according to Claim 1, wherein, in one or more of the treatment cycles, the temperature- shock treatment and the oxygen- shock treatment are arranged to be conducted simultaneously; wherein the temperature- shock treatment comprises a low-temperature training session and a high-temperature training session; and the oxygen-shock treatment comprises a low-oxygen training session and a high-oxygen training session; wherein the low-temperature training session and the low-oxygen training session are arranged to be conducted simultaneously; and the high-temperature training session and the high-oxygen training session are arranged to be conducted simultaneously.

13. A soil treatment agent manufactured according to anyone of Claims 1 - 12, wherein a viable cell count of the soil treatment agent is at least 1.5xl08CFU / g.

14. A manufacturing method for a soil treatment composition, comprising following steps: S107: introducing the soil treatment agent of Claim 13 into a compost base material to achieve a weight percentage of the soil treatment agent over the compost base material between 0.3% to 0.5%; andS108: cultivating the compost base material under a third cultivation condition for a third cultivation period.

15. The manufacturing method according to Claim 14, wherein the compost base material is selected from the group consisting of discarded mushroom cultivation substrate, livestock and poultry manure, agricultural waste, and any combination thereof.

16. The manufacturing method according to Claim 14, wherein the compost base material is prepared to have a carbon-to-nitrogen ratio between 15 and 25, a moisture content between 50% and 60%, and a pH value between 6 and 9.

17. The manufacturing method according to Claim 14, wherein the third cultivation condition is arranged to have a temperature between 22°C - 28°C.

18. The manufacturing method according to Claim 14, wherein the third cultivation period is at least 30 days, optionally 35 - 45 days.

19. The manufacturing method according to Claim 14, wherein further comprising S109: monitoring contents of (i) humic acid; (ii) organic substance; (iii) total content of nitrogen, phosphorus, and potassium; and (iv) live microorganisms, in the soil treatment composition prepared from S108.

20. The manufacturing method according to Claim 19, wherein further comprising SI 10: harvesting the soil treatment composition once (i) the humic acid is assessed to have reached a humic acid concentration of at least 18% by weight over the soil treatment composition; (ii) the organic substance is assessed to have reached an organic substance concentration of at least 80% by weight over the soil treatment composition; (iii) the total content of nitrogen, phosphorus, and potassium is assessed to have reached a NPK concentration of at least 7%; and (iv) viable cell count of the live microorganisms is assessed to have reached at least 1.5xl09CFU / g.

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