Preparation method and application of culture medium filler

Abstract

The invention provides a preparation method and application of a culture medium filler. The preparation method comprises the following steps: S1) sequentially soaking an initial culture medium filler in a first nutrient solution and a second nutrient solution to obtain a first culture medium filler; s2) performing first mixing on the first culture medium filler and a mushroom liquid to obtain a second culture medium filler; s3) performing second mixing on the second culture medium filler and the bacterial liquid of aerobic bacteria to obtain final culture medium filler; wherein the first nutrient solution is a glucose solution; the second nutrient solution is a peptone solution. The method can solve the problem of low dredged sediment treatment efficiency in the prior art, and is suitable for the field of dredged sediment treatment.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of dredged sediment treatment, in particular to a preparation method of culture medium filler and application thereof. BACKGROUND

[0002] The treatment of river and lake dredged sediment has increasingly become an important issue in the field of environmental engineering. The composition of dredged sediment is significantly different from that of activated sludge, and the number of intrinsic beneficial microorganisms in dredged sediment is extremely limited. Direct application of traditional activated sludge aerobic fermentation technology for treatment faces many challenges. The content of nutrients in dredged sediment is low, and its composition is complex, which cannot meet the nutritional needs of microorganisms in the initial growth and reproduction. This characteristic promotes the ecological dominance of anaerobic microorganisms in the sediment, which decomposes organic matter into small molecules and produces methane, hydrogen sulfide and other flammable and toxic gases. The flammable nature of methane not only poses potential fire and explosion risks for engineering implementation, but the release of toxic gases such as hydrogen sulfide also poses a threat to the surrounding environment and the health of operating personnel. In addition, the high impurity content of the mixed gas produced during anaerobic fermentation makes it difficult to be effectively utilized, reducing the economic and feasibility of resource recovery.

[0003] In order to overcome the above difficulties and achieve effective aerobic fermentation of dredged sediment, the introduction of specific aerobic fermentation strains becomes a key strategy. The introduction of these strains aims to promote the rapid decomposition of organic matter in the sediment and inhibit the activity of anaerobic microorganisms, thereby avoiding the generation of toxic and harmful gases. However, the existing methods of introducing strains, whether by spraying liquid bacterial agents or mixing powdered bacterial agents, do not fully consider the uneven distribution of organic matter in dredged sediment. This oversight results in the inability of aerobic microorganisms to quickly locate and utilize organic nutrients in the sediment, limiting their growth and activity, which affects the efficiency of the early fermentation process and results in low efficiency of dredged sediment treatment. SUMMARY

[0004] The main purpose of the present application is to provide a preparation method of culture medium filler and application thereof, in order to solve the problem of low efficiency of dredged sediment treatment in the prior art.

[0005] In order to achieve the above purpose, according to the first aspect of the present application, a preparation method of culture medium filler is provided, which comprises: S1) sequentially soaking an initial culture medium filler in a first nutrient solution and a second nutrient solution to obtain a first culture medium filler; S2) first mixing the first culture medium filler with a liquid of shiitake mushroom to obtain a second culture medium filler; S3) second mixing the second culture medium filler with a liquid of aerobic bacteria to obtain a final culture medium filler; wherein the first nutrient solution is a glucose solution; and the second nutrient solution is a protein peptone solution.

[0006] Further, S1) comprises: S1-1) performing first soaking of the initial culture medium filler in a glucose solution to obtain a first soaked culture medium filler; S1-2) performing second soaking of the first soaked culture medium filler in a proteose peptone solution after first air-drying of the first soaked culture medium filler to obtain a second soaked culture medium filler; and S1-3) performing second air-drying of the second soaked culture medium filler to obtain the first culture medium filler; preferably, the glucose solution has a mass concentration of 5-20%; and preferably, the proteose peptone solution has a mass concentration of 0.1-5%.

[0007] Further, the first soaking time is 23.5-24.5 h; preferably, the first air-drying temperature is 19.5-20.5℃; preferably, the first air-drying time is 23.5-24.5 h; preferably, the second air-drying temperature is 19.5-20.5℃; and preferably, the second air-drying time is 1.75-2.25 h.

[0008] Further, the living bacteria content in the bacteria solution of the Lentinula edodes is greater than or equal to 10 billion / g; preferably, the first mixing time is 23.5-24.5 h; and preferably, the first mixing temperature is 25-30.5℃.

[0009] Further, the aerobic bacteria comprise one or more of Bacillus subtilis, thermophilic fungi or thermophilic actinomycetes; preferably, the living bacteria content in the bacteria solution of the Bacillus subtilis is greater than or equal to 200 billion / g; preferably, the living bacteria content in the bacteria solution of the thermophilic fungi is greater than or equal to 200 billion / g; and preferably, the living bacteria content in the bacteria solution of the thermophilic actinomycetes is greater than or equal to 200 billion / g.

[0010] Further, the preparation method of the initial culture medium filler comprises: combining the gap on the first mortise-tenon structure with the gap on the second mortise-tenon structure, and then combining the gaps of the two fixed structures with the two sides of the first mortise-tenon structure to obtain the initial culture medium filler.

[0011] Further, S3) comprises: performing second mixing of the second culture medium filler with the bacteria solution of the aerobic bacteria to obtain a second mixed culture medium filler; and performing third air-drying of the second mixed culture medium filler to obtain the final culture medium filler.

[0012] Further, the second mixing time is 0.5-1 min.

[0013] Further, the third air-drying time is 1±0.2 h; and preferably, the third air-drying temperature is 14.5-15.5℃.

[0014] In order to achieve the above-mentioned purpose, according to the second aspect of the present application, the preparation method of the culture medium filler is provided for application in the treatment of dredged sludge.

[0015] The technical scheme of the present application is used in the preparation of the culture medium filler for the treatment of the dredged sludge for carrying aerobic bacteria, the initial culture medium filler is soaked in the glucose solution and the protein solution in sequence, so that the initial culture medium filler has the growth conditions of the aerobic bacteria, and then the initial culture medium filler is soaked in the lentinus edodes, the growth conditions of the aerobic bacteria on the filler in the later period are improved, the growth and activity of the aerobic bacteria are promoted, and the treatment efficiency of the dredged sludge is further improved. BRIEF DESCRIPTION OF DRAWINGS

[0016] The drawings accompanying the specification of the present application form a part thereof, serve to provide further understanding of the present application, and together with the description of the present application, serve to explain the present application. In the drawings:

[0017] Figure 1 The structure diagram of the first mortise and tenon structure 1, the second mortise and tenon structure 2 and the fixing structure 3 of the initial culture medium filler assembly in the specification of the present application is shown.

[0018] Figure 2 The structure diagram of the initial culture medium filler in the specification of the present application is shown.

[0019] Figure 3 The flow diagram of the embodiment in the specification of the present application is shown.

[0020] Figure 4 The statistical diagram of the growth speed of the miscellaneous bacteria in group F in embodiment 4 in the specification of the present application is shown.

[0021] Figure 5 The statistical diagram of the growth speed of the miscellaneous bacteria in group G in embodiment 4 in the specification of the present application is shown.

[0022] The reference numerals in the drawings are specifically represented as: 1: first mortise and tenon structure; 2: second mortise and tenon structure; 3: fixing structure. DETAILED DESCRIPTION

[0023] It should be noted that the embodiments in the present application and the features in the embodiments can be combined with each other without conflict. The present application will be described in detail below in combination with the embodiments.

[0024] As mentioned in the background, due to the uneven distribution of organic matter in the dredged sludge, the aerobic microorganisms are difficult to absorb the organic nutrients in the sludge, which hinders the growth and development of the aerobic microorganisms, limits the reproduction and activity of the aerobic microorganisms in the dredged sludge, and reduces the treatment efficiency of the dredged sludge. Based on this, the present application attempts to provide a new preparation method of the culture medium filler, and thus a series of protection schemes of the present application are proposed.

[0025] In the first typical embodiment of the present application, a preparation method of culture medium filler is provided, which comprises: S1) sequentially immersing an initial culture medium filler in a first nutrient solution and a second nutrient solution to obtain a first culture medium filler; S2) performing a first mixing of the first culture medium filler with a Lentinula edodes bacterial solution to obtain a second culture medium filler; S3) performing a second mixing of the second culture medium filler with an aerobic bacteria bacterial solution to obtain a final culture medium filler; wherein the first nutrient solution is a glucose solution; and the second nutrient solution is a protein peptone solution.

[0026] The preparation method of the culture medium filler in the present application aims to solve the problems of uneven nutrition and poor distribution of aerobic bacteria in the initial stage of dredged sediment fermentation. The step-by-step nutrition strengthening and strain introduction process helps to promote the rapid proliferation and activity of aerobic microorganisms in the sediment in the initial stage of dredged sediment fermentation, providing strong support for subsequent aerobic fermentation.

[0027] In step S1), the initial culture medium filler undergoes pre-soaking treatment, and through the ordered application of the first nutrient solution (glucose solution) and the second nutrient solution (protein peptone solution), a first culture medium filler rich in carbon and nitrogen sources is formed. Glucose as a carbon source provider can be rapidly metabolized by microorganisms, promoting their early growth and not promoting excessive growth of miscellaneous bacteria; while protein peptone as a nitrogen source is crucial for building microbial protein matrix, helping to diversify and stably grow microbial populations. This two-stage nutrient solution soaking strategy not only enhances the nutrition supply of the culture medium, but also ensures that the solution can uniformly soak the filler, creating favorable conditions for the early attachment and proliferation of microorganisms.

[0028] In step S2), through the first mixing with the Lentinula edodes bacterial solution, the first culture medium filler is endowed with additional biological activity. As one of the dominant strains, Lentinula edodes is pre-activated in the glucose and protein peptone nutrient solution, which can more effectively work with aerobic bacteria to improve the overall metabolic efficiency. This process not only improves the survival rate of aerobic bacteria, but also promotes the interaction between bacterial populations, forming a healthier microbial ecology.

[0029] The role of Lentinula edodes in the culture medium filler of the present application is as follows: in the early stage of aerobic fermentation, Lentinula edodes preferentially releases heat through respiration, increasing the overall temperature of the fermentation pile, activating aerobic bacteria; on the other hand, Lentinula edodes can partially decompose organic pollutants in dredged sediment, promoting the maturation of the sediment; finally, in the middle and late stages of fermentation, Lentinula edodes enters a dormant state to provide nutrients for aerobic microorganisms. The Lentinula edodes in the culture medium filler of the present application can be any conventional commercially available product, all of which can achieve the effects of the present application.

[0030] In step S3), the second culture medium filler is mixed with the aerobic bacteria liquid to complete the final microbial inoculation process. The aerobic bacteria are aerobic bacteria known to those skilled in the art, including but not limited to Bacillus subtilis, thermophilic fungi and thermophilic actinomycetes. When the aerobic bacteria are planted in the culture medium filler treated in the present application, the contact probability of the microorganisms and the organic matter in the sediment can be improved, and the uneven problem commonly seen in traditional spraying or mixing methods can be avoided. Through the preparation method of the culture medium filler in the present application, the aerobic microorganisms carried by the culture medium filler enter the dredged sediment, not only solving the problem of energy and nutrient supply of aerobic bacteria in the early stage of sediment fermentation, but also further improving the fermentation efficiency and the quality of the product, overcoming the obstacles of aerobic fermentation in sediment treatment, realizing the efficient use of microorganisms and the improvement of fermentation performance, reducing the application amount of aerobic fermentation bacteria, reducing the cost of dredged sediment treatment, and opening up a new path for the environment-friendly treatment and resource utilization of dredged sediment.

[0031] In a preferred embodiment, S1) comprises: S1-1) first soaking the initial culture medium filler in a glucose solution to obtain a first soaked culture medium filler; S1-2) after first air-drying the first soaked culture medium filler, second soaking the first air-dried culture medium filler in a protein peptone solution to obtain a second soaked culture medium filler; S1-3) second air-drying the second soaked culture medium filler to obtain the first culture medium filler; preferably, the mass concentration of the glucose solution is 5-20% (including but not limited to 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% or 20%); preferably, the mass concentration of the protein peptone solution is 0.1-5% (including but not limited to 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4% or 5%).

[0032] The present application improves the activity and fermentation efficiency of microorganisms by regulating the nutrition process of the filler. The first soaking of the initial culture medium filler in the glucose solution provides the microorganisms with an easily absorbed carbon source, promoting their early metabolic activity. The volume concentration of the glucose solution is preferably set at 5-20%, which can not only meet the nutritional needs of the microorganisms, but also avoid the inactivation of the microorganisms due to too high concentration. By first air-drying the culture medium filler after the first soaking, and then second soaking in a protein peptone solution, the supply of carbon and nitrogen sources is combined. The volume concentration of the protein peptone solution is preferably 0.1-5%, which can not only meet the nitrogen requirements of the microorganisms for protein synthesis, but also balance the stability of the solution and the long-term growth needs of the microorganisms. The present application also controls the parameters of the two air-drying processes to avoid damage to the microorganisms due to excessive dehydration, and prevents the risk of corruption caused by residual nutrient solution.

[0033] In a preferred embodiment, the first soaking time is 23.5-24.5h (including but not limited to 23.5, 24 or 24.5h); preferably, the first air-drying temperature is 19.5-20.5℃ (including but not limited to 19.5℃, 20℃ or 20.5℃); preferably, the first air-drying time is 23.5-24.5h (including but not limited to 23.5, 24 or 24.5h); preferably, the second air-drying temperature is 19.5-20.5℃ (including but not limited to 19.5℃, 20℃ or 20.5℃); preferably, the second air-drying time is 1.75-2.25h (including but not limited to 1.75, 1.76, 1.78, 1.79, 1.8, 1.81, 1.82, 1.83, 1.84, 1.85, 1.86, 1.87, 1.88, 1.89, 1.9, 1.91, 1.92, 1.93, 1.94, 1.95, 1.96, 1.97, 1.98, 1.99, 2.00, 2.10, 2.11, 2.12, 2.13, 2.14, 2.15, 2.20, 2.21, 2.22, 2.23, 2.24 or 2.25h).

[0034] The present application controls the parameters in each of the above fermentation conditions precisely, so that they coordinate with each other to provide a good growth environment for the microorganism, which helps to maintain the activity of the microorganism and improve the overall efficiency of the fermentation. The present application controls the first soaking time to be 23.5-24.5 hours, so that the microorganism has enough time to absorb the nutritional ingredients in the glucose solution, while avoiding excessive consumption of solutes or the microorganism entering the decline phase too early due to too long soaking. The present application controls the first air-drying temperature to be 19.5-20.5℃ and the time to be 23.5-24.5 hours, which can effectively remove the excess moisture on the surface of the culture medium filler through the first air-drying, avoid the microorganism being in an unfavorable environment with too high humidity, while retaining the necessary humidity to maintain the active state of the microorganism. Furthermore, the present application shortens the second air-drying time to 1.75-2.25 hours, which aims to quickly remove the moisture in the peptone solution, prevent the microorganism from being exposed to a dry environment for too long and causing a decrease in activity, and at the same time ensure that the nutrient solution is fully retained inside the filler to create conditions for the next step of microorganism inoculation. In a preferred embodiment, the viable bacteria content in the Lentinula edodes bacterial solution is ≥1 billion / g; preferably, the first mixing time is 23.5-24.5h; preferably, the first mixing temperature is 25-30.5℃.

[0035] In a preferred embodiment, the aerobic bacteria include, but are not limited to, one or more of Bacillus subtilis, thermophilic fungi or thermophilic actinomycetes. The aerobic bacteria in the culture medium filler of the present application can also be any aerobic bacteria known to those skilled in the art for use in sediment dredging projects. The above-mentioned Bacillus subtilis, thermophilic fungi and thermophilic actinomycetes can be selected from any conventional commercially available product, all of which can achieve the effects of the present application.

[0036] In a preferred embodiment, the viable bacterial content of the Bacillus subtilis bacterial solution is ≥200 billion / g; the viable bacterial content of the thermophilic fungi bacterial solution is ≥200 billion / g; and the viable bacterial content of the thermophilic actinomycetes bacterial solution is ≥200 billion / g.

[0037] By controlling the viable bacterial content of the Lentinula edodes bacterial solution in the above range, the present application can quickly establish a bacterial advantage when mixed with the culture medium filler, promoting efficient decomposition of organic matter. The first mixing time is preferably set to 23.5-24.5 hours. Compared with the traditional short mixing method, this can give the bacterial solution and the filler more sufficient contact opportunities, promote the uniform distribution and preliminary adaptation of microorganisms, and thus enhance the starting efficiency of the entire fermentation system. At the same time, the present application controls the environmental temperature of the first mixing to be 25-30.5°C, which can promote the metabolic activity of Lentinula edodes and accelerate the utilization speed of nutrients by microorganisms, laying a foundation for the subsequent fermentation process.

[0038] Further, the aerobic bacteria of the present application include, but are not limited to, Bacillus subtilis, thermophilic fungi and thermophilic actinomycetes, each of which has a viable bacterial content of not less than 200 billion / g, which can strengthen the decomposition of microorganisms, help the culture medium filler of the present application overcome the problem of uneven distribution of organic matter in the sediment, improve the activity and decomposition efficiency of microorganisms in the sediment fermentation process, achieve dynamic balance of the microbial community, reduce the generation of miscellaneous bacteria, and make up for the low utilization efficiency of microorganisms in the prior art in the field of dredged sediment treatment.

[0039] In a preferred embodiment, the preparation method of the initial culture medium filler comprises: combining the gap on the first mortise-tenon structure with the gap on the second mortise-tenon structure, and then combining the gaps of the two fixed structures with the two sides of the first mortise-tenon structure to obtain the initial culture medium filler.

[0040] The structural diagram of the first mortise-tenon structure 1, the second mortise-tenon structure 2 and the fixed structure 3 in the present application is shown in Figure 1 The structure of the initial culture medium filler formed after assembling these structures is shown in Figure 2The preparation method of the present application can not only be used to prepare the final medium filler of the present application by using the initial medium filler, but also can be used to prepare the medium filler and improve the activity of the bacterial strain, thereby improving the fermentation efficiency and the treatment efficiency of the dredged sediment, by using the medium filler for the treatment of the dredged sediment known to those skilled in the art.

[0041] In a preferred embodiment, S3) comprises obtaining a second mixed medium filler by mixing the second medium filler with the bacterial solution of the aerobic bacteria; and obtaining the final medium filler by thirdly drying the second mixed medium filler.

[0042] In a preferred embodiment, the second mixing time is 0.5-1 min (including but not limited to 0.5, 0.6, 0.7, 0.8, 0.9 or 1 min). Preferably, the second mixing temperature is room temperature, i.e. 15-20°C (including but not limited to 15°C, 16°C, 17°C, 18°C, 19°C or 20°C).

[0043] In a preferred embodiment, the third drying time is 1±0.2 h (including but not limited to 0.8, 0.9, 1.0, 1.1 or 1.2 h); and preferably, the third drying temperature is 14.5-15.5°C (including but not limited to 14.5°C, 14.6°C, 14.7°C, 14.8°C, 14.9°C, 15.0°C, 15.1°C, 15.2°C, 15.3°C, 15.4°C or 15.5°C).

[0044] The present application can efficiently combine the bacterial strain and the medium filler and stably maintain the activity of the microorganism by precisely controlling the second mixing and the third drying. The second mixing time is controlled to be 0.5-1 min, which aims to prevent the local concentration from being too high due to the long residence of the bacterial solution on the surface of the filler, thereby affecting the uniform distribution of the aerobic bacteria and the consistency of the activity. In cooperation therewith, the second mixed medium filler is then subjected to the third drying, the time of which is limited to 1 h and the temperature of which is controlled to be 14.5-15.5°C. This rapid and low-temperature drying strategy can not only rapidly remove the excess water of the bacterial solution and prevent the excessive dehydration of the microorganism, but also can avoid the decrease of the activity of the bacterial strain caused by high temperature and improve the fermentation efficiency of the microorganism in the subsequent fermentation process.

[0045] In a second typical embodiment of the present application, the application provides an application of the above-mentioned preparation method of the medium filler in the treatment of the dredged sediment. Unless otherwise specified, the reagents in the embodiments of the present application are all conventional commercially available products.

[0046] The beneficial effects of the present application will be further explained in detail below with reference to specific embodiments.

[0047] Example 1

[0048] The flow chart of the embodiment of the present application is shown as Figure 3

[0049] 1. Prepare the initial culture medium filler, and the specific preparation method comprises the following steps:

[0050] S1. Pine is made into a pine board with a thickness of 1 cm, and then a first mortise and tenon structure, a second mortise and tenon structure and two fixing structures are cut out by laser cutting.

[0051] S2. The first mortise and tenon structure and the second mortise and tenon structure are assembled through a gap, and the relatively sharp parts are combined to form a sharp head during assembly.

[0052] S3. The two fixing structures are fixed from opposite sides, and the gap of the fixing structure is combined with the two sides of the first mortise and tenon structure to reinforce the whole structure, and an initial culture medium filler with a total weight of 30 kg is obtained.

[0053] 2. The initial culture medium filler is poured into a 10% concentration glucose solution and soaked for 24 h.

[0054] After soaking is completed, it is placed in a forced air drying oven and the temperature of the drying oven is kept at 20℃, and after air drying for 24 h, it is taken out. The dried filler is soaked in 1% peptone nutrient solution for 4 h, and after soaking for 2 h, a first culture medium filler is obtained. The first culture medium filler is continuously placed in a forced air drying oven and the temperature of the drying oven is kept at 20℃, and after air drying for 2 h, it is taken out.

[0055] 3. The dried filler is coated with a mushroom fungus liquid (Wuhan Ruideng Biomaterial Technology Co., Ltd. BNCC336091) on the surface with a brush, and the filler coated with the fungus liquid is placed in a sterile drying oven for 24 h, and the temperature is kept at 27±2℃, to obtain a second culture medium filler.

[0056] 4. The second culture medium filler is placed in the fungus liquid of Bacillus subtilis, thermophilic fungi and thermophilic actinomycetes (fungus liquid of aerobic bacteria) after standing for 24 h.

[0057] The fungus liquid is obtained by mixing fungus powder and water, 10 g of fungus powder is added to 1 L of water, and the number of live bacteria of Bacillus subtilis (Hebei Hongtao Biological Engineering Co., Ltd.), thermophilic fungi (Wuhan Ruideng Biomaterial Technology Co., Ltd. CICC10267) and thermophilic actinomycetes (Wuhan Ruideng Biomaterial Technology Co., Ltd. CICC10347) in the fungus powder is not less than 200 billion / g, the filler is taken out after soaking for 1 min, and is subjected to rapid air drying in a forced air drying oven, and the air drying time is 1 h.

[0058] ​The dried filler is the final culture medium filler, and the auxiliary material (wood chips) is mixed with the sediment during aerobic fermentation of the sediment, and a small amount of water is added during mixing to make the overall moisture content reach 80%.

[0059] Example 2

[0060] In the experiment with Lentinula edodes as the dominant fermentation strain, a blank control group is designed to verify the necessity of Lentinula edodes. The specific experimental design is as follows:

[0061] The initial culture medium filler is first poured into a 10% glucose solution and soaked for 24 hours. After soaking, it is placed in a forced air drying oven and the temperature of the drying oven is maintained at 20℃. After drying for 24 hours, it is taken out. The dried filler is then soaked in a 1% proteose peptone nutrient solution for 4 hours. After 2 hours of soaking, it is placed in a forced air drying oven again and the temperature of the drying oven is maintained at 20℃. After drying for 2 hours, it is taken out.

[0062] The filler is then divided into two groups, A and B. Group A is coated with Lentinula edodes liquid (prepared according to the method of Example 1) using a brush, and group B is coated with distilled water. The coated fillers are placed separately in sterile drying ovens for 24 hours, with the temperature maintained at 27℃. After 24 hours of standing, each of groups A and B is divided into three parts, and each part is coated with the same concentration of Bacillus subtilis, thermophilic fungi, and thermophilic actinomycete liquid (prepared according to the method of Example 1) on the surface. The coated fillers are placed in a ventilated environment at 50℃ for 48 hours, and then the number of Bacillus subtilis, thermophilic fungi, and thermophilic actinomycetes is counted. The test results are shown in Table 1.

[0063] Table 1 Effect of Lentinula edodes on aerobic microorganisms

[0064] Group number Bacillus subtilis / (h CFU) Thermophilic fungi / (h CFU) Thermophilic actinomycetes / (h CFU) A 548 327 416 B 125 106 209

[0065] According to the above table, the addition of Lentinula edodes can effectively increase the number of Bacillus subtilis, and also has a significant promoting effect on the activity of thermophilic fungi and thermophilic actinomycetes.

[0066] Example 3

[0067] The use of glucose and proteose peptone as culture media is tested to verify the necessity of the two substances. The specific experimental steps are as follows: three groups of culture dishes are taken and designated as groups C, D, and E.

[0068] Group C: The initial culture medium filler prepared in Example 1 is poured into a 10% glucose solution 10g;

[0069] Group D: The initial culture medium filler prepared in Example 1 is poured into a 1% proteose peptone nutrient solution 10g;

[0070] Group E: The initial culture medium prepared in Example 1 was poured into 5 g of 10% glucose solution and 5 g of 1% peptone nutrient solution;

[0071] Then, the three groups of culture dishes were respectively inoculated with the same mass concentration (5‰) of Lentinula edodes suspension, and cultured at 37±0.1°C. After 24 hours of culture, the number of Lentinula edodes was counted. The test results are shown in Table 2.

[0072] Table 2 Influence of medium composition on Lentinula edodes

[0073] Group number Lentinula edodes / (h CFU) C 179 D 104 E 387

[0074] According to the above table, the number of Lentinula edodes cultured in group D is the least, the number of Lentinula edodes cultured in group C is slightly more, and the number of Lentinula edodes cultured in group E is the most. This shows that the use of two kinds of nutrient solution is the best for Lentinula edodes. However, in practical application, the use of a single nutrient substance can reduce costs, but it will obviously affect the fermentation effect.

[0075] Example 4

[0076] In order to verify the influence of the soaking order of peptone and glucose on the inoculation of strains in the later stage, control experiments were designed as group F and group G.

[0077] Group F: Consistent with Example 1, first soak the glucose solution, then soak the peptone solution;

[0078] Group G: The difference from Example 1 is that the peptone solution is soaked first, and then the glucose solution is soaked. Except for the order, the remaining variables are consistent with Example 1.

[0079] Then, the soaked medium was smeared with an equal amount of Lentinula edodes, and then placed in a ventilated environment with 95% humidity and 25°C. The growth of Lentinula edodes and miscellaneous bacteria was observed. The growth rates of miscellaneous bacteria under different soaking orders are shown in Figure 4 (F group) and Figure 5 (G group), Figure 4 and Figure 5 The first day in each of them means the results after 1 day of fermentation.

[0080] According to the cases of Figure 4 and Figure 5 , soaking the peptone solution first is more conducive to the growth of miscellaneous bacteria. After 6 days of culture, Lentinula edodes has basically died. Therefore, soaking glucose first is more beneficial to the planting of strains in the later stage.

[0081] Example 5

[0082] A small reservoir in the north is taken as an example. Large pastures and farms are built on both sides of the upstream of the reservoir, resulting in a large amount of organic matter in the reservoir sediment. The pH, organic matter, soil infiltration rate, hydrolytic nitrogen, available phosphorus, available potassium and germination index of the sediment are shown in Table 3. In order to verify the effect of the filler, an engineering test of 500 tons of dredged sediment is carried out.

[0083] The water content of the existing 500 tons of dredged sediment is 72.19%. According to the ratio of sediment: culture medium filler = 7:3, 214 tons of culture medium filler are needed. According to the preparation method of the initial culture medium filler in Example 1, the initial culture medium filler is prepared by poplar according to the local conditions.

[0084] According to the quality and volume of the filler, 100 tons of 10% glucose solution, 50 tons of 1% protein peptone solution, 12 tons of 1×10 9 Bacillus subtilis, thermophilic fungi and thermophilic actinomycetes are prepared.

[0085] According to the specific implementation steps of Example 1, the 214 tons of culture medium filler are divided into 5 batches for soaking. The solution concentration or microbial activity should be detected before each batch of soaking. If it does not meet the requirements, appropriate replacement or addition of corresponding supplements should be made.

[0086] After mixing the prepared culture medium filler with the dredged sediment, it is sent to the fermentation warehouse for fermentation. After fermentation, the product index and the index before fermentation are compared as shown in Table 3.

[0087] The detection standards of each index are detected according to the records in “Organic Fertilizer” (NY / T525-2021).

[0088] Table 3 Comparison of key indicators before and after fermentation

[0089] Serial number Detection item Unit Index before fermentation Index of fermentation product 1 pH / 7.57 7.89 2 Organic matter g / kg 35.9 51.3 3 Soil infiltration rate mm / h 2.4 5.21 4 Hydrolytic nitrogen mg / kg 120 198 5 Available phosphorus mg / kg 2.59 6.94 6 Available potassium mg / kg 42.8 247.2 7 Germination index ％ 45 92

[0090] As shown in the above table, the product fermented by the culture medium filler of the application can effectively improve the infiltration rate, organic matter content, hydrolytic nitrogen, available phosphorus, available potassium and other indicators of the dredged sediment, and the seed germination rate is also obviously improved, which is conducive to the use of dredged sediment as plant cultivation substrate.

[0091] From the above description, it can be seen that the above-mentioned embodiments of the present application achieve the following technical effects: the present application improves the inoculation efficiency and distribution uniformity of aerobic microorganisms by accurately regulating the viable bacterial content and mixing conditions of the bacterial strain, enhances the decomposition ability of microorganisms to organic matter in dredged sediment, shortens the fermentation start-up time, and optimizes the fermentation cycle. The culture medium filler preparation method of the present application solves the problems of low bacterial activity and unstable fermentation effect in traditional aerobic fermentation, improves the economic efficiency and environmental friendliness of dredged sediment treatment, opens up a new way for the resource utilization of sediment, and shows the potential and practical application value in the field of microbial fermentation technology.

[0092] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. For those skilled in the art, the present application can have various modifications and changes. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. A method for the preparation of a culture medium filler, characterized in that, The preparation method comprises: S1) sequentially immersing an initial culture medium filler in a first nutrient solution and a second nutrient solution to obtain a first culture medium filler; S2) performing first mixing of the first culture medium filler with a fungus liquid of a lentinus edodes to obtain a second culture medium filler; S3) performing second mixing of the second culture medium filler with a fungus liquid of an aerobic bacteria to obtain the final culture medium filler; The first nutrient solution is a glucose solution; The second nutrient solution is a proteose peptone solution.

2. The production method according to claim 1, characterized by, The S1) comprises: S1-1) performing first immersion of the initial culture medium filler in the glucose solution to obtain a first immersion culture medium filler; S1-2) performing first air drying of the first immersion culture medium filler and then performing second immersion of the first immersion culture medium filler in the proteose peptone solution to obtain the second immersion culture medium filler; S1-3) performing second air drying of the second immersion culture medium filler to obtain the first culture medium filler; Preferably, the mass concentration of the glucose solution is 5-20%; Preferably, the mass concentration of the proteose peptone solution is 0.1-5%.

3. The preparation method according to claim 2, characterized in that, The first immersion time is 23.5-24.5 h; Preferably, the first air drying temperature is 19.5-20.5℃; Preferably, the first air drying time is 23.5-24.5 h; Preferably, the second air drying temperature is 19.5-20.5℃ Preferably, the second air drying time is 1.75-2.25 h.

4. The preparation method according to claim 1, characterized in that, The live bacteria content in the fungus liquid of the lentinus edodes is ≥10 billion / g; Preferably, the first mixing time is 23.5-24.5 h; Preferably, the first mixing temperature is 25-30.5℃.

5. The preparation method according to claim 1, characterized in that, The aerobic bacteria comprise one or more of bacillus subtilis, thermophilic fungi or thermophilic actinomycetes; Preferably, the live bacteria content in the fungus liquid of the bacillus subtilis is ≥200 billion / g; Preferably, the live bacteria content in the fungus liquid of the thermophilic fungi is ≥200 billion / g; Preferably, the live bacteria content in the fungus liquid of the thermophilic actinomycetes is ≥200 billion / g.

6. The method of claim 5, wherein the step of forming the first and second layers is performed by a method comprising: The preparation method of the initial culture medium filler comprises: combining a gap on a first mortise-tenon structure with a gap on a second mortise-tenon structure, and then combining the gaps of two fixing structures with the two sides of the first mortise-tenon structure to obtain the initial culture medium filler.

7. The preparation method according to claim 1, characterized in that, The S3) comprises performing second mixing of the second culture medium filler with the fungus liquid of the aerobic bacteria to obtain a second mixed culture medium filler; Performing third air drying of the second mixed culture medium filler to obtain the final culture medium filler.

8. The preparation method according to claim 7, characterized in that, The second mixing time is 0.5-1 min.

9. The preparation method according to claim 7, characterized in that, The third air drying time is 1±0.2 h; Preferably, the third air drying temperature is 14.5-15.5℃.

10. Application of the preparation method of the culture medium filler in any one of claims 1-9 in the treatment of dredged mud.

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