A complex enzyme preparation for degrading mycotoxins in feed and a preparation method and application thereof

Abstract

The present application belongs to the field of feed resources, and particularly relates to a compound enzyme preparation for degrading mycotoxins in feed, a preparation method and application thereof. The compound enzyme preparation comprises the following components in parts by weight: mycotoxin-degrading enzyme 25-35 parts, compound probiotic powder 15-30 parts, Chrysanthemum morifolium Ramat extract 10-25 parts, Cynanchum thesioides Hemsl extract 8-12 parts, and montmorillonite 6-20 parts. The compound enzyme preparation for degrading mycotoxins in feed is added to feed, which can significantly reduce the number of molds in feed, improve daily weight gain, reduce feed conversion ratio, and effectively promote the growth and development ability of animals. The compound enzyme preparation for degrading mycotoxins in feed is green and non-toxic, meets food safety, and has a broad application prospect.

Description

Technical Field

[0001] This invention belongs to the field of feed resources, specifically relating to a compound enzyme preparation for degrading mycotoxins in feed, its preparation method, and its application. Background Technology

[0002] Mycotoxins are toxic metabolites produced by molds in contaminated food. Fungi can produce mycotoxins under suitable conditions, whether during grain growth, feed manufacturing, storage, or transportation. In recent years, the frequency of mycotoxin contamination in feed ingredients has been increasing globally. Currently, more than 200 types of mycotoxins have been found in feed, with common ones including aflatoxin, zearalenone, vomitoxin, and trichothecotoxins. Mycotoxins typically reduce animal production performance, suppress immune function, cause disease, necrotic and hemorrhagic dermatitis, decrease feed intake, inhibit growth, and result in poor daily weight gain. They then enter the human food chain through animal meat, milk, and offal, harming human health and causing significant economic losses to the livestock industry. Therefore, inhibiting mycotoxins in feed is crucial for solving mold contamination and protecting animal health.

[0003] Currently, the main methods for mycotoxin detoxification applied in the feed industry are physical adsorption and biodegradation. Physical adsorption involves adding mycotoxin adsorbents to the feed. This method is simple to operate and relatively low-cost; however, while adsorbing toxins, it also adsorbs vitamins, minerals, and other nutrients in the feed, reducing its nutritional value. Furthermore, it is not very effective at adsorbing some polar toxins. Biodegradation utilizes mycotoxin-degrading enzymes to break down the structure of mycotoxins, converting them into non-toxic products, thus achieving detoxification. Biodegradation detoxification can completely decompose toxins, leaving no toxic residues. It targets only mycotoxins without damaging other components in the feed and raw materials, does not reduce the nutritional value of the feed, and does not alter the physical and sensory composition of the product, meeting the requirements of green and safe practices.

[0004] To achieve the above objectives, the present invention provides a compound enzyme preparation for degrading mycotoxins in feed, its preparation method, and its application. Summary of the Invention

[0005] The primary objective of this invention is to provide a compound enzyme preparation for degrading mycotoxins in feed.

[0006] The second objective of this invention is to provide a method for preparing a compound enzyme preparation that degrades mycotoxins in feed.

[0007] A third objective of this invention is to provide the application of a complex enzyme preparation for degrading mycotoxins in feed in the preparation of feed additives.

[0008] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0009] A compound enzyme preparation for degrading mycotoxins in feed, the compound enzyme preparation comprising the following components in parts by weight: 25-35 parts mycotoxin degrading enzyme, 15-30 parts compound probiotic powder, 10-25 parts *Citrus medica* extract, 8-12 parts *Aristolochia debilis* extract, and 6-20 parts montmorillonite.

[0010] Preferably, the specific preparation method of the compound probiotic powder is as follows: Bacillus subtilis and Lactobacillus plantarum are activated separately to obtain Bacillus subtilis bacterial solution and Lactobacillus plantarum bacterial solution respectively; the Bacillus subtilis bacterial solution and Lactobacillus plantarum bacterial solution are inoculated into a fermentation medium and cultured at 35-40℃ for 30-40h to obtain fermentation product; the fermentation product is freeze-dried to obtain compound probiotic powder.

[0011] Preferably, the fermentation medium comprises: glucose 16-21 g / L, peptone 8-11 g / L, ammonium nitrate 2.0-4.0 g / L, tyrosine 3.0-5.6 g / L, L-glutamyl dipeptide 0.2-0.9 g / L, yeast extract 3.5-5.0 g / L, magnesium sulfate 0.3-0.6 g / L, zinc sulfate 0.1-0.23 g / L, ferric sulfate 0.1-0.3 g / L, manganese sulfate 0.2-0.25 g / L, potassium dihydrogen phosphate 3.0-4.2 g / L, and Tween-80 0.5-2.0 g / L.

[0012] Preferably, the specific steps for activating Bacillus subtilis are as follows: Bacillus subtilis is inoculated into LB medium and cultured at 35-40℃ for 24-28 hours to obtain Bacillus subtilis bacterial solution; the specific steps for activating Lactobacillus plantarum are as follows: Lactobacillus plantarum is inoculated into MRS medium and cultured at 35-40℃ for 18-24 hours to obtain Lactobacillus plantarum bacterial solution.

[0013] Preferably, the effective viable count of the Bacillus subtilis bacterial solution is (1-3)×10⁻⁶. 8 CFU / mL; the effective viable count of the *Lactobacillus plantarum* bacterial suspension was (1-5)×10⁻⁶. 8 CFU / mL; the Bacillus subtilis culture, Lactobacillus plantarum culture and fermentation medium were inoculated at a volume ratio of 5:(5-7):100.

[0014] Preferably, the specific preparation method of the *Chrysanthemum indicum* extract is as follows:

[0015] (1) Fresh chrysanthemum is washed, dried and crushed to obtain chrysanthemum powder;

[0016] (2) Add the chrysanthemum powder to an ethanol solution with a volume concentration of 65-80%, the amount of ethanol solution added is 10-15 times the mass of the chrysanthemum powder, heat at 30-45℃ for 2-3 hours; filter, centrifuge, and collect the first filtrate and the first filter residue;

[0017] (3) Then add an ethanol solution with a volume concentration of 65-80% to the first filter residue. The amount of ethanol solution added is 10-15 times the mass of the first filter residue. Heat at 30-45℃ for 2-3 hours. After filtration and centrifugation, collect the second filtrate. Combine the first and second filtrates, concentrate and freeze dry to obtain the chrysanthemum extract.

[0018] Preferably, the specific preparation method of the Akebia quinata extract is as follows:

[0019] (1) Take Tibetan akebia, wash it, dry it, and crush it to make Tibetan akebia powder;

[0020] (2) Soak the Tibetan Akebia powder in 10-15 times its weight of water for 2-3 hours, then decoct for 3-4 hours; filter, centrifuge, and collect the first filtrate and the first residue;

[0021] (3) Then add 10-15 times the weight of water to the first filter residue and soak for 2-3 hours, then decoct for 2-3 hours; after filtration and centrifugation, collect the second filtrate, combine the first and second filtrates, concentrate and freeze dry to obtain the Tibetan Akebia extract.

[0022] Preferably, the mycotoxin degrading enzyme comprises, by weight, 1-2 parts aflatoxin degrading enzyme, 2-6 parts zearalenone degrading enzyme, and 1-5 parts vomitoxin degrading enzyme.

[0023] According to the preparation method of the compound enzyme preparation for degrading mycotoxins in feed described above, the preparation method is as follows: Weigh the following components according to the specified weight proportions: mycotoxin degrading enzyme, compound probiotic powder, chrysanthemum extract, akebia quinata extract, and montmorillonite, and stir evenly at room temperature to obtain the final product.

[0024] The above describes the application of compound enzyme preparations for degrading mycotoxins in feed in the preparation of feed additives.

[0025] Compared with the prior art, the main advantages of the present invention are as follows:

[0026] This invention provides a compound enzyme preparation for degrading mycotoxins in feed, comprising a mycotoxin-degrading enzyme, a compound probiotic powder, *Cynanchum paniculatum* extract, *Akebia quinata* extract, and montmorillonite. *Cynanchum paniculatum* extract and *Akebia quinata* extract exhibit synergistic effects, effectively inhibiting mold growth. Furthermore, the addition of L-glutamyl dipeptide to the probiotic fermentation medium enhances the mycotoxin degradation efficiency. Experimental results show that adding this compound enzyme preparation to feed significantly reduces the amount of mold, increases daily weight gain, lowers the feed conversion ratio, and effectively promotes animal growth and development. Moreover, this compound enzyme preparation for degrading mycotoxins in feed is green and non-toxic, meets food safety standards, and has broad application prospects. Detailed Implementation

[0027] The technical solution of the present invention will be further described below with reference to specific embodiments. However, those skilled in the art should understand that the following embodiments are only for illustrating the present invention and should not be regarded as limiting the present invention. Specific conditions not specified in the embodiments are performed according to conventional conditions or conditions recommended by the manufacturer. Unless otherwise specified, the reagents or instruments used are all conventional products obtained through commercial channels.

[0028] The preparation method of the *Chrysanthemum indicum* extract used in this embodiment of the invention is as follows:

[0029] (1) Fresh chrysanthemum is washed, dried and crushed to obtain chrysanthemum powder;

[0030] (2) Add the chrysanthemum powder to an ethanol solution with a volume concentration of 75%, the amount of ethanol solution added is 12 times the mass of the chrysanthemum powder, heat at 37°C for 2 hours; filter, centrifuge, and collect the first filtrate and the first filter residue;

[0031] (3) Then add a 75% ethanol solution to the first filter residue. The amount of ethanol solution added is 12 times the mass of the first filter residue. Heat at 37°C for 2 hours to extract. After filtration and centrifugation, collect the second filtrate. Combine the first and second filtrates, concentrate and freeze dry to obtain the chrysanthemum extract.

[0032] The preparation method of the Akebia quinata extract used in this embodiment of the invention is as follows:

[0033] (1) Take Tibetan akebia, wash it, dry it, and crush it to make Tibetan akebia powder;

[0034] (2) Soak the Tibetan akebia powder in 12 times its weight of water for 2 hours, then decoct for 3 hours; filter, centrifuge, and collect the first filtrate and the first residue;

[0035] (3) Then add 12 times the weight of water to the first filter residue and soak for 2 hours, then decoct for 2 hours; after filtration and centrifugation, collect the second filtrate, combine the first and second filtrates, concentrate and freeze dry to obtain the extract of Aristolochia debilis.

[0036] Example 1

[0037] A compound enzyme preparation for degrading mycotoxins in feed, the compound enzyme preparation comprising the following components in parts by weight: 30 parts mycotoxin degrading enzyme, 22 parts compound probiotic powder, 17 parts *Citrus aurantium* extract, 10 parts *Aristolochia debilis* extract, and 13 parts montmorillonite; wherein the mycotoxin degrading enzyme, by weight, comprises 1 part aflatoxin degrading enzyme, 4 parts zearalenone degrading enzyme, and 3 parts vomitoxin degrading enzyme.

[0038] The specific preparation method of the compound probiotic powder is as follows:

[0039] (1) Bacillus subtilis was inoculated into 2 mL of LB medium and cultured at 37°C for 26 h to obtain Bacillus subtilis bacterial suspension; Lactobacillus plantarum was inoculated into 2 mL of MRS medium and cultured at 37°C for 21 h to obtain Lactobacillus plantarum bacterial suspension; the effective viable count of the Bacillus subtilis bacterial suspension was 2 × 10⁻⁶. 8 CFU / mL; the effective viable count of the *Lactobacillus plantarum* bacterial suspension was 3 × 10⁻⁶. 8 CFU / mL.

[0040] (2) Dissolve glucose, peptone, ammonium nitrate, tyrosine, L-glutamyl dipeptide, yeast extract, magnesium sulfate, zinc sulfate, ferric sulfate, manganese sulfate, potassium dihydrogen phosphate, and Tween-80 in deionized water according to their respective concentrations and usage amounts. Mix well, adjust the pH to 6.0, and sterilize at 115℃ for 20 min to obtain the fermentation medium. The final concentrations of the fermentation medium are as follows: glucose 18 g / L, peptone 9 g / L, ammonium nitrate 3.0 g / L, tyrosine 4.3 g / L, L-glutamyl dipeptide 0.5 g / L, yeast extract 4.2 g / L, magnesium sulfate 0.4 g / L, zinc sulfate 0.16 g / L, ferric sulfate 0.2 g / L, manganese sulfate 0.22 g / L, potassium dihydrogen phosphate 3.6 g / L, and Tween-80 1.2 g / L.

[0041] (3) Inoculate the Bacillus subtilis culture, Lactobacillus plantarum culture and the fermentation medium prepared in step (2) at a volume ratio of 5:6:100, and incubate at 37°C for 35 hours to obtain the fermentation product; freeze-dry the fermentation product to obtain the compound probiotic powder.

[0042] This embodiment also provides a method for preparing a compound enzyme preparation for degrading mycotoxins in feed. The specific preparation method is as follows:

[0043] Weigh out the mycotoxin degrading enzyme, compound probiotic powder, chrysanthemum extract, akebia quinata extract, and montmorillonite according to the stated weight proportions, and stir evenly at room temperature to obtain the final product.

[0044] Example 2

[0045] A compound enzyme preparation for degrading mycotoxins in feed, the compound enzyme preparation comprising the following components in parts by weight: 25 parts mycotoxin degrading enzyme, 15 parts compound probiotic powder, 10 parts *Citrus aurantium* extract, 8 parts *Aristolochia debilis* extract, and 6 parts montmorillonite; wherein the mycotoxin degrading enzyme, by weight, comprises 1 part aflatoxin degrading enzyme, 2 parts zearalenone degrading enzyme, and 1 part vomitoxin degrading enzyme.

[0046] The specific preparation method of the compound probiotic powder is as follows:

[0047] (1) Bacillus subtilis was inoculated into 2 mL of LB medium and cultured at 37°C for 24 h to obtain Bacillus subtilis bacterial suspension; Lactobacillus plantarum was inoculated into 2 mL of MRS medium and cultured at 37°C for 18 h to obtain Lactobacillus plantarum bacterial suspension; the effective viable count of the Bacillus subtilis bacterial suspension was 1 × 10⁻⁶. 8 CFU / mL; the effective viable count of the *Lactobacillus plantarum* bacterial suspension is 1×10⁻⁶. 8 CFU / mL.

[0048] (2) Dissolve glucose, peptone, ammonium nitrate, tyrosine, L-glutamyl dipeptide, yeast extract, magnesium sulfate, zinc sulfate, ferric sulfate, manganese sulfate, potassium dihydrogen phosphate, and Tween-80 in deionized water according to their respective concentrations and usage amounts. Mix well, adjust the pH to 6.0, and sterilize at 115℃ for 20 min to obtain the fermentation medium. The final concentrations of the fermentation medium are as follows: glucose 16 g / L, peptone 8 g / L, ammonium nitrate 2.0 g / L, tyrosine 3.0 g / L, L-glutamyl dipeptide 0.2 g / L, yeast extract 3.5 g / L, magnesium sulfate 0.3 g / L, zinc sulfate 0.1 g / L, ferric sulfate 0.1 g / L, manganese sulfate 0.2 g / L, potassium dihydrogen phosphate 3.0 g / L, and Tween-80 0.5 g / L.

[0049] (3) Inoculate the Bacillus subtilis culture, Lactobacillus plantarum culture and the fermentation medium prepared in step (2) at a volume ratio of 5:5:100, and incubate at 37°C for 30 h to obtain the fermentation product; freeze-dry the fermentation product to obtain the compound probiotic powder.

[0050] This embodiment also provides a method for preparing a compound enzyme preparation for degrading mycotoxins in feed. The specific preparation method is as follows:

[0051] Weigh out the mycotoxin degrading enzyme, compound probiotic powder, chrysanthemum extract, akebia quinata extract, and montmorillonite according to the stated weight proportions, and stir evenly at room temperature to obtain the final product.

[0052] Example 3

[0053] A compound enzyme preparation for degrading mycotoxins in feed, the compound enzyme preparation comprising the following components in parts by weight: 35 parts mycotoxin degrading enzyme, 30 parts compound probiotic powder, 25 parts *Citrus aurantium* extract, 12 parts *Aristolochia debilis* extract, and 20 parts montmorillonite; wherein the mycotoxin degrading enzyme, by weight, comprises 2 parts aflatoxin degrading enzyme, 6 parts zearalenone degrading enzyme, and 5 parts vomitoxin degrading enzyme.

[0054] The specific preparation method of the compound probiotic powder is as follows:

[0055] (1) Bacillus subtilis was inoculated into 2 mL of LB medium and cultured at 37°C for 28 h to obtain Bacillus subtilis bacterial suspension; Lactobacillus plantarum was inoculated into 2 mL of MRS medium and cultured at 37°C for 24 h to obtain Lactobacillus plantarum bacterial suspension; the effective viable count of the Bacillus subtilis bacterial suspension was 3 × 10⁻⁶. 8 CFU / mL; the effective viable count of the *Lactobacillus plantarum* bacterial suspension was 5 × 10⁻⁶. 8 CFU / mL.

[0056] (2) Dissolve glucose, peptone, ammonium nitrate, tyrosine, L-glutamyl dipeptide, yeast extract, magnesium sulfate, zinc sulfate, ferric sulfate, manganese sulfate, potassium dihydrogen phosphate, and Tween-80 in deionized water according to their respective concentrations and usage amounts. Mix well, adjust the pH to 6.0, and sterilize at 115℃ for 20 min to obtain the fermentation medium. The final concentrations of the fermentation medium are as follows: glucose 21 g / L, peptone 11 g / L, ammonium nitrate 4.0 g / L, tyrosine 5.6 g / L, L-glutamyl dipeptide 0.9 g / L, yeast extract 5.0 g / L, magnesium sulfate 0.6 g / L, zinc sulfate 0.23 g / L, ferric sulfate 0.3 g / L, manganese sulfate 0.25 g / L, potassium dihydrogen phosphate 4.2 g / L, and Tween-80 2.0 g / L.

[0057] (3) Inoculate the Bacillus subtilis culture, Lactobacillus plantarum culture and the fermentation medium prepared in step (2) at a volume ratio of 5:7:100, and incubate at 37℃ for 40h to obtain the fermentation product; freeze-dry the fermentation product to obtain the compound probiotic powder.

[0058] This embodiment also provides a method for preparing a compound enzyme preparation for degrading mycotoxins in feed. The specific preparation method is as follows:

[0059] Weigh out the mycotoxin degrading enzyme, compound probiotic powder, chrysanthemum extract, akebia quinata extract, and montmorillonite according to the stated weight proportions, and stir evenly at room temperature to obtain the final product.

[0060] Comparative Example 1

[0061] The difference between Comparative Example 1 and Example 1 is that the compound enzyme preparation for degrading mycotoxins in feed omits the extract of *Gynostemma pentaphyllum*, while all other components are the same as in Example 1.

[0062] Comparative Example 2

[0063] The difference between Comparative Example 2 and Example 1 is that the extract of Aristolochia debilis was omitted from the complex enzyme preparation for degrading mycotoxins in feed, while all other components are the same as in Example 1.

[0064] Comparative Example 3

[0065] The difference between Comparative Example 3 and Example 1 is that the extract of *Gynostemma pentaphyllum* in the compound enzyme preparation for degrading mycotoxins in feed was replaced with extract of *Aristolochia debilis*. All other aspects are the same as in Example 1.

[0066] Comparative Example 4

[0067] The difference between Comparative Example 4 and Example 1 is that the extract of Aristolochia debilis in the compound enzyme preparation for degrading mycotoxins in feed is replaced with extract of Chrysanthemum indicum. All other aspects are the same as in Example 1.

[0068] Comparative Example 5

[0069] The difference between Comparative Example 5 and Example 1 is that L-glutamyl dipeptide was omitted from the fermentation medium prepared in step (2), while the rest were the same as in Example 1.

[0070] Experimental Example 1

[0071] The effect of mycotoxin degradation complex enzyme preparations on mold count:

[0072] 9000g of fattening pig feed was weighed and divided into four groups: control group, Example 1 group, Example 2 group, Example 3 group, Comparative Example 1 group, Comparative Example 2 group, Comparative Example 3 group, Comparative Example 4 group, and Comparative Example 5 group, with 1000g in each group. 10g of the compound enzyme preparation prepared in Examples 1-3 and Comparative Examples 1-5 was added to each group, while no product was added to the control group. After mixing, the mixture was bagged and placed in a test chamber at 30℃ and 75% relative humidity. Starting from day 20, the growth of mold was observed and recorded daily. Samples were taken after day 50, and the total mold count in each group was determined according to "GB / T13092-2006 Determination of Total Mold Count in Feed". The results of the total mold count in each group were recorded, as shown in Table 1.

[0073] Table 1. Number of molds in each group of feeds after 50 days of storage following the addition of compound enzyme preparation.

[0074]

[0075]

[0076] The results are shown in Table 1, which shows the number of molds in each group of feed after 50 days of storage following the addition of the compound enzyme preparation. As can be seen from the table, by adding the compound enzyme preparation to the feed, compared with comparative groups 1-5, the compound enzyme preparations prepared in Examples 1-3 of this invention can significantly reduce the number of molds in the feed. This indicates that the combination of mycotoxin-degrading enzymes, compound probiotic powder, *Gynostemma pentaphyllum* extract, *Akebia quinata* extract, and montmorillonite in the compound enzyme preparation prepared in this invention can significantly reduce the number of molds in the feed.

[0077] Experimental Example 2

[0078] Effects of mycotoxin degradation complex enzyme preparations on pig growth performance:

[0079] This experiment selected 90 fattening pigs weighing approximately 50 kg each and randomly divided them into a control group, Example 1 group, Example 2 group, Example 3 group, Comparative Example 1 group, Comparative Example 2 group, Comparative Example 3 group, Comparative Example 4 group, and Comparative Example 5 group, with 10 pigs in each group. After routine immunization, they entered the experimental period. Example 1 group: 1% of the compound enzyme preparation prepared in Example 1 was added to the feed. The difference between the control group and Example 1 group was that the compound enzyme preparation prepared in Example 1 was not added to the feed during the feeding process; the feeding and other contents were the same as those in Example 1 group. The other groups added 1% of the compound enzyme preparation corresponding to their group to the fattening pig feed. All groups were fed continuously for 60 days. The fattening pig feed formula, by weight percentage, included the following raw materials: corn 65%, wheat bran 13%, soybean meal 15%, dicalcium phosphate 1%, calcium carbonate 2%, salt 2%, and fattening pig compound premix 2%. All groups had the same feed weight, pen conditions, and management methods. They were fed four times a day with free access to food and water. The starting weight, ending weight, daily weight gain, and feed consumption were recorded. The average starting weight, average ending weight, average daily weight gain, and feed conversion ratio for each group were also calculated. The results are shown in Table 2.

[0080] Table 2 Key indicators of fattening effect of pigs in each group

[0081]

[0082]

[0083] The results are shown in Table 2, which represents key indicators of the fattening effect of pigs in each group. As can be seen from the table, by adding the compound enzyme preparation to the feed, compared with comparative groups 1-5, the compound enzyme preparations prepared in Examples 1-3 of this invention can significantly improve the daily weight gain of pigs, reduce the feed conversion ratio, and effectively promote the growth and development of pigs. This indicates that the mycotoxin-degrading compound enzyme preparations prepared in Examples 1-3 of this invention can effectively prevent the formation of mycotoxins in feed, reduce the impact of mycotoxins on pigs, save feed costs, and improve economic efficiency.

[0084] Further analysis revealed that, compared to Example 1, the compound enzyme preparation prepared in Comparative Example 1 omitted *Corydalis yanhusuo* extract; the compound enzyme preparation prepared in Comparative Example 2 omitted *Akebia quinata* extract; the compound enzyme preparation prepared in Comparative Example 3 omitted *Corydalis yanhusuo* extract, and the amount of *Akebia quinata* extract was adjusted to the sum of the two; the compound enzyme preparation prepared in Comparative Example 4 omitted *Akebia quinata* extract, and the amount of *Corydalis yanhusuo* extract was adjusted to the sum of the two; and the fermentation medium composition in Comparative Example 5 omitted L-glutamyl dipeptide. The effectiveness of the compound enzyme preparations in degrading mycotoxins in all five groups was significantly reduced. This indicates that *Corydalis yanhusuo* extract and *Akebia quinata* extract in the compound enzyme preparation have a synergistic effect; and the addition of L-glutamyl dipeptide to the fermentation medium improved the degradation efficiency of mycotoxins; their combined use effectively improves the stability of the compound enzyme preparation, maintains its good activity, and thus effectively prevents the formation of mycotoxins in feed.

[0085] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them. The basic principles and main features of the present invention have been described above with specific implementation schemes. Based on the present invention, some modifications or substitutions can be made, but these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of protection claimed by the present invention.

Claims

1. A complex enzyme preparation for degrading mycotoxins in feed, characterized in that, The complex enzyme preparation comprises the following components in weight parts: mycotoxin-degrading enzyme 25-35 parts, complex probiotic powder 15-30 parts, gongju extract 10-25 parts, clematis extract 8-12 parts, and montmorillonite 6-20 parts. The specific preparation method of the gongju extract is as follows: (1) fresh gongju is washed, dried, and crushed to obtain gongju powder; (2) the gongju powder is added into an ethanol solution with a volume concentration of 65-80%, and the added amount of the ethanol solution is 10-15 times the mass of the gongju powder; the mixture is heated at 30-45℃ for 2-3h; the first filtrate and the first filter residue are collected after filtration and centrifugation; (3) then, the first filter residue is added into an ethanol solution with a volume concentration of 65-80%, and the added amount of the ethanol solution is 10-15 times the mass of the first filter residue; the mixture is heated at 30-45℃ for 2-3h; the second filtrate is collected after filtration and centrifugation; the first filtrate and the second filtrate are combined; and the combined filtrate is concentrated and freeze-dried to obtain the gongju extract. The specific preparation method of the clematis extract is as follows: (1) clematis is washed, dried, and crushed to obtain clematis powder; (2) the clematis powder is soaked in 10-15 times its weight of water for 2-3h, and then boiled for 3-4h; the first filtrate and the first filter residue are collected after filtration and centrifugation; (3) then, the first filter residue is soaked in 10-15 times its weight of water for 2-3h, and then boiled for 2-3h; the second filtrate is collected after filtration and centrifugation; the first filtrate and the second filtrate are combined; and the combined filtrate is concentrated and freeze-dried to obtain the clematis extract.

2. The complex enzyme preparation for degrading mycotoxins in feed according to claim 1, characterized by, The specific preparation method of the complex probiotic powder is as follows: Bacillus subtilis and Lactobacillus plantarum are activated respectively to obtain Bacillus subtilis bacterial liquid and Lactobacillus plantarum bacterial liquid respectively; the Bacillus subtilis bacterial liquid and the Lactobacillus plantarum bacterial liquid are inoculated into a fermentation medium and cultured at 35-40℃ for 30-40h to obtain a fermentation product; and the fermentation product is freeze-dried to obtain the complex probiotic powder.

3. The complex enzyme preparation for degrading mycotoxins in feed according to claim 2, characterized by, The fermentation medium comprises the following components: glucose 16-21g / L, peptone 8-11g / L, ammonium nitrate 2.0-4.0g / L, tyrosine 3.0-5.6g / L, L-alanyl-L-glutamine 0.2-0.9g / L, yeast extract 3.5-5.0g / L, magnesium sulfate 0.3-0.6g / L, zinc sulfate 0.1-0.23g / L, ferrous sulfate 0.1-0.3g / L, manganese sulfate 0.2-0.25g / L, potassium dihydrogen phosphate 3.0-4.2g / L, and Tween-80 0.5-2.0g / L.

4. The complex enzyme preparation for degrading mycotoxins in feed according to claim 2, characterized by, The specific activation steps of the Bacillus subtilis are as follows: the Bacillus subtilis is inoculated into LB medium and cultured at 35-40℃ for 24-28h to obtain Bacillus subtilis bacterial liquid; and the specific activation steps of the Lactobacillus plantarum are as follows: the Lactobacillus plantarum is inoculated into MRS medium and cultured at 35-40℃ for 18-24h to obtain Lactobacillus plantarum bacterial liquid.

5. The complex enzyme preparation for degrading mycotoxins in feed according to claim 2, characterized by, The effective viable cell number of the bacillus subtilis liquid is (1-3) × 10 8 CFU / mL; the effective viable cell number of the lactobacillus plantarum liquid is (1-5) × 10 8 CFU / mL; the bacillus subtilis liquid, the lactobacillus plantarum liquid and the fermentation medium are inoculated at a volume ratio of 5:(5-7):

100.

6. The complex enzyme preparation for degrading mycotoxins in feed according to claim 1, characterized by, The mycotoxin-degrading enzyme comprises, in weight parts, 1-2 parts of aflatoxin-degrading enzyme, 2-6 parts of zearalenone-degrading enzyme, and 1-5 parts of vomitoxin-degrading enzyme.

7. The method of claim 1-6, wherein the method is characterized in that, The preparation method is as follows: according to the weight parts, mycotoxin degrading enzyme, compound probiotic powder, chrysanthemum morifolium ramat extract, clematis chinensis osb extract and montmorillonite are taken and stirred uniformly at room temperature, and the product is obtained.

8. Use of a complex enzyme preparation degrading mycotoxins in feed according to any one of claims 1 to 6 for the preparation of a feed additive.

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