Preparation method of composite mildew removing agent and composite mildew removing agent

Abstract

The invention relates to the technical field of mold removal agents, in particular to a preparation method of a composite mold removal agent and the composite mold removal agent. The preparation method of the composite mildew removing agent comprises the following steps: S1, preparing modified montmorillonite; s2, brevibacillus laterosporus is cultured in a culture medium, brevibacillus laterosporus liquid is obtained after culture is completed, and then the obtained brevibacillus laterosporus liquid is prepared into dry powder; s3, mixing methyl thioglycolate, dimethyl trisulfide, 3-methylbutyl sulfate S-methyl ester and water, and then adding the dry powder to obtain a mixed solution; s4, mixing the mixed solution with the modified montmorillonite to obtain a composite mildew removing agent preparation; and S5, drying the composite mildew removing agent, and packaging. The composite mildew removing agent is applied to mildewed feed or corn starch, and an excellent mildew removing effect can be achieved 24 hours after mildew removing treatment.

Description

Technical Field

[0001] The present application relates to the technical field of mold removers, and in particular to a preparation method of a composite mold remover and the composite mold remover. Background Art

[0002] Mycotoxins are toxic metabolites produced by mold in contaminated grains. The effects of mycotoxins on grains can be categorized into four main aspects:

[0003] Impact on grain quality: Mycotoxin contamination can cause grain deterioration, reduce its edible value, and cause economic losses. Mycotoxin contamination can also cause grain to become moldy, discolored, or have an odor, seriously affecting its appearance and taste.

[0004] Impact on human health: After people ingest large doses of mycotoxins, they may experience acute pathological changes such as liver parenchymal cell necrosis, bile duct epithelial cell hyperplasia, liver fat infiltration and liver hemorrhage. The early symptoms are fever, vomiting, anorexia, jaundice, followed by ascites, lower limb edema and death soon after. Long-term intake of grains containing low doses of mycotoxins may cause chronic harm to human health, such as increasing the risk of cancer and affecting the function of the immune system.

[0005] Impact on livestock and poultry farming: Using grains contaminated with mycotoxins as feed will reduce livestock and poultry's feed intake and feed utilization rate, leading to slow growth and weight loss. Mycotoxins may also damage the reproductive system of livestock and poultry, leading to poor reproductive performance, such as estrus in sows, anestrus in sows, no estrus in sows, small litters in sows, miscarriage in sows, stillbirth in sows, low libido in boars, low sperm motility in boars, and decreased semen quality in boars. Mycotoxins can also remain in livestock and poultry products, posing a huge threat to human health and safety.

[0006] Impact on the agricultural economy: Mycotoxin contamination can cause grain losses and reduced quality, severely impacting agricultural economic development. Furthermore, mycotoxin-contaminated feed can lead to significant economic losses in livestock and poultry, leading to illness and death.

[0007] According to statistics, there are more than 300 known mycotoxins, among which aflatoxin, zearalenone, T-2 toxin, ochratoxin, vomitoxin, fumonisin, etc. are highly toxic, widely distributed, and have high toxin production.

[0008] Methods for removing mycotoxins include chemical degradation, physical degradation, and biological degradation. Among them, the chemical degradation method mainly uses oxidants or strong alkalis to degrade toxins. Physical degradation methods mainly include adsorbent water washing, adsorption, solvent extraction, heating detoxification, ultraviolet detoxification, and germ removal, among which the most commonly used is adsorbent adsorption. However, the above two methods have the defects of unstable and incomplete toxin degradation effects, and will lead to the loss of nutrients and trace elements, poor palatability, and other shortcomings. The biological degradation method mainly uses microorganisms or their degradation products to degrade toxins. It has the advantages of having little impact on the sensory properties, palatability, and nutrients of the raw materials. At the same time, it is also safe, environmentally friendly, and efficient.

[0009] Chinese patent application number 202310154956.6 discloses a biological demycotoxin for degrading mycotoxins in feed. The agent was applied to initially moldy silage. After one week, the toxins and their contents were measured. The results showed that the proportion of moldy feed was significantly reduced. The degradation efficiency of zearalenone reached 89%, and that of vomitoxin reached 90%. Aflatoxin B1 was no longer detected. The applicant believes that existing methods for degrading mycotoxins with microorganisms take a long time to degrade, and the degradation rate needs to be improved. Summary of the Invention

[0010] In order to improve the degradation rate of mycotoxins and shorten the degradation time, the present application provides a preparation method of a composite mold removal agent and the composite mold removal agent.

[0011] In a first aspect, the present application provides a method for preparing a composite mold removal agent, which adopts the following technical solution.

[0012] A method for preparing a composite mold removal agent comprises the following steps:

[0013] S1. Preparation of modified montmorillonite:

[0014] 1) Dissolve copper sulfate in water to prepare copper sulfate solution;

[0015] 2) Add montmorillonite to the copper sulfate solution, and then add a pH adjuster to adjust the pH value of the solution to 10;

[0016] 3) Add formaldehyde solution as a reducing agent and start the chemical deposition reaction at 70-75°C for 12-14 hours;

[0017] 4) After the reaction is completed, filtering, washing, and drying to remove unreacted copper ions and residual reducing agent to obtain modified montmorillonite;

[0018] S2. Brevibacillus laterosporus was cultured in a culture medium, and after completion of the culture, a Brevibacillus laterosporus bacterial solution was obtained, and then the resulting bacterial solution was prepared into a dry powder;

[0019] S3 methyl thioglycolate, dimethyl trisulfide, 3-methylbutyl sulfate S- methyl ester was mixed with water, and then the dry powder was added to obtain a mixture;

[0020] S4. The mixture was mixed with modified montmorillonite to obtain a composite mold removal agent preparation;

[0021] S5. Dry the composite mold removal agent and then package it.

[0022] By employing the above-mentioned technical solution, B. laterosporus can produce antimicrobial peptides and polyketides, which have inhibitory effects on a variety of molds. Antimicrobial peptides can bind to lipopolysaccharide / lipid II molecules on the mold cell membrane, altering the cell membrane's permeability and thereby inhibiting mold growth. Polyketides inhibit mold reproduction by affecting gene pathways in the mold cell membrane, such as ATP synthesis, peptidoglycan biosynthesis, membrane transport, and cellular metabolism. B. laterosporus secretes chitinase, an enzyme that degrades chitin in the fungal cell wall, disrupting the mold's structural integrity and thereby inhibiting its growth. During its growth, B. laterosporus consumes nutrients from the environment, competing with the mold for limited nutrient resources, thereby limiting its growth and reproduction.

[0023] Methyl thioglycolate can disrupt microbial cell membranes, causing leakage of cell contents, thereby inhibiting or killing bacteria. Thiol groups can react with sulfhydryl enzymes in microbial enzyme systems, disrupting their normal metabolism and thereby inhibiting their growth. Methyl thioglycolate can also disrupt fungal cell membranes, affecting cell wall synthesis and leading to cell death. Furthermore, it can interfere with fungal metabolic processes, inhibiting spore formation and germination, thereby achieving a mildew-proofing effect.

[0024] Dimethyl trisulfide directly harms fungi by disrupting their cell structure, affecting their normal growth and reproduction, thereby achieving a mildew-proofing effect. It can penetrate into fungal cells, destroying their proteins and plasma membranes, leading to water loss and rendering them incapable of survival. It also prevents the extravasation of various ions, enzymes, coenzymes, and intermediates within the cells, creating a virtuous cycle and enhancing mildew-proofing effectiveness. It can react with chromosomal material in fungal cells, inhibiting chromosome division or causing mutations, thereby affecting cell division, growth, and morphology, thereby achieving its mildew-proofing effect.

[0025] 3-Methylbutane sulfate S-methyl ester can disrupt microbial cell membranes, causing leakage of cell contents, thereby inhibiting or killing bacteria. Its ester groups and sulfur atoms allow it to react with enzymes within microorganisms, disrupting their normal metabolism and thereby inhibiting their growth. It can also damage fungal cell membranes, affecting cell wall synthesis and leading to cell death. Furthermore, it can interfere with fungal metabolic processes, inhibiting spore formation and germination, thereby achieving a mildew-proofing effect.

[0026] Copper ions are grafted onto montmorillonite through a chemical deposition method. This not only improves the montmorillonite's adsorption of mold, but also promotes the removal efficiency of Brevibacillus sporogenes, methyl thioglycolate, dimethyl trisulfide, and 3-methylbutyl sulfate S-methyl ester. The modified montmorillonite has a strong adsorption capacity, capable of absorbing mold on feed. Furthermore, the modified montmorillonite can adsorb Brevibacillus laterosporus, methyl thioglycolate, dimethyl trisulfide, and 3-methylbutyl sulfate S-methyl ester, resulting in a more precise inhibition of Brevibacillus laterosporus, methyl thioglycolate, dimethyl trisulfide, and 3-methylbutyl sulfate S-methyl ester, rapidly killing mold. Brevibacillus laterosporus, methyl thioglycolate, dimethyl trisulfide, and 3-methylbutyrate S-methyl sulfate work synergistically. Methyl thioglycolate, dimethyl trisulfide, and 3-methylbutyrate S-methyl sulfate can quickly eliminate mycotoxins. At the same time, methyl thioglycolate, dimethyl trisulfide, and 3-methylbutyrate S-methyl sulfate have a certain inhibitory effect on the release of substances from Brevibacillus laterosporus, causing the mold-removing substances of Brevibacillus laterosporus to be slowly released, thereby extending the effective action time of the mold-removing agent and allowing the mold to be removed more thoroughly.

[0027] Furthermore, the concentration of the copper sulfate solution is 16-20 g / L; the weight ratio of copper sulfate to montmorillonite is 1:(5-12); and the weight ratio of formaldehyde to copper sulfate is (3-4):5.

[0028] Furthermore, the weight ratio of the copper sulfate to the montmorillonite is 1:(6-8).

[0029] Furthermore, hexadecyltrimethylammonium bromide is grafted onto the modified montmorillonite by the following grafting method:

[0030] a) dissolving cetyltrimethylammonium bromide in water to obtain a cetyltrimethylammonium bromide solution, and adding methyl p-toluenesulfonate to the cetyltrimethylammonium bromide solution to obtain a treatment solution;

[0031] b) adding the modified montmorillonite to the treatment solution, mixing, and reacting under condensation reflux at 70-85°C for 2-4 hours;

[0032] c) After the reaction is completed, the slurry is ultrasonically dispersed, the supernatant is removed by centrifugation, the precipitate is collected, and the precipitate is washed; the washed precipitate is dried at 80-90° C. to obtain hexadecyltrimethylammonium bromide-modified montmorillonite.

[0033] By adopting the above technical solution, hexadecyltrimethylammonium bromide is grafted to increase the interlayer distance of montmorillonite, which on the one hand further improves the adsorption effect of montmorillonite, and on the other hand allows water molecules to better enter the interlayer of montmorillonite, thereby improving the water solubility of montmorillonite.

[0034] Furthermore, the weight ratio of hexadecyltrimethyl bromide to methyl p-toluenesulfonate is 1:(0.1-0.3); the weight ratio of hexadecyltrimethyl bromide to modified montmorillonite is 1:(35-42).

[0035] Further, methyl thioglycolate, dimethyl trisulfide, 3-methylbutyl sulfate S-methyl ester, and 3-mercaptopropyltrimethoxysilane in S3 are mixed with water, and then the dry powder is added to obtain a mixed solution.

[0036] By adopting the above technical solution, 3-mercaptopropyltrimethoxysilane containing a thiol group is physically mixed with montmorillonite, and the colonization effect of montmorillonite on probiotics is enhanced by physical adsorption.

[0037] In a second aspect, the present application provides a composite mold removal agent, which adopts the following technical solution.

[0038] A composite mold removal agent comprises the following raw materials in parts by weight: 5-15 parts of Brevibacillus laterosporus, 15-25 parts of methyl thioglycolate, 10-20 parts of dimethyl trisulfide, 15-35 parts of 3-methylbutyl sulfate S-methyl ester, 15-30 parts of modified montmorillonite, 5-9 parts of 3-mercaptopropyltrimethoxysilane, and 20-40 parts of water.

[0039] Furthermore, the weight ratio of the sum of the weights of methyl thioglycolate, dimethyl trisulfide, and 3-methylbutyl sulfate S-methyl ester to the weight of Brevibacillus laterosporus is (8-12):1.

[0040] Furthermore, when applied to moldy grains or feed, the weight ratio of the moldy grain raw materials or feed to the composite de-mold agent is (400-500):1.

[0041] By adopting the above technical solution, the composite mold removal agent is applied to moldy grains or feed, such as corn flour, and a smaller amount of the composite mold removal agent can achieve a higher mold removal efficiency.

[0042] In summary, this application has the following beneficial effects:

[0043] In this application, a composite mold removal agent is prepared by compounding modified montmorillonite with Brevibacillus laterosporus, methyl thioglycolate, dimethyl trisulfide, and 3-methylbutyl sulfate S-methyl ester, and applied to moldy corn flour. After 24 hours of mold removal treatment, the removal rate of zearalenone in the moldy corn flour can reach 96.4-99.8%, the removal rate of aflatoxin B1 in the moldy corn flour can reach 96.6-99.9%, and the removal rate of vomitoxin in the moldy corn flour can reach 96.1-99.5%. DETAILED DESCRIPTION

[0044] The present application is further described in detail below with reference to the embodiments.

[0045] Preparation examples of raw materials and intermediates

[0046] raw material

[0047] The raw materials in the examples of this application can be obtained commercially:

[0048] Brevibacillus laterosporus is a commercially available product;

[0049] Copper sulfate, CuSO4·5H2O, analytical grade;

[0050] pH adjuster, 40% sodium hydroxide solution;

[0051] Hexadecyltrimethylammonium bromide, analytical grade;

[0052] Methyl p-toluenesulfonate, analytical grade;

[0053] Methyl thioglycolate, analytical grade;

[0054] Dimethyl trisulfide, analytical grade;

[0055] 3-Methylbutane sulfate S-methyl ester, analytical grade;

[0056] Modified montmorillonite, analytical grade;

[0057] 3-Mercaptopropyltrimethoxysilane, analytical grade.

[0058] Preparation Example

[0059] Preparation Example 1

[0060] A modified montmorillonite, the preparation method of which is as follows:

[0061] 1) Dissolve 200g of copper sulfate in water to prepare a copper sulfate solution with a concentration of 18g / L;

[0062] 2) adding 1400 g of montmorillonite to the copper sulfate solution obtained in step 1), and then adding a pH adjuster to adjust the pH value of the solution to 10; the weight ratio of copper sulfate to montmorillonite is 1:7;

[0063] 3) Add 20 L of 6 g / L formaldehyde solution as a reducing agent and start the chemical deposition reaction at 70°C for 12 hours;

[0064] 4) After the reaction is completed, filtering, washing, and drying to remove unreacted copper ions and reducing agent residues to obtain modified montmorillonite.

[0065] Preparation Example 2

[0066] Different from Preparation Example 1, in step 2) of Preparation Example 2, the amount of montmorillonite is 1200 g, and the weight ratio of copper sulfate to montmorillonite is 1:6.

[0067] Preparation Example 3

[0068] Different from Preparation Example 1, in step 2) of Preparation Example 3, the amount of montmorillonite is 1600 g, and the weight ratio of copper sulfate to montmorillonite is 1:8.

[0069] Preparation Example 4

[0070] Different from Preparation Example 1, in step 2) of Preparation Example 3, the amount of montmorillonite is 1000 g, and the weight ratio of copper sulfate to montmorillonite is 1:5.

[0071] Preparation Example 5

[0072] A hexadecyltrimethylammonium bromide-modified montmorillonite, the preparation method of which is as follows:

[0073] a) dissolving 100 g of cetyltrimethylammonium bromide in 1000 g of water to obtain a cetyltrimethylammonium bromide solution, and adding 10 g of methyl p-toluenesulfonate to the cetyltrimethylammonium bromide solution to obtain a treated solution;

[0074] b) adding 3500 g of modified montmorillonite to the treated solution, mixing, and reacting under condensation reflux at 80° C. for 3 h;

[0075] c) After the reaction is completed, the slurry is ultrasonically dispersed, the supernatant is removed by centrifugation, the precipitate is collected, and the precipitate is washed three times with distilled water and anhydrous ethanol in sequence; the washed precipitate is dried at 85° C. to obtain hexadecyltrimethylammonium bromide-modified montmorillonite.

[0076] Preparation Example 6

[0077] The difference from Preparation Example 5 is that in Preparation Example 6 b) 4200 g of modified montmorillonite was added to the treatment liquid.

[0078] Preparation Example 7

[0079] The difference from Preparation Example 5 is that in Preparation Example 7 b) 3200 g of modified montmorillonite was added to the treatment liquid.

[0080] Preparation Example 8

[0081] The difference from Preparation Example 5 is that in Preparation Example 8 a) 30 g of methyl p-toluenesulfonate was added to the hexadecyltrimethylammonium bromide solution.

[0082] Preparation Example 9

[0083] The difference from Preparation Example 5 is that in Preparation Example 9 a) 40 g of methyl p-toluenesulfonate was added to the hexadecyltrimethylammonium bromide solution.

[0084] Example

[0085] Examples 1-8

[0086] A composite mildew remover, the preparation method of which is as follows:

[0087] S1. Modified montmorillonite was prepared according to the method of Preparation Example 1;

[0088] S2 according to the ratio in Table 1 Brevibacillus laterosporus was cultured in a culture medium, and after completion of the culture, Brevibacillus laterosporus was obtained by bacterial solution, and then the obtained bacterial solution was prepared into a dry powder;

[0089] The culture medium for Brevibacillus laterosporus is: molasses 12-16g / L, beef powder 6-10g / L, magnesium sulfate 0.6-1g / L, KH2PO4 0.3-0.7g / L, K2HPO4 1-3g / L, pH 7;

[0090] The culture conditions were: temperature 35°C, shaker speed 250 r / min, inoculum size 8%, and culture time 30 h;

[0091] S3 according to the ratio of Table 1 methyl thioglycolate, dimethyl trisulfide, 3-methyl butyl sulfate S- methyl ester, 3-mercaptopropyltrimethoxysilane and water were mixed, and then the dry powder was added to obtain a mixture;

[0092] S4. The mixture was mixed with modified montmorillonite to obtain a composite mold removal agent preparation;

[0093] S5. Dry the composite mold removal agent and then package it.

[0094] Table 1 Raw material ratio table of Examples 1-8 (kg)

[0095] Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 Example 7 Example 8 Brevibacillus laterosporus 5 10 15 10 10 10 10 10 Methyl thioglycolate 25 20 10 20 20 20 20 20 dimethyl trisulfide 10 15 20 12 18 15 15 15 3-Methylbutane sulfate S-methyl ester 35 25 15 30 20 25 25 25 Modified montmorillonite 15 25 30 25 25 25 25 25 water 20 30 40 30 30 30 30 30 3-Mercaptopropyltrimethoxysilane 0 0 0 0 0 5 9 12

[0096] The modified montmorillonite comes from Preparation Example 1.

[0097] Example 9

[0098] The difference from Example 2 is that in Example 9, the amount of Brevibacillus laterosporus used is 7.5 kg.

[0099] Example 10

[0100] The difference from Example 2 is that in Example 10, the amount of Brevibacillus laterosporus used is 5 kg.

[0101] Examples 11-13

[0102] Different from Example 7, the modified montmorillonite in Examples 11-13 comes from Preparation Examples 2-4, respectively.

[0103] Examples 14-18

[0104] Different from Example 7, in Examples 14-18, the modified montmorillonite was replaced by an equal amount of hexadecyltrimethylammonium bromide-modified montmorillonite from Preparation Examples 5-9, respectively.

[0105] Comparative Example

[0106] Comparative Example 1

[0107] The difference from Example 1 is that in Comparative Example 1, an equal amount of montmorillonite is used to replace the modified montmorillonite.

[0108] Comparative Example 2

[0109] The difference from Example 3 is that in Comparative Example 2, 5 kg each of methyl thioglycolate, dimethyl trisulfide, and 3-methylbutyl sulfate S-methyl ester were used to replace 15 kg of Brevibacillus laterosporus.

[0110] Comparative Example 3

[0111] The difference from Example 1 is that in Comparative Example 3, an equal amount of methyl thioglycolate is used to replace dimethyl trisulfide.

[0112] Comparative Example 4

[0113] The difference from Example 1 is that in Comparative Example 4, 3-methylbutyrate S-methyl ester was replaced by an equal amount of dimethyl trisulfide.

[0114] Performance testing

[0115] 1. Mildew removal performance test

[0116] 7 g of the composite mold removal agent prepared in each Example and Comparative Example was added to 1 kg of contaminated corn flour (wherein the zearalenone content was 50 ppm, the aflatoxin B1 content was 50 ppm, and the vomitoxin content was 50 ppm, and the pH of the corn flour extract was 6). 1 kg of distilled water was added to each test group. Three replicates were performed for each group. After mixing thoroughly, the mixture was treated at 30°C for 24 hours. Samples from each test group were then accurately weighed, and the zearalenone, aflatoxin B1, and vomitoxin content of each sample was determined. The removal rate (%) was calculated according to the following formula. The results are shown in Table 2:

[0117] Removal rate (%) = (mass of mycotoxins in the sample before treatment - mass of mycotoxins in the sample after treatment) / mass of mycotoxins in the sample before treatment × 100%.

[0118] Table 2 Performance test results / %

[0119] Zearalenone removal rate Aflatoxin B1 removal rate Deoxynivalenol removal rate Example 1 96.6 96.1 96.4 Example 2 97.4 96.7 97.2 Example 3 96.4 95.9 96.2 Example 4 97.1 96.4 96.9 Example 5 96.9 96.2 96.7 Example 6 98.1 97.6 97.9 Example 7 98.4 97.9 98.2 Example 8 97.7 97.2 97.4 Example 9 98.2 97.5 97.8 Example 10 98.3 97.8 98.1 Example 11 98.4 97.5 97.8 Example 12 97.7 97.2 97.5 Example 13 97.1 96.6 96.9 Example 14 99.5 99.5 99.6 Example 15 99.2 99.2 99.3 Example 16 99.0 98.5 98.8 Example 17 99.2 99.0 99.1 Example 18 98.8 98.3 98.6 Comparative Example 1 83.8 84.2 82.6 Comparative Example 2 76.8 77.3 75.1 Comparative Example 3 87.8 88.6 86.8 Comparative Example 4 88.3 87.8 86.1

[0120] Combining Examples 1-18 with Comparative Examples 1-4 and Table 2, it can be seen that the composite de-mold agent obtained in Examples 1-18 has a higher removal rate of zearalenone, aflatoxin B1, and vomitoxin in corn flour than that in Comparative Examples 1-4, which indicates that the composite de-mold agent obtained in the present application has a better de-mold effect on corn flour. This may be because the modified montmorillonite in the composite de-mold agent of the present application adsorbs zearalenone, aflatoxin B1, and vomitoxin on corn flour, and also adsorbs part of Brevibacillus laterosporus, methyl thioglycolate, dimethyl trisulfide, and 3-methylbutane sulfate S-methyl ester, so that Brevibacillus laterosporus, methyl thioglycolate, dimethyl trisulfide, and 3-methylbutane sulfate S-methyl ester are in accurate contact with zearalenone, aflatoxin B1, and vomitoxin. Brevibacillus laterosporus, methyl thioglycolate, dimethyl trisulfide, and 3-methylbutane sulfate S-methyl ester cooperate with each other to rob zearalenone, aflatoxin B1, and vomitoxin of their living nutrients and living space, thereby inhibiting the growth of zearalenone, aflatoxin B1, and vomitoxin, and degrading zearalenone, aflatoxin B1, and vomitoxin, thereby achieving rapid and efficient de-mold.

[0121] Combining Example 1 with Comparative Example 1 and Table 2, it can be seen that the composite mold removal agent obtained in Example 1 has higher removal rates for zearalenone, aflatoxin B1, and vomitoxin from corn flour than that in Comparative Example 1. This indicates that the combination of modified montmorillonite with Brevibacillus laterosporus, methyl thioglycolate, dimethyl trisulfide, and 3-methylbutyl sulfate S-methyl ester helps improve the removal of zearalenone, aflatoxin B1, and vomitoxin. This may be because the copper ions grafted onto the montmorillonite not only improve the adsorption of mold by the montmorillonite, but also promote the removal of mold by Brevibacillus laterosporus, methyl thioglycolate, dimethyl trisulfide, and 3-methylbutyl sulfate S-methyl ester.

[0122] Combining Example 3 with Comparative Examples 2-4 and Table 2, it can be seen that the composite mold removal agent obtained in Example 3 has higher removal rates for zearalenone, aflatoxin B1, and vomitoxin in corn flour than those in Comparative Examples 2-4. This indicates that the combination of Brevibacillus laterosporus with methyl thioglycolate, dimethyl trisulfide, and 3-methylbutyrate S-methyl sulfate can further enhance mold removal. This may be because Brevibacillus laterosporus, methyl thioglycolate, dimethyl trisulfide, and 3-methylbutyrate S-methyl sulfate work synergistically. Methyl thioglycolate, dimethyl trisulfide, and 3-methylbutyrate S-methyl sulfate can rapidly eliminate mycotoxins. Simultaneously, methyl thioglycolate, dimethyl trisulfide, and 3-methylbutyrate S-methyl sulfate inhibit the release of substances from Brevibacillus laterosporus, resulting in a slower release of mold-removing substances from Brevibacillus laterosporus, extending the effective action time of the mold removal agent and enabling more thorough mold removal.

[0123] Combining Example 2 with Examples 6-8 and Table 2, it can be seen that the composite mold removal agents obtained in Examples 6-8 have higher removal rates for zearalenone, aflatoxin B1, and vomitoxin in corn flour than those in Example 2. This indicates that the addition of 3-mercaptopropyltrimethoxysilane can improve the mold removal effect. This may be because the physical mixing of 3-mercaptopropyltrimethoxysilane with montmorillonite enhances the colonization of probiotics on the montmorillonite through physical adsorption, thereby improving the mold removal effect. The addition amount of 3-mercaptopropyltrimethoxysilane in Examples 6-7 is more optimal.

[0124] Combining Example 7 with Examples 14-18 and Table 2, it can be seen that the composite de-mold agent obtained in Examples 14-18 has a higher removal rate of zearalenone, aflatoxin B1, and vomitoxin in corn flour than that in Example 7. This shows that grafting hexadecyltrimethylammonium bromide on the modified montmorillonite can further improve the de-mold effect. This may be because grafting hexadecyltrimethylammonium bromide increases the interlayer distance of montmorillonite, which on the one hand further improves the adsorption effect of montmorillonite, and on the other hand allows water molecules to better enter the interlayer of montmorillonite, thereby increasing the water solubility of montmorillonite and thus improving its de-mold effect.

[0125] 2. Toxicity Testing

[0126] Experimental animals: 70 clean-grade ICR Kunming mice (weight: 18±2g, half male and half female) aged 6-8 weeks were fasted for 12 hours but not deprived of water and randomly divided into control group and experimental group.

[0127] The control group was fed with 5g of mouse feed that was tested to be not contaminated by mold, for 15 consecutive days.

[0128] The experimental group was fed with the mouse feed treated with the mold removal agent in Example 7. The treatment method was as follows: the same mouse feed as the control group was ground into powder, treated with the mold removal agent at a weight ratio of mouse feed to mold removal agent of 300:1, and then dried and pressed into mouse feed. The daily feeding amount was 5g for 15 consecutive days.

[0129] Experimental results:

[0130] After one day of feeding: the mice in the control group and the experimental group had a normal diet, gained weight, moved freely, had smooth fur, had normal urination and defecation, no abnormal secretions, and no abnormal blood biochemical indicators.

[0131] After 5 days of feeding: the mice in the control group and the experimental group had a normal diet, gained weight, moved freely, had smooth fur, had normal urination and defecation, no abnormal secretions, and no abnormal blood biochemical indicators.

[0132] After 10 days of feeding: the mice in the control group and the experimental group had a normal diet, gained weight, moved freely, had smooth fur, had normal urination and defecation, no abnormal secretions, and no abnormal blood biochemical indicators.

[0133] After 15 days of feeding: the mice in the control group and the experimental group had a normal diet, gained weight, moved freely, had smooth fur, had normal urination and defecation, no abnormal secretions, and no abnormal blood biochemical indicators.

[0134] After the observation period, the mice were killed, and no obvious pathological changes were found in the mice in the control group and the experimental group through dissection.

[0135] This shows that the mold removal agent of the present application is non-toxic to mice and is safe and reliable.

[0136] Application Examples

[0137] Application Example 1

[0138] The composite mold removal agent obtained in Example 1 was applied to moldy feed, with the weight ratio of the moldy feed to the composite mold removal agent being 450:1.

[0139] Application Example 2-18

[0140] The difference from Application Example 1 is that the composite mildew removers in Application Examples 2-18 are respectively from Examples 2-18.

[0141] Application Example 19

[0142] Different from Application Example 1, the composite mold removal agent in Application Example 19 is applied to moldy corn flour.

[0143] This specific embodiment is merely an explanation of the present application and is not a limitation of the present application. After reading this specification, those skilled in the art may make non-creative modifications to the present embodiment as needed, but as long as they are within the scope of the claims of the present application, they are protected by the patent law.

Claims

1. A method for preparing a composite mold removal agent, characterized in that: The following steps are involved: S1. Preparation of modified montmorillonite: 1) Dissolve copper sulfate in water to prepare copper sulfate solution; 2) Add montmorillonite to the copper sulfate solution, and then add a pH adjuster to adjust the pH value of the solution to 10; 3) Add formaldehyde solution as a reducing agent and start the chemical deposition reaction at 70-75°C for 12-14 hours; 4) After the reaction is completed, filtering, washing, and drying to remove unreacted copper ions and residual reducing agent to obtain modified montmorillonite; S2. Brevibacillus laterosporus was cultured in a culture medium, and after completion of the culture, a Brevibacillus laterosporus bacterial solution was obtained, and then the resulting bacterial solution was prepared into a dry powder; S3 methyl thioglycolate, dimethyl trisulfide, 3-methylbutyl sulfate S- methyl ester was mixed with water, and then the dry powder was added to obtain a mixture; S4. The mixture was mixed with modified montmorillonite to obtain a composite mold removal agent preparation; S5. Dry the composite mold removal agent and then package it.

2. The method for preparing a composite mold removal agent according to claim 1, characterized in that: The concentration of the copper sulfate solution is 16-20 g / L; the weight ratio of copper sulfate to montmorillonite is 1:(5-12); and the weight ratio of formaldehyde to copper sulfate is (3-4):

5.

3. The method for preparing a composite mold removal agent according to claim 2, wherein: The weight ratio of the copper sulfate to the montmorillonite is 1:(6-8).

4. The method for preparing a composite mold removal agent according to claim 1, characterized in that: Hexadecyltrimethylammonium bromide is grafted onto the modified montmorillonite by the following grafting method: a) dissolving cetyltrimethylammonium bromide in water to obtain a cetyltrimethylammonium bromide solution, and adding methyl p-toluenesulfonate to the cetyltrimethylammonium bromide solution to obtain a treatment solution; b) adding the modified montmorillonite to the treatment solution, mixing, and reacting under condensation reflux at 70-85°C for 2-4 hours; c) After the reaction is completed, the slurry is ultrasonically dispersed, the supernatant is removed by centrifugation, the precipitate is collected, and the precipitate is washed; the washed precipitate is dried at 80-90° C. to obtain hexadecyltrimethylammonium bromide-modified montmorillonite.

5. The method for preparing a composite mold removal agent according to claim 4, characterized in that: The weight ratio of hexadecyltrimethyl bromide to methyl p-toluenesulfonate is 1:(0.1-0.3); the weight ratio of hexadecyltrimethyl bromide to modified montmorillonite is 1:(35-42).

6. The method for preparing a composite mold removal agent according to claim 1, characterized in that: In S3, methyl thioglycolate, dimethyl trisulfide, 3-methylbutyl sulfate S-methyl ester, and 3-mercaptopropyltrimethoxysilane are mixed with water, and then the dry powder is added to obtain a mixed solution.

7. A composite mold removal agent prepared by the preparation method of the composite mold removal agent according to any one of claims 1 to 6, characterized in that: The invention comprises the following raw materials in parts by weight: 5-15 parts of Brevibacillus laterosporus, 15-25 parts of methyl thioglycolate, 10-20 parts of dimethyl trisulfide, 15-35 parts of 3-methylbutyl sulfate S-methyl ester, 15-30 parts of modified montmorillonite, 20-40 parts of water and 5-9 parts of optional 3-mercaptopropyltrimethoxysilane.

8. The composite mold removal agent according to claim 7, characterized in that: The weight ratio of the methyl thioglycolate, dimethyl trisulfide and 3-methylbutyl sulfate S-methyl ester is 1: (0.6-0.9): (1-1.5).

9. The composite mold removal agent according to claim 7, characterized in that: The weight ratio of the sum of the weights of the methyl thioglycolate, dimethyl trisulfide, and 3-methylbutyl sulfate S-methyl ester to the weight of Brevibacillus laterosporus is (8-12):

1.

10. The composite mold removal agent according to claim 7, characterized in that: Applied to moldy grains or feed, the weight ratio of moldy grains or feed to compound mold removal agent is (400-500):1.