Complex enzyme preparation for degrading mycotoxin in feed as well as preparation method and application of complex enzyme preparation

Abstract

The invention belongs to the field of feed resources, and particularly relates to a compound enzyme preparation for degrading mycotoxin in feed as well as a preparation method and application of the compound enzyme preparation. The compound enzyme preparation is prepared from the following components in parts by weight: 25 to 35 parts of mycotoxin degrading enzyme, 15 to 30 parts of compound probiotic powder, 10 to 25 parts of flos chrysanthemi extract, 8 to 12 parts of caulis clematidis armandii extract and 6 to 20 parts of montmorillonite. When the compound enzyme preparation for degrading mycotoxin in the feed is added into the feed, the number of mould in the feed can be remarkably reduced, the daily gain can be increased, the feed-gain ratio can be reduced, and the growth and development capability of animals can be effectively promoted. And the compound enzyme preparation for degrading mycotoxin in the feed is green and non-toxic, accords with food safety, and has a wide application prospect.

Description

Technical Field

[0001] The present invention belongs to the field of feed resources, and in particular relates to a composite enzyme preparation for degrading mycotoxins in feed, and a preparation method and application thereof. Background Art

[0002] Mycotoxins primarily refer to toxic metabolites produced by mold in contaminated foods. Fungi can produce mycotoxins under the right conditions, whether during the growing of grains in the field or during feed manufacturing, storage, and transportation. In recent years, the frequency of mycotoxin contamination in feed ingredients has increased annually worldwide. Currently, over 200 mycotoxins have been found in feed, including aflatoxins, zearalenone, vomitoxin, and trichothecenes. Mycotoxins often reduce animal performance, suppress immune function, and cause illness, including necrotizing and hemorrhagic dermatitis, decreased feed intake, growth inhibition, and poor daily gain. Mycotoxins then enter the human food chain through animal meat, milk, and offal, endangering human health and causing significant economic losses to industries such as animal husbandry. Therefore, controlling mycotoxins in feed is key to addressing mold contamination and protecting animal health.

[0003] The main mycotoxin detoxification methods currently used in the feed industry include physical adsorption and biodegradation. Physical adsorption involves adding a mycotoxin adsorbent to the feed. This method is simple to operate and relatively low in cost. However, while adsorbing the toxins, it also adsorbs vitamins, minerals, and other nutrients in the feed, reducing the nutritional value of the feed. It also has poor adsorption effects on some polar toxins. Biodegradation utilizes mycotoxin-degrading enzymes to destroy the structure of mycotoxins, converting them into non-toxic products and achieving detoxification. Biodegradation can completely decompose toxins, leaving no toxic residues. It only targets mycotoxins without damaging other components in the feed or raw materials, reducing the nutritional value of the feed or changing the physical and sensory properties of the product, thus meeting green and safety requirements.

[0004] Based on the above objectives, the present invention provides a composite enzyme preparation for degrading mycotoxins in feed, as well as a preparation method and application thereof. Summary of the Invention

[0005] The first object of the present invention is to provide a composite enzyme preparation for degrading mycotoxins in feed.

[0006] The second object of the present invention is to provide a method for preparing a composite enzyme preparation for degrading mycotoxins in feed.

[0007] The third object of the present invention is to provide a composite enzyme preparation for degrading mycotoxins in feed for use in preparing feed additives.

[0008] In order to achieve the above object, the technical solution adopted by the present invention is:

[0009] A composite enzyme preparation for degrading mycotoxins in feed, comprising the following components in parts by weight: 25-35 parts of mycotoxin degrading enzyme, 15-30 parts of composite probiotic powder, 10-25 parts of chrysanthemum morifolium extract, 8-12 parts of Tibetan clematis extract, and 6-20 parts of montmorillonite.

[0010] Preferably, the specific preparation method of the composite probiotic powder is as follows: after activating Bacillus subtilis and Lactobacillus plantarum respectively, Bacillus subtilis bacterial liquid and Lactobacillus plantarum bacterial liquid are obtained respectively; the Bacillus subtilis bacterial liquid and the Lactobacillus plantarum bacterial liquid are inoculated into a fermentation medium and cultured at 35-40° C. for 30-40 hours to obtain a fermentation product; and the fermentation product is freeze-dried to obtain the composite probiotic powder.

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

[0012] Preferably, the specific steps of activating Bacillus subtilis are: inoculating Bacillus subtilis into LB medium, culturing at 35-40° C. for 24-28 hours, and obtaining a Bacillus subtilis bacterial liquid; the specific steps of activating Lactobacillus plantarum are: inoculating Lactobacillus plantarum into MRS medium, culturing at 35-40° C. for 18-24 hours, and obtaining a Lactobacillus plantarum bacterial liquid.

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

[0014] Preferably, the specific preparation method of the chrysanthemum extract is as follows:

[0015] (1) Wash, dry, and crush fresh chrysanthemum to obtain chrysanthemum powder;

[0016] (2) adding the chrysanthemum powder to an ethanol solution having a volume concentration of 65-80%, wherein the amount of the ethanol solution added is 10-15 times the mass of the chrysanthemum powder, and heating and extracting at 30-45° C. for 2-3 hours; filtering and centrifuging to collect a first filtrate and a first filter residue;

[0017] (3) Then, an ethanol solution with a volume concentration of 65-80% is added to the first filter residue, and the amount of the ethanol solution added is 10-15 times the mass of the first filter residue. The mixture is heated at 30-45° C. for 2-3 h, filtered and centrifuged, and the second filtrate is collected. The first filtrate and the second filtrate are combined, concentrated, and freeze-dried to obtain the chrysanthemum extract.

[0018] Preferably, the specific preparation method of the Caulis Atractylodes macrocephala extract is as follows:

[0019] (1) Wash, dry, and crush the Tibetan Atractylodes lancea to obtain Tibetan Atractylodes lancea powder;

[0020] (2) adding 10-15 times the weight of the Tibetan wood powder to water and soaking it for 2-3 hours, and then boiling it for 3-4 hours; filtering and centrifuging to collect the first filtrate and the first filter residue;

[0021] (3) Then, 10-15 times the weight of water is added to the first filter residue and soaked for 2-3 hours, followed by decoction for 2-3 hours; after filtering and centrifugation, the second filtrate is collected, the first filtrate and the second filtrate are combined, concentrated, and freeze-dried to obtain a Tibetan Akebia extract.

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

[0023] According to the preparation method of the composite enzyme preparation for degrading mycotoxins in feed described above, the preparation method is as follows: weigh the following according to the parts by weight: mycotoxin degrading enzyme, composite probiotic powder, chrysanthemum extract, Tibetan clematis extract, and montmorillonite, and stir them evenly at room temperature to obtain the product.

[0024] The composite enzyme preparation for degrading mycotoxins in feed is used in the preparation of feed additives.

[0025] Compared with the prior art, the beneficial effects of the present invention are mainly:

[0026] The present invention prepares a composite enzyme preparation for degrading mycotoxins in feed, comprising a mycotoxin degrading enzyme, a composite probiotic powder, a chrysanthemum extract, a Tibetan clematis extract, and montmorillonite. The chrysanthemum extract and the Tibetan clematis extract have a synergistic effect and can effectively inhibit mold formation, and L-glutamyl dipeptide is added to the probiotic fermentation medium to improve the degradation efficiency of mycotoxins. The test results show that adding the composite enzyme preparation for degrading mycotoxins in feed to the feed can significantly reduce the number of molds in the feed, increase daily weight gain, reduce feed-to-weight ratio, and effectively promote the growth and development ability of animals. The composite enzyme preparation for degrading mycotoxins in feed of the present invention is green and non-toxic, meets food safety requirements, and has broad application prospects. DETAILED DESCRIPTION

[0027] The technical solution of the present invention will be further described below in conjunction with specific embodiments. However, it should be understood by those skilled in the art that the following examples are only intended to illustrate the present invention and should not be construed as limiting the present invention. Specific conditions not specified in the examples are to be followed according to conventional conditions or the conditions recommended by the manufacturer. All reagents or instruments used, unless otherwise specified, are conventional products obtained from commercial channels.

[0028] The preparation method of the Chrysanthemum morifolium extract used in the embodiment of the present invention is:

[0029] (1) Wash, dry, and crush fresh chrysanthemum to obtain chrysanthemum powder;

[0030] (2) adding the chrysanthemum powder to an ethanol solution having a volume concentration of 75%, wherein the amount of the ethanol solution added is 12 times the mass of the chrysanthemum powder, and heating and extracting at 37° C. for 2 h; filtering and centrifuging to collect a first filtrate and a first filter residue;

[0031] (3) Then, a 75% volume concentration of ethanol solution was added to the first filter residue, and the amount of ethanol solution added was 12 times the mass of the first filter residue. The mixture was heated at 37°C for 2 h, filtered and centrifuged, and the second filtrate was collected. The first filtrate and the second filtrate were combined, concentrated, and freeze-dried to obtain the chrysanthemum extract.

[0032] The preparation method of the Tibetan Akebia extract used in the embodiment of the present invention is:

[0033] (1) Wash, dry, and crush the Tibetan Atractylodes lancea to obtain Tibetan Atractylodes lancea powder;

[0034] (2) adding 12 times the weight of the Tibetan wood powder to water and soaking it for 2 hours, and then boiling it for 3 hours; filtering and centrifuging to collect the first filtrate and the first filter residue;

[0035] (3) Then, 12 times the weight of water was added to the first filter residue and soaked for 2 hours, followed by decocting for 2 hours; the second filtrate was collected after filtration and centrifugation, and the first filtrate and the second filtrate were combined, concentrated, and freeze-dried to obtain a Tibetan Akebia extract.

[0036] Example 1

[0037] A composite enzyme preparation for degrading mycotoxins in feed, comprising the following components in parts by weight: 30 parts of mycotoxin-degrading enzyme, 22 parts of composite probiotic powder, 17 parts of chrysanthemum morifolium extract, 10 parts of Tibetan clematis extract, and 13 parts of montmorillonite; the mycotoxin-degrading enzyme, in parts by weight, comprises 1 part of aflatoxin-degrading enzyme, 4 parts of zearalenone-degrading enzyme, and 3 parts of vomitoxin-degrading enzyme.

[0038] The specific preparation method of the composite 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 a Bacillus subtilis bacterial solution; Lactobacillus plantarum was inoculated into 2 mL of MRS medium and cultured at 37°C for 21 h to obtain a Lactobacillus plantarum bacterial solution; the effective viable cell count of the Bacillus subtilis bacterial solution was 2×10 8 CFU / mL; the effective viable count of the Lactobacillus plantarum liquid is 3×10 8 CFU / mL.

[0040] (2) Glucose, peptone, ammonium nitrate, tyrosine, L-glutamyl dipeptide, yeast extract, magnesium sulfate, zinc sulfate, iron sulfate, manganese sulfate, potassium dihydrogen phosphate, and Tween-80 were dissolved in deionized water according to their respective concentrations, mixed evenly, adjusted to pH 6.0, and sterilized at 115°C for 20 min to obtain a fermentation medium. The fermentation medium contained 18 g / L glucose, 9 g / L peptone, 3.0 g / L ammonium nitrate, 4.3 g / L tyrosine, 0.5 g / L L-glutamyl dipeptide, 4.2 g / L yeast extract, 0.4 g / L magnesium sulfate, 0.16 g / L zinc sulfate, 0.2 g / L iron sulfate, 0.22 g / L manganese sulfate, 3.6 g / L potassium dihydrogen phosphate, and 1.2 g / L Tween-80 in terms of final concentration.

[0041] (3) inoculating the Bacillus subtilis liquid, the Lactobacillus plantarum liquid and the fermentation medium prepared in step (2) at a volume ratio of 5:6:100, culturing at 37° C. for 35 h to obtain a fermentation product; and freeze-drying the fermentation product to obtain a composite probiotic powder.

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

[0043] The mycotoxin degrading enzyme, compound probiotic powder, chrysanthemum chinense extract, Tibetan clematis extract and montmorillonite are weighed according to the parts by weight, and stirred evenly at room temperature to obtain the product.

[0044] Example 2

[0045] A composite enzyme preparation for degrading mycotoxins in feed, comprising the following components in parts by weight: 25 parts of mycotoxin-degrading enzyme, 15 parts of composite probiotic powder, 10 parts of chrysanthemum morifolium extract, 8 parts of Tibetan clematis extract, and 6 parts of montmorillonite; the mycotoxin-degrading enzyme, in parts by weight, comprises 1 part of aflatoxin-degrading enzyme, 2 parts of zearalenone-degrading enzyme, and 1 part of vomitoxin-degrading enzyme.

[0046] The specific preparation method of the composite 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 a Bacillus subtilis bacterial solution; Lactobacillus plantarum was inoculated into 2 mL of MRS medium and cultured at 37°C for 18 h to obtain a Lactobacillus plantarum bacterial solution; the effective viable cell count of the Bacillus subtilis bacterial solution was 1×10 8 CFU / mL; the effective viable count of the Lactobacillus plantarum liquid is 1×10 8 CFU / mL.

[0048] (2) Glucose, peptone, ammonium nitrate, tyrosine, L-glutamyl dipeptide, yeast extract, magnesium sulfate, zinc sulfate, iron sulfate, manganese sulfate, potassium dihydrogen phosphate, and Tween-80 were fully dissolved in deionized water according to their respective concentrations, mixed evenly, adjusted to pH 6.0, and sterilized at 115°C for 20 minutes to obtain a fermentation medium. The fermentation medium contained 16 g / L glucose, 8 g / L peptone, 2.0 g / L ammonium nitrate, 3.0 g / L tyrosine, 0.2 g / L L-glutamyl dipeptide, 3.5 g / L yeast extract, 0.3 g / L magnesium sulfate, 0.1 g / L zinc sulfate, 0.1 g / L iron sulfate, 0.2 g / L manganese sulfate, 3.0 g / L potassium dihydrogen phosphate, and 0.5 g / L Tween-80 in terms of final concentration.

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

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

[0051] The mycotoxin degrading enzyme, compound probiotic powder, chrysanthemum chinense extract, Tibetan clematis extract and montmorillonite are weighed according to the parts by weight, and stirred evenly at room temperature to obtain the product.

[0052] Example 3

[0053] A composite enzyme preparation for degrading mycotoxins in feed, comprising the following components in parts by weight: 35 parts of mycotoxin-degrading enzyme, 30 parts of composite probiotic powder, 25 parts of chrysanthemum morifolium extract, 12 parts of Tibetan clematis extract, and 20 parts of montmorillonite; the mycotoxin-degrading enzyme, in parts by weight, comprises 2 parts of aflatoxin-degrading enzyme, 6 parts of zearalenone-degrading enzyme, and 5 parts of vomitoxin-degrading enzyme.

[0054] The specific preparation method of the composite 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 a Bacillus subtilis bacterial solution; Lactobacillus plantarum was inoculated into 2 mL of MRS medium and cultured at 37°C for 24 h to obtain a Lactobacillus plantarum bacterial solution; the effective viable cell count of the Bacillus subtilis bacterial solution was 3×10 8 CFU / mL; the effective viable count of the Lactobacillus plantarum liquid is 5×10 8 CFU / mL.

[0056] (2) Glucose, peptone, ammonium nitrate, tyrosine, L-glutamyl dipeptide, yeast extract, magnesium sulfate, zinc sulfate, iron sulfate, manganese sulfate, potassium dihydrogen phosphate, and Tween-80 were dissolved in deionized water according to their respective concentrations and mixed evenly. The pH was adjusted to 6.0, and the culture medium was sterilized at 115°C for 20 min to obtain a fermentation medium. The fermentation medium contained 21 g / L glucose, 11 g / L peptone, 4.0 g / L ammonium nitrate, 5.6 g / L tyrosine, 0.9 g / L L-glutamyl dipeptide, 5.0 g / L yeast extract, 0.6 g / L magnesium sulfate, 0.23 g / L zinc sulfate, 0.3 g / L iron sulfate, 0.25 g / L manganese sulfate, 4.2 g / L potassium dihydrogen phosphate, and 2.0 g / L Tween-80 in terms of final concentration.

[0057] (3) inoculating the Bacillus subtilis liquid, the Lactobacillus plantarum liquid and the fermentation medium prepared in step (2) at a volume ratio of 5:7:100, culturing at 37° C. for 40 h to obtain a fermentation product; and freeze-drying the fermentation product to obtain a composite probiotic powder.

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

[0059] The mycotoxin degrading enzyme, compound probiotic powder, chrysanthemum chinense extract, Tibetan clematis extract and montmorillonite are weighed according to the parts by weight, and stirred evenly at room temperature to obtain the product.

[0060] Comparative Example 1

[0061] The difference between Comparative Example 1 and Example 1 is that the chrysanthemum extract is omitted from the components of the complex enzyme preparation for degrading mycotoxins in feed, and the rest are the same as in Example 1.

[0062] Comparative Example 2

[0063] The difference between Comparative Example 2 and Example 1 is that the Tibetan Atractylodes Caulis extract is omitted from the components of the complex enzyme preparation for degrading mycotoxins in feed, and the rest are the same as Example 1.

[0064] Comparative Example 3

[0065] The difference between Comparative Example 3 and Example 1 is that the extract of Chrysanthemum morifolium in the compound enzyme preparation for degrading mycotoxins in feed is replaced with the extract of Caulis Tiliae; the other contents are the same as those in Example 1.

[0066] Comparative Example 4

[0067] The difference between Comparative Example 4 and Example 1 is that the Tibetan Akebia extract in the composite enzyme preparation for degrading mycotoxins in feed is replaced with the Chrysanthemum villosum extract, and the rest is the same as Example 1.

[0068] Comparative Example 5

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

[0070] Test Example 1

[0071] Effects of mycotoxin-degrading complex enzyme preparations on mold counts:

[0072] 9000g of fattening pig feed was weighed and divided 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 1000g per group. 10g of the complex enzyme preparation prepared in the corresponding groups of Examples 1-3 and Comparative Examples 1-5 was added to each group. No product was added to the control group. After mixing, the mixture was bagged and placed in a test chamber at a temperature of 30°C and a relative humidity of 75%. Starting from the 20th day, the growth of mold was observed and recorded daily. Samples were taken after storage for 50 days, and the total mold count in each group of feed was determined according to "GB / T13092-2006 Determination of the total mold count in feed". The total mold count results for each group of feed were recorded, as shown in Table 1.

[0073] Table 1 The number of molds in each group of feed after adding compound enzyme preparations and storing for 50 days

[0074]

[0075]

[0076] The results are shown in Table 1, which shows the number of molds in each group of feed after adding the complex enzyme preparation and storing for 50 days. As can be seen from the table, by adding the complex enzyme preparation to the feed, the complex enzyme preparations prepared in Examples 1-3 of the present invention can significantly reduce the number of molds in the feed compared with Comparative Examples 1-5. This shows that the combination of mycotoxin degrading enzyme, composite probiotic powder, chrysanthemum extract, Tibetan Atractylodes extract, and montmorillonite in the complex enzyme preparation prepared by the present invention can significantly reduce the number of molds in the feed.

[0077] Test Example 2

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

[0079] In this experiment, 90 fattening pigs weighing about 50 kg were selected and randomly divided 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, the experimental period began. Example 1 group: The composite enzyme preparation prepared in Example 1 was added to the feed at a rate of 1% by weight of the feed. The difference between the control group and Example 1 group is that the composite enzyme preparation prepared in Example 1 was not added to the feed during the feeding process, and other contents such as feeding feed were the same as those of Example 1 group; the remaining groups added the composite enzyme preparation of the corresponding group at a rate of 1% by weight of the feed to the fattening pig feed, and each group was fed continuously for 60 days. The fattening pig feed formula, in terms of mass percentage, includes the following raw materials: 65% corn, 13% wheat bran, 15% soybean meal, 1% calcium hydrogen phosphate, 2% calcium carbonate, 2% salt, and 2% compound premix for fattening pigs. All groups were treated with the same feed weight, housing conditions, and management methods. The animals were fed four times daily with ad libitum access to feed and water. The initial and final weights, daily weight gain, and feed consumption were calculated. The average initial weight, average final weight, average daily weight gain, and feed-to-weight ratio for each group were also calculated. The results are shown in Table 2.

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

[0081]

[0082]

[0083] The results are shown in Table 2, which represents an important indicator of the fattening effect of each group of pigs. As can be seen from the table, by adding the complex enzyme preparation to the feed, the complex enzyme preparations prepared in Examples 1-3 of the present invention significantly increased the daily weight gain of pigs, reduced the feed-to-weight ratio, and effectively promoted the growth and development of pigs, compared with Comparative Examples 1-5. This demonstrates that the mycotoxin-degrading complex enzyme preparations prepared in Examples 1-3 of the present invention can effectively prevent the formation of mycotoxins in feed, reduce the impact of mycotoxins on pigs, save feed costs, and improve economic benefits.

[0084] Further analysis shows that compared with the group in Example 1, the composite enzyme preparation prepared in Comparative Example 1 omitted the chrysanthemum extract; the composite enzyme preparation prepared in Comparative Example 2 omitted the Tibetan Mutong extract; the composite enzyme preparation prepared in Comparative Example 3 omitted the chrysanthemum extract, and the amount of the Tibetan Mutong extract was adjusted to the sum of the two; the composite enzyme preparation prepared in Comparative Example 4 omitted the Tibetan Mutong extract, and the amount of the chrysanthemum extract was adjusted to the sum of the two; L-glutamyl dipeptide was omitted from the fermentation medium component of Comparative Example 5; the effects of the composite enzyme preparations of the five groups on degrading mycotoxins in feed were significantly reduced. This shows that the chrysanthemum extract and the Tibetan Mutong extract in the composite enzyme preparation components have a synergistic effect; and by adding L-glutamyl dipeptide to the fermentation medium, the degradation efficiency of mycotoxins is improved; only by using them together can the stability of the composite enzyme preparation be effectively improved, and the composite enzyme preparation maintains good activity, thereby effectively preventing the formation of mycotoxins in feed.

[0085] Finally, it should be noted that the above embodiments are intended only to illustrate the technical solutions of the present invention and are not intended to limit the same. The basic principles and main features of the present invention have been described above using specific implementation schemes. Modifications or substitutions may be made based on the present invention, but such modifications or substitutions do not detract from the essence of the corresponding technical solutions from the scope of protection claimed by the present invention.

Claims

1. A composite enzyme preparation for degrading mycotoxins in feed, characterized in that: The complex enzyme preparation comprises the following components in parts by weight: 25-35 parts of mycotoxin degrading enzyme, 15-30 parts of complex probiotic powder, 10-25 parts of chrysanthemum chinense extract, 8-12 parts of Tibetan Atractylodes macrocephala extract, and 6-20 parts of montmorillonite.

2. The composite enzyme preparation for degrading mycotoxins in feed according to claim 1, characterized in that: The specific preparation method of the composite probiotic powder is as follows: after activating Bacillus subtilis and Lactobacillus plantarum respectively, Bacillus subtilis liquid and Lactobacillus plantarum liquid are obtained respectively; the Bacillus subtilis liquid and Lactobacillus plantarum liquid are inoculated into a fermentation medium and cultured at 35-40° C. for 30-40 hours to obtain a fermentation product; and the fermentation product is freeze-dried to obtain the composite probiotic powder.

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

4. The complex enzyme preparation for degrading mycotoxins in feed according to claim 2, characterized in that: The specific steps of activating Bacillus subtilis are as follows: inoculating Bacillus subtilis into LB culture medium, culturing at 35-40° C. for 24-28 hours, and obtaining Bacillus subtilis bacterial liquid; the specific steps of activating Lactobacillus plantarum are as follows: inoculating Lactobacillus plantarum into MRS culture medium, culturing at 35-40° C. for 18-24 hours, and obtaining Lactobacillus plantarum bacterial liquid.

5. The complex enzyme preparation for degrading mycotoxins in feed according to claim 2, characterized in that: The effective viable bacteria count of the Bacillus subtilis liquid is (1-3)×10 8 CFU / mL; the effective viable bacteria count of the Lactobacillus plantarum liquid is (1-5)×10 8 CFU / mL; the Bacillus subtilis bacterial solution, Lactobacillus plantarum bacterial solution and 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 in that: The specific preparation method of the chrysanthemum extract is as follows: (1) Wash, dry, and crush fresh chrysanthemum to obtain chrysanthemum powder; (2) adding the chrysanthemum powder to an ethanol solution having a volume concentration of 65-80%, wherein the amount of the ethanol solution added is 10-15 times the mass of the chrysanthemum powder, and heating and extracting at 30-45° C. for 2-3 hours; filtering and centrifuging to collect a first filtrate and a first filter residue; (3) Then, an ethanol solution with a volume concentration of 65-80% is added to the first filter residue, and the amount of the ethanol solution added is 10-15 times the mass of the first filter residue. The mixture is heated at 30-45° C. for 2-3 h, filtered and centrifuged, and the second filtrate is collected. The first filtrate and the second filtrate are combined, concentrated, and freeze-dried to obtain the chrysanthemum extract.

7. The complex enzyme preparation for degrading mycotoxins in feed according to claim 1, characterized in that: The specific preparation method of the Tibetan Atractylodes Caulis extract is as follows: (1) Wash, dry, and crush the Tibetan Atractylodes lancea to obtain Tibetan Atractylodes lancea powder; (2) adding 10-15 times the weight of the Tibetan wood powder to water and soaking it for 2-3 hours, and then boiling it for 3-4 hours; filtering and centrifuging to collect the first filtrate and the first filter residue; (3) Then, 10-15 times the weight of water is added to the first filter residue and soaked for 2-3 hours, followed by decoction for 2-3 hours; after filtering and centrifugation, the second filtrate is collected, the first filtrate and the second filtrate are combined, concentrated, and freeze-dried to obtain a Tibetan Akebia extract.

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

9. The method for preparing the composite enzyme preparation for degrading mycotoxins in feed according to any one of claims 1 to 8, characterized in that: The preparation method is as follows: weigh the following according to the parts by weight: mycotoxin degrading enzyme, compound probiotic powder, chrysanthemum chinense extract, Tibetan clematis extract, and montmorillonite, and stir them evenly at room temperature to obtain the product.

10. Use of the complex enzyme preparation for degrading mycotoxins in feed according to any one of claims 1 to 8 in the preparation of feed additives.

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