Green compound microorganism feed and preparation method thereof

Abstract

The invention relates to the technical field of microbial feeds, in particular to a green compound microbial feed and a preparation method thereof. The green compound microorganism feed comprises the following raw materials in parts by mass: 45-85 parts of pretreated soybean meal powder, 1-5 parts of zein, 0.1-0.5 part of compound trace elements and 9-24 parts of an embedding material loaded with compound enzyme, the embedding material loaded with the compound enzyme is prepared from the following raw materials in parts by mass: 1-5 parts of wheat bran, 1-3 parts of mesoporous silica, 1-2 parts of sodium butyrate, 1-2 parts of sodium bicarbonate, 1-3 parts of a compound enzyme preparation, 1-3 parts of hyaluronic acid, 1-2 parts of carboxyl-terminated polyamidoamine, 1-2 parts of gelatinized starch and 1-2 parts of citric acid. Agricultural and sideline products are used as raw materials, efficient resource conversion is achieved through microbial fermentation, dependence of traditional feed on grains is reduced, meanwhile, the preparation method is simple, and the green animal husbandry development direction is met.

Description

Technical Field

[0001] This invention relates to the field of microbial feed technology, and in particular to a green compound microbial feed and its preparation method. Background Technology

[0002] In recent years, my country's livestock industry has developed rapidly, leading to a surge in feed demand. However, the shortage of raw materials has become increasingly prominent, hindering the industry's steady development. While the complete ban on antibiotics in feed has improved food safety through antibiotic-free farming, it has also significantly increased farming costs. Against this backdrop, the development of green and safe new antibiotic alternatives has become an urgent need, giving rise to microbial fermented feeds.

[0003] Microbial fermented feed uses agricultural and food industry by-products as raw materials, adding nationally permitted beneficial microorganisms. Through fermentation, nutrients are broken down and transformed into feed ingredients that are more easily consumed, digested, and absorbed by livestock and poultry, and are non-toxic. As a new type of green feed without added drugs, it can effectively reduce the use of antibiotics and ensure the safety of livestock products.

[0004] Soybean meal, as the most widely used ingredient in animal feed, can easily trigger small intestinal hypersensitivity when consumed in large quantities by livestock and poultry, affecting the absorption of trace elements and protein digestion, thus seriously hindering breeding. However, the strains of bacteria in existing fermented feeds are mostly limited to traditional lactic acid bacteria and yeasts, with single functions and inconsistent quality. At the same time, fermented feeds are easily affected by environmental moisture, oxygen, and microorganisms, often resulting in problems such as clumping and mold growth.

[0005] Despite the significant advantages of fermented feed compared to traditional feed, its practical application still faces multiple bottlenecks. On the one hand, its limited functionality results in a limited growth-promoting effect, making it difficult to meet the diverse nutritional needs of livestock and poultry. On the other hand, its susceptibility to mold not only affects feed stability but may also pose food safety risks. These issues collectively restrict the widespread adoption of fermented feed, hindering its extensive use in the livestock industry. Summary of the Invention

[0006] The purpose of this invention is to address the shortcomings of existing technologies by proposing a green compound microbial feed and its preparation method.

[0007] A green compound microbial feed, the raw materials of which, by weight, include: 45-85 parts pretreated soybean meal powder, 1-5 parts zein, 0.1-0.5 parts compound trace elements, and 9-24 parts encapsulating material loaded with compound enzymes; the raw materials for preparing the encapsulating material loaded with compound enzymes, by weight, include: 1-5 parts wheat bran, 1-3 parts mesoporous silica, 1-2 parts sodium butyrate, 1-2 parts sodium bicarbonate, 1-3 parts compound enzyme preparation, 1-3 parts hyaluronic acid, 1-2 parts carboxyl-terminated polyamide amine, 1-2 parts gelatinized starch, and 1-2 parts citric acid.

[0008] Preferably, the encapsulating material loaded with the complex enzyme comprises: a wheat bran-mesoporous silica complex loaded with the complex enzyme, and a coating covering the outside of the wheat bran-mesoporous silica complex loaded with the complex enzyme.

[0009] More preferably, the raw materials for preparing the wheat bran-mesoporous silica composite are wheat bran, mesoporous silica, sodium butyrate, and sodium bicarbonate.

[0010] More preferably, the raw materials for preparing the coating are hyaluronic acid, carboxyl-terminated polyamide amine, gelatinized starch, and citric acid.

[0011] Preferably, the compound enzyme preparation includes: cellulase, xylanase, acidic protease, amylase, and phytase.

[0012] Preferably, the enzyme activity of cellulase is 4000-8000 U / g, the enzyme activity of xylanase is 2000-4000 U / g, the enzyme activity of acidic protease is 5000-7000 U / g, the enzyme activity of amylase is 3000-6000 U / g, and the enzyme activity of phytase is 7000-9000 U / g.

[0013] Preferably, the pretreated soybean meal powder is prepared by the following steps: crush the soybean meal, sterilize it with high pressure steam for 15-30 minutes, add water and glucose and stir evenly, inoculate with compound microbial agent for anaerobic fermentation for 2-4 days, then ferment it aerobically for 1-2 days, sterilize it at high temperature, add stone powder and stir evenly, freeze dry, and crush.

[0014] More preferably, the compound microbial agent includes: Lactobacillus plantarum, Clostridium butyricum, Bacillus licheniformis, yeast, and Bacillus subtilis.

[0015] Specifically, the viable count of *Lactobacillus plantarum* is 4-8 × 10⁻⁸. 8 The cfu / g count of Clostridium butyricum was 7-9 × 10⁻⁹. 7 CFU / g, viable count of Bacillus licheniformis is 1-3 × 10⁻⁶. 8 CFU / g, viable yeast count 8-12 × 10⁻⁶ 7 CFU / g, viable count of Bacillus subtilis is 1-3 × 10⁻⁶. 8 cfu / g.

[0016] More preferably, the mass ratio of soybean meal, glucose, and limestone powder is 50-100:1-5:1-5.

[0017] More preferably, the fermentation temperature is maintained at 25-30℃ during anaerobic fermentation and at 32-35℃ during aerobic fermentation.

[0018] The preparation method of the above-mentioned green compound microbial feed includes the following steps: S1. Add hyaluronic acid to an ethanol aqueous solution and stir until homogeneous. Add carboxyl-terminated polyamide amine and gelatinized starch. Reflux and stir at 70-90℃ for 10-30 minutes. Cool to 30-40℃ and add citric acid. Stir until homogeneous to obtain the coating material. S2. Crush wheat bran, add it to water, then add mesoporous silica, sodium butyrate, sodium bicarbonate, and compound enzyme preparation, and stir for 1-2 hours. During the stirring process, assist with ultrasonic treatment, centrifuge, and granulate. Spray coating material onto the surface of the granules, controlling the temperature at 30-40℃ during the spraying process. After uniform coating, raise the temperature to 50-60℃ and let stand for 1-2 hours to dry. Add pretreated soybean meal powder, corn gluconate, and compound trace elements, and stir evenly.

[0019] Beneficial effects: This invention utilizes a compound microbial agent to decompose large molecular proteins and polysaccharides in soybean meal into small peptides, amino acids, and short-chain fatty acids through fermentation, significantly improving the digestibility and absorption rate of nutrients by livestock and poultry. During the feeding process, it is combined with a compound enzyme preparation to further degrade anti-nutritional factors in soybean meal, release bound mineral elements, avoid small intestinal hypersensitivity reactions caused by excessive soybean meal intake, and ensure the absorption of trace elements and the efficiency of protein metabolism.

[0020] The coating material used in this invention utilizes hyaluronic acid, gelatinized starch, and carboxyl-terminated polyamide amine to form a three-dimensional cross-linked network. The compound enzyme preparation and sodium butyrate work synergistically, and after loading, they are coated with the coating material to form a protective layer for the sodium butyrate and compound enzyme preparation, ensuring the stability of the enzyme preparation and its survival rate in the digestive tract. Sodium bicarbonate, in combination with citric acid in the coating material, promotes the formation of a microporous structure in the protective layer. Combined with the response of gelatinized starch and carboxyl-terminated polyamide amine in an acidic environment, this not only provides a gradual and sustained release effect on the internal substances but also reduces the erosion of feed by moisture or oxygen, extending the shelf life.

[0021] This invention utilizes agricultural by-products as raw materials and achieves efficient resource conversion through microbial fermentation, reducing the dependence of traditional feed on grains. At the same time, the preparation method is simple and in line with the development direction of green animal husbandry. Attached Figure Description

[0022] Figure 1 The cumulative release rate curves of sodium butyrate in artificial gastric fluid are shown for the compound microbial feeds obtained in Example 5 and Comparative Examples 1-3.

[0023] Figure 2 This is a comparison chart of the average egg weight, egg production rate, and qualified egg rate of Example 5 group and Comparative Examples 1-3 groups.

[0024] Figure 3 This is a comparison chart of eggshell thickness, yolk specific gravity, and eggshell specific gravity for Example 5 and Comparative Examples 1-3. Detailed Implementation

[0025] The present invention will be further explained below with reference to specific embodiments.

[0026] The following trace elements were purchased from Shandong Mouju Biotechnology Co., Ltd., specifically a mixture of amino acid-chelated calcium, magnesium, zinc, and selenium. The following mesoporous silica was purchased from Hangzhou Mouyou New Materials Co., Ltd., with a particle size of 6-10 μm, a pore size of 50 nm, and a whiteness of 92%. The following carboxyl-terminated polyamide amine (PAMAM-COOH, G1.0) was purchased from Weihai Mouyuan Molecular New Materials Co., Ltd., with ethylenediamine as the core. The following complex enzymes were purchased from Mousheng (Beijing) Biotechnology Development Co., Ltd., and the complex enzyme preparation included: cellulase with an activity of 6000 U / g, xylanase with an activity of 3000 U / g, acidic protease with an activity of 6000 U / g, amylase with an activity of 4500 U / g, and phytase with an activity of 8000 U / g.

[0027] Example 1

[0028] A green compound microbial feed, the raw materials of which include: 4500g of pretreated soybean meal powder, 100g of corn gliadin, 10g of compound trace elements, and 900g of encapsulated material loaded with compound enzymes.

[0029] The pretreated soybean meal powder is prepared by the following steps: 5000g of soybean meal is crushed and passed through a 60-mesh sieve, sterilized by high-pressure steam for 15 minutes, 4000g of water and 100g of glucose are added and stirred evenly, and then inoculated with compound microbial agent for anaerobic fermentation for 2 days, maintaining the fermentation temperature at 25℃ during the anaerobic fermentation process; then aerobic fermentation is carried out for 1 day, maintaining the fermentation temperature at 32℃ during the aerobic fermentation process; after high-temperature sterilization, 100g of limestone powder is added and stirred evenly, freeze-dried, and then crushed.

[0030] The viable count of *Lactobacillus plantarum* in the compound microbial agent is 4 × 10⁻⁶. 8 cfu / g, viable count of Clostridium butyricum was 7 × 10⁻⁶. 7 CFU / g, viable count of Bacillus licheniformis is 1×10⁻⁶. 8 cfu / g, viable yeast count 8×10 7 CFU / g, viable count of Bacillus subtilis is 1×10⁻⁶. 8 cfu / g.

[0031] The encapsulating material loaded with the complex enzyme comprises: a wheat bran-mesoporous silica complex loaded with the complex enzyme, and a coating covering the outer surface of the wheat bran-mesoporous silica complex loaded with the complex enzyme. The raw materials for preparing the encapsulating material loaded with the complex enzyme include: 100g wheat bran, 100g mesoporous silica, 100g sodium butyrate, 100g sodium bicarbonate, 100g complex enzyme preparation, 100g hyaluronic acid, 100g carboxyl-terminated polyamide amine, 100g gelatinized starch, and 100g citric acid.

[0032] The preparation method of the above-mentioned green compound microbial feed includes the following steps: S1. Add hyaluronic acid to 2000g of 40% ethanol aqueous solution and stir evenly. Add carboxyl-terminated polyamide amine and gelatinized starch. Reflux and stir at 70℃ for 10min. Cool down to 30℃ and add citric acid. Stir evenly to obtain the coating material. S2. Crush wheat bran through a 100-mesh sieve, add it to 2000g of water, then add mesoporous silica, sodium butyrate, sodium bicarbonate, and compound enzyme preparation. Stir for 1 hour at a stirring speed of 100r / min, and perform ultrasonic treatment during the stirring process at a frequency of 60kHz. Centrifuge and granulate. Spray coating material onto the surface of the granules, controlling the temperature at 30℃ during the spraying process. After uniform coating, raise the temperature to 50℃ and let stand for 1 hour to dry. Add pretreated soybean meal powder, corn gliadin, and compound trace elements and stir evenly.

[0033] Example 2

[0034] A green compound microbial feed, the raw materials of which include: 8500g of pretreated soybean meal powder, 500g of corn gluconate, 50g of compound trace elements, and 2400g of encapsulated material loaded with compound enzymes.

[0035] The pretreated soybean meal powder is prepared by the following steps: 10 kg of soybean meal is crushed and passed through a 100-mesh sieve, sterilized by high-pressure steam for 30 min, 6000 g of water and 500 g of glucose are added and stirred evenly, and then inoculated with compound microbial agent for anaerobic fermentation for 4 days, maintaining the fermentation temperature at 30℃ during the anaerobic fermentation process; then aerobic fermentation is carried out for 2 days, maintaining the fermentation temperature at 35℃ during the aerobic fermentation process; after high-temperature sterilization, 500 g of limestone powder is added and stirred evenly, then freeze-dried and crushed.

[0036] In the compound microbial agent, the viable count of Lactobacillus plantarum was 8 × 10⁻⁶. 8 cfu / g, viable count of Clostridium butyricum was 9 × 10⁻⁶. 7 CFU / g, viable count of Bacillus licheniformis is 3 × 10⁻⁶. 8 cfu / g, viable yeast count 12×10 7 CFU / g, viable count of Bacillus subtilis is 3 × 10⁻⁶. 8 cfu / g.

[0037] The encapsulating material loaded with the complex enzyme comprises: a wheat bran-mesoporous silica complex loaded with the complex enzyme, and a coating covering the outer surface of the wheat bran-mesoporous silica complex loaded with the complex enzyme. The raw materials for preparing the encapsulating material loaded with the complex enzyme include: 500g wheat bran, 300g mesoporous silica, 200g sodium butyrate, 200g sodium bicarbonate, 300g complex enzyme preparation, 300g hyaluronic acid, 200g carboxyl-terminated polyamide amine, 200g gelatinized starch, and 200g citric acid.

[0038] The preparation method of the above-mentioned green compound microbial feed includes the following steps: S1. Hyaluronic acid is added to 4000g of 60% ethanol aqueous solution and stirred evenly. Carboxyl-terminated polyamide amine and gelatinized starch are added. The mixture is refluxed and stirred at 90℃ for 30min. The temperature is then lowered to 40℃ and citric acid is added and stirred evenly to obtain the coating material. S2. Crush wheat bran through a 100-mesh sieve, add it to 4000g of water, then add mesoporous silica, sodium butyrate, sodium bicarbonate, and compound enzyme preparation. Stir for 2 hours at a stirring speed of 500r / min, and perform ultrasonic treatment during the stirring process at a frequency of 80kHz. Centrifuge and granulate. Spray coating material onto the surface of the granules, controlling the temperature at 40℃ during the spraying process. After uniform coating, raise the temperature to 60℃ and let stand for 2 hours to dry. Add pretreated soybean meal powder, corn gliadin, and compound trace elements and stir evenly.

[0039] Example 3

[0040] A green compound microbial feed, the raw materials of which include: 5500g of pretreated soybean meal powder, 400g of corn gliadin, 20g of compound trace elements, and 2000g of encapsulated material loaded with compound enzymes.

[0041] The pretreated soybean meal powder is prepared using the following steps: 7000g of soybean meal is pulverized through an 80-mesh sieve, sterilized by high-pressure steam for 15 minutes, 5500g of water and 200g of glucose are added and stirred evenly, inoculated with compound microbial agent and anaerobic fermented for 3.5 days, maintaining the fermentation temperature at 26℃ during the anaerobic fermentation process; then aerobic fermentation is carried out for 1.5 days, maintaining the fermentation temperature at 34℃ during the aerobic fermentation process; high-temperature sterilization is performed, 200g of limestone powder is added and stirred evenly, freeze-dried, and pulverized.

[0042] The viable count of *Lactobacillus plantarum* in the compound microbial agent is 7 × 10⁻⁶. 8 The cfu / g count of Clostridium butyricum was 7.5 × 10⁻⁶. 7 The cfu / g count of Bacillus licheniformis was 2.5 × 10⁻⁶. 8 CFU / g, viable yeast count 9 × 10⁻⁶ 7 The cfu / g count of Bacillus subtilis was 2.5 × 10⁻⁶. 8cfu / g.

[0043] The encapsulating material loaded with the complex enzyme comprises: a wheat bran-mesoporous silica complex loaded with the complex enzyme, and a coating covering the outer surface of the wheat bran-mesoporous silica complex loaded with the complex enzyme. The raw materials for preparing the encapsulating material loaded with the complex enzyme include: 400g wheat bran, 150g mesoporous silica, 170g sodium butyrate, 130g sodium bicarbonate, 250g complex enzyme preparation, 150g hyaluronic acid, 180g carboxyl-terminated polyamide amine, 120g gelatinized starch, and 150g citric acid.

[0044] The preparation method of the above-mentioned green compound microbial feed includes the following steps: S1. Hyaluronic acid is added to 3500g of 45% ethanol aqueous solution and stirred evenly. Carboxyl-terminated polyamide amine and gelatinized starch are added. The mixture is refluxed and stirred at 85℃ for 15min. The temperature is then lowered to 37℃ and citric acid is added and stirred evenly to obtain the coating material. S2. Crush wheat bran through a 100-mesh sieve, add it to 2500g of water, then add mesoporous silica, sodium butyrate, sodium bicarbonate, and compound enzyme preparation. Stir for 100min at a stirring speed of 200r / min, and perform ultrasonic treatment during stirring at a frequency of 75kHz. Centrifuge and granulate. Spray coating material onto the surface of the granules, controlling the temperature at 33℃ during spraying. After uniform coating, raise the temperature to 58℃ and let stand for 80min to dry. Add pretreated soybean meal powder, corn gliadin, and compound trace elements and stir evenly.

[0045] Example 4

[0046] A green compound microbial feed, the raw materials of which include: 7500g of pretreated soybean meal powder, 200g of corn gluconate, 40g of compound trace elements, and 1200g of encapsulated material loaded with compound enzymes.

[0047] The pretreated soybean meal powder is prepared using the following steps: 9000g of soybean meal is pulverized through an 80-mesh sieve, sterilized by high-pressure steam for 25 minutes, 4500g of water and 400g of glucose are added and stirred evenly, inoculated with compound microbial agent and anaerobic fermented for 2.5 days, maintaining the fermentation temperature at 28℃ during the anaerobic fermentation process; then aerobic fermentation is carried out for 1.5 days, maintaining the fermentation temperature at 33℃ during the aerobic fermentation process; high-temperature sterilization is performed, 400g of limestone powder is added and stirred evenly, freeze-dried, and pulverized.

[0048] In the compound microbial agent, the viable count of Lactobacillus plantarum is 5 × 10⁻⁶. 8 The cfu / g count of Clostridium butyricum was 8.5 × 10⁻⁶. 7 The cfu / g count of Bacillus licheniformis was 1.5 × 10⁻⁶. 8 cfu / g, viable yeast count was 11 × 10⁻⁶. 7The cfu / g count of Bacillus subtilis was 1.5 × 10⁻⁶. 8 cfu / g.

[0049] The encapsulating material loaded with the complex enzyme comprises: a wheat bran-mesoporous silica complex loaded with the complex enzyme, and a coating covering the outer surface of the wheat bran-mesoporous silica complex loaded with the complex enzyme. The raw materials for preparing the encapsulating material loaded with the complex enzyme include: 200g wheat bran, 250g mesoporous silica, 130g sodium butyrate, 170g sodium bicarbonate, 150g complex enzyme preparation, 250g hyaluronic acid, 120g carboxyl-terminated polyamide amine, 180g gelatinized starch, and 150g citric acid.

[0050] The preparation method of the above-mentioned green compound microbial feed includes the following steps: S1. Hyaluronic acid is added to 2500g of 55% ethanol aqueous solution and stirred evenly. Carboxyl-terminated polyamide amine and gelatinized starch are added. The mixture is refluxed and stirred at 75℃ for 25min. The temperature is then lowered to 33℃ and citric acid is added and stirred evenly to obtain the coating material. S2. Crush wheat bran through a 100-mesh sieve, add it to 3500g of water, then add mesoporous silica, sodium butyrate, sodium bicarbonate, and compound enzyme preparation. Stir for 80 minutes at a stirring speed of 400r / min, and perform ultrasonic treatment during stirring at a frequency of 65kHz. Centrifuge and granulate. Spray coating material onto the surface of the granules, controlling the temperature at 37℃ during spraying. After uniform coating, raise the temperature to 52℃ and let stand for 100 minutes to dry. Add pretreated soybean meal powder, corn gluconate, and compound trace elements and stir evenly.

[0051] Example 5

[0052] A green compound microbial feed, the raw materials of which include: 6500g of pretreated soybean meal powder, 300g of corn gliadin, 30g of compound trace elements, and 1600g of encapsulated material loaded with compound enzymes.

[0053] The pretreated soybean meal powder is prepared by the following steps: 8000g of soybean meal is crushed and passed through an 80-mesh sieve, sterilized by high-pressure steam for 20 minutes, 5000g of water and 300g of glucose are added and stirred evenly, inoculated with compound microbial agent for anaerobic fermentation for 3 days, and the fermentation temperature is maintained at 27℃ during the anaerobic fermentation process; then aerobic fermentation is carried out for 1.5 days, and the fermentation temperature is maintained at 33℃ during the aerobic fermentation process; high-temperature sterilization is performed, 300g of limestone powder is added and stirred evenly, freeze-dried, and crushed.

[0054] In the compound microbial agent, the viable count of Lactobacillus plantarum is 6 × 10⁻⁶. 8 cfu / g, viable count of Clostridium butyricum was 8 × 10⁻⁶. 7 CFU / g, viable count of Bacillus licheniformis is 2 × 10⁻⁶. 8cfu / g, viable yeast count 1×10 8 CFU / g, viable count of Bacillus subtilis is 2 × 10⁻⁶. 8 cfu / g.

[0055] The encapsulating material loaded with the complex enzyme comprises: a wheat bran-mesoporous silica complex loaded with the complex enzyme, and a coating covering the outer surface of the wheat bran-mesoporous silica complex loaded with the complex enzyme. The raw materials for preparing the encapsulating material loaded with the complex enzyme include: 300g wheat bran, 200g mesoporous silica, 150g sodium butyrate, 150g sodium bicarbonate, 200g complex enzyme preparation, 200g hyaluronic acid, 150g carboxyl-terminated polyamide amine, 150g gelatinized starch, and 150g citric acid.

[0056] The preparation method of the above-mentioned green compound microbial feed includes the following steps: S1. Add hyaluronic acid to 3000g of 50% ethanol aqueous solution and stir evenly. Add carboxyl-terminated polyamide amine and gelatinized starch. Reflux and stir at 80℃ for 20min. Cool down to 35℃ and add citric acid. Stir evenly to obtain the coating material. S2. Crush wheat bran through a 100-mesh sieve, add it to 3000g of water, then add mesoporous silica, sodium butyrate, sodium bicarbonate, and compound enzyme preparation. Stir for 90 minutes at a stirring speed of 300r / min, and perform ultrasonic treatment during stirring at a frequency of 70kHz. Centrifuge and granulate. Spray coating material onto the surface of the granules, controlling the temperature at 35℃ during spraying. After uniform coating, raise the temperature to 55℃ and let stand for 90 minutes to dry. Add pretreated soybean meal powder, corn gliadin, and compound trace elements and stir evenly.

[0057] Comparative Example 1

[0058] A green compound microbial feed, the raw materials of which include: 6500g of pretreated soybean meal powder, 300g of corn gliadin, 30g of compound trace elements, and 1600g of encapsulated material loaded with compound enzymes.

[0059] The pretreated soybean meal powder is prepared by the following steps: 8000g of soybean meal is crushed and passed through an 80-mesh sieve, sterilized by high-pressure steam for 20 minutes, 5000g of water and 300g of glucose are added and stirred evenly, inoculated with compound microbial agent for anaerobic fermentation for 3 days, and the fermentation temperature is maintained at 27℃ during the anaerobic fermentation process; then aerobic fermentation is carried out for 1.5 days, and the fermentation temperature is maintained at 33℃ during the aerobic fermentation process; high-temperature sterilization is performed, 300g of limestone powder is added and stirred evenly, freeze-dried, and crushed.

[0060] In the compound microbial agent, the viable count of Lactobacillus plantarum is 6 × 10⁻⁶. 8 cfu / g, viable count of Clostridium butyricum was 8 × 10⁻⁶. 7CFU / g, viable count of Bacillus licheniformis is 2 × 10⁻⁶. 8 cfu / g, viable yeast count 1×10 8 CFU / g, viable count of Bacillus subtilis is 2 × 10⁻⁶. 8 cfu / g.

[0061] The encapsulating material loaded with the complex enzyme comprises: a wheat bran-mesoporous silica complex loaded with the complex enzyme, and a coating covering the outer surface of the wheat bran-mesoporous silica complex loaded with the complex enzyme. The raw materials for preparing the encapsulating material loaded with the complex enzyme include: 300g wheat bran, 300g mesoporous silica, 150g sodium butyrate, 200g complex enzyme preparation, 200g hyaluronic acid, 150g carboxyl-terminated polyamide amine, 150g gelatinized starch, and 200g citric acid.

[0062] The preparation method of the above-mentioned green compound microbial feed includes the following steps: S1. Add hyaluronic acid to 3000g of 50% ethanol aqueous solution and stir evenly. Add carboxyl-terminated polyamide amine and gelatinized starch. Reflux and stir at 80℃ for 20min. Cool down to 35℃ and add citric acid. Stir evenly to obtain the coating material. S2. Crush wheat bran through a 100-mesh sieve, add it to 3000g of water, then add mesoporous silica, sodium butyrate, and compound enzyme preparation. Stir for 90 minutes at a stirring speed of 300r / min, and perform ultrasonic treatment during the stirring process at a frequency of 70kHz. Centrifuge and granulate. Spray coating material onto the surface of the granules, controlling the temperature at 35℃ during the spraying process. After uniform coating, raise the temperature to 55℃ and let stand for 90 minutes to dry. Add pretreated soybean meal powder, corn gluconate, and compound trace elements and stir evenly.

[0063] Comparative Example 2

[0064] A green compound microbial feed, the raw materials of which include: 6500g of pretreated soybean meal powder, 300g of corn gliadin, 30g of compound trace elements, and 1600g of encapsulated material loaded with compound enzymes.

[0065] The pretreated soybean meal powder is prepared by the following steps: 8000g of soybean meal is crushed and passed through an 80-mesh sieve, sterilized by high-pressure steam for 20 minutes, 5000g of water and 300g of glucose are added and stirred evenly, inoculated with compound microbial agent for anaerobic fermentation for 3 days, and the fermentation temperature is maintained at 27℃ during the anaerobic fermentation process; then aerobic fermentation is carried out for 1.5 days, and the fermentation temperature is maintained at 33℃ during the aerobic fermentation process; high-temperature sterilization is performed, 300g of limestone powder is added and stirred evenly, freeze-dried, and crushed.

[0066] In the compound microbial agent, the viable count of Lactobacillus plantarum is 6 × 10⁻⁶. 8 CFU / g, viable count of Bacillus licheniformis is 2 × 10⁻⁶.8 cfu / g, viable yeast count 1×10 8 CFU / g, viable count of Bacillus subtilis is 2 × 10⁻⁶. 8 cfu / g.

[0067] The encapsulating material loaded with the complex enzyme comprises: a wheat bran-mesoporous silica complex loaded with the complex enzyme, and a coating covering the outer surface of the wheat bran-mesoporous silica complex loaded with the complex enzyme. The raw materials for preparing the encapsulating material loaded with the complex enzyme include: 300g wheat bran, 200g mesoporous silica, 150g sodium butyrate, 150g sodium bicarbonate, 200g complex enzyme preparation, 200g hyaluronic acid, 150g carboxyl-terminated polyamide amine, 150g gelatinized starch, and 150g citric acid.

[0068] The preparation method of the above-mentioned green compound microbial feed includes the following steps: S1. Add hyaluronic acid to 3000g of 50% ethanol aqueous solution and stir evenly. Add carboxyl-terminated polyamide amine and gelatinized starch. Reflux and stir at 80℃ for 20min. Cool down to 35℃ and add citric acid. Stir evenly to obtain the coating material. S2. Crush wheat bran through a 100-mesh sieve, add it to 3000g of water, then add mesoporous silica, sodium butyrate, sodium bicarbonate, and compound enzyme preparation. Stir for 90 minutes at a stirring speed of 300r / min, and perform ultrasonic treatment during stirring at a frequency of 70kHz. Centrifuge and granulate. Spray coating material onto the surface of the granules, controlling the temperature at 35℃ during spraying. After uniform coating, raise the temperature to 55℃ and let stand for 90 minutes to dry. Add pretreated soybean meal powder, corn gliadin, and compound trace elements and stir evenly.

[0069] Comparative Example 3

[0070] A green compound microbial feed, the raw materials of which include: 6500g of pretreated soybean meal powder, 300g of corn gliadin, 30g of compound trace elements, and 1600g of encapsulated material loaded with compound enzymes.

[0071] The pretreated soybean meal powder is prepared by the following steps: 8000g of soybean meal is crushed and passed through an 80-mesh sieve, sterilized by high-pressure steam for 20 minutes, 5000g of water and 300g of glucose are added and stirred evenly, inoculated with compound microbial agent for anaerobic fermentation for 3 days, and the fermentation temperature is maintained at 27℃ during the anaerobic fermentation process; then aerobic fermentation is carried out for 1.5 days, and the fermentation temperature is maintained at 33℃ during the aerobic fermentation process; high-temperature sterilization is performed, 300g of limestone powder is added and stirred evenly, freeze-dried, and crushed.

[0072] In the compound microbial agent, the viable count of Lactobacillus plantarum is 6 × 10⁻⁶. 8 cfu / g, viable count of Clostridium butyricum was 8 × 10⁻⁶. 7CFU / g, viable count of Bacillus licheniformis is 2 × 10⁻⁶. 8 cfu / g, viable yeast count 1×10 8 CFU / g, viable count of Bacillus subtilis is 2 × 10⁻⁶. 8 cfu / g.

[0073] The encapsulating material loaded with the complex enzyme comprises: a wheat bran-mesoporous silica complex loaded with the complex enzyme, and a coating covering the outer surface of the wheat bran-mesoporous silica complex loaded with the complex enzyme. The raw materials for preparing the encapsulating material loaded with the complex enzyme include: 300g wheat bran, 200g mesoporous silica, 150g sodium butyrate, 150g sodium bicarbonate, 200g complex enzyme preparation, 300g hyaluronic acid, 200g gelatinized starch, and 150g citric acid.

[0074] The preparation method of the above-mentioned green compound microbial feed includes the following steps: S1. Add hyaluronic acid to 3000g of 50% ethanol aqueous solution and stir evenly. Add gelatinized starch and reflux at 80℃ for 20min. Cool down to 35℃ and add citric acid and stir evenly to obtain the coating material. S2. Crush wheat bran through a 100-mesh sieve, add it to 3000g of water, then add mesoporous silica, sodium butyrate, sodium bicarbonate, and compound enzyme preparation. Stir for 90 minutes at a stirring speed of 300r / min, and perform ultrasonic treatment during stirring at a frequency of 70kHz. Centrifuge and granulate. Spray coating material onto the surface of the granules, controlling the temperature at 35℃ during spraying. After uniform coating, raise the temperature to 55℃ and let stand for 90 minutes to dry. Add pretreated soybean meal powder, corn gliadin, and compound trace elements and stir evenly.

[0075] The compound microbial feeds obtained in Example 5 and Comparative Examples 1-3 were added to artificial gastric fluid (37°C), and samples were taken at different time points. After sampling, an equal amount of fresh artificial gastric fluid was quickly added. The sodium butyrate content in the samples was determined, and the cumulative release rate of sodium butyrate was calculated.

[0076] like Figure 1 As shown, the cumulative release rate curves of sodium butyrate of the compound microbial feed obtained in Example 5 and Comparative Example 2 are similar, and are significantly lower than those of Comparative Example and Comparative Example 3.

[0077] The compound microbial feed obtained in Example 5 and Comparative Examples 1-3 was used to conduct a feeding experiment on laying hens. The specific details are as follows: This experiment was conducted in a large chicken farm in Huanggang. Healthy Hy-Line Brown laying hens aged 24 weeks were selected for the experiment. Natural ventilation and artificial lighting (16L:8D) were used. During the experiment, the temperature in the chicken house was 28±2℃ and the relative humidity was 55-62%. The experiment was divided into 4 groups, with 7 replicates in each group and 12 chickens in each replicate. The pre-trial period was 7 days and the formal trial period was 54 days.

[0078] 1. Production indicator measurement

[0079] During the experiment, the total number of eggs laid, total egg weight, feed intake, and number of qualified eggs were recorded daily on a repeat basis. The average egg weight, egg production rate, and qualified egg rate of each group were calculated.

[0080] Average egg weight = total egg weight (g) / number of eggs laid.

[0081] Egg production rate = number of eggs produced / number of chickens × 100%.

[0082] Qualified egg rate = (Number of qualified eggs / Total number of eggs) × 100%.

[0083] like Figure 2 As shown, the average egg weight, egg production rate, and qualified egg rate of Example 5 group were the highest. The average egg weight and qualified egg rate were significantly better than those of Comparative Examples 1-3, while the egg production rate did not show a significant difference from that of Comparative Examples 1-3.

[0084] 2. Egg quality determination

[0085] Egg samples were collected one day before the end of the experiment. Four eggs with good appearance and similar weight were randomly selected for each replicate. The weight and shell thickness (average of blunt end, pointed end and middle) of each egg were weighed. The yolk and egg white were separated, and the weight of the yolk, egg white and egg shell were weighed and their respective proportions were calculated.

[0086] like Figure 3 As shown, the eggshell thickness, yolk specific gravity, and eggshell specific gravity of the 5th example group were the highest, significantly better than those of the 1st to 3rd comparative examples.

[0087] The reason for the above results is that this invention utilizes a compound microbial agent to decompose the large molecular proteins and polysaccharides in soybean meal into small peptides, amino acids, and short-chain fatty acids through fermentation, significantly improving the digestibility and absorption rate of nutrients by livestock and poultry. During the feeding process, it is combined with a compound enzyme preparation to further degrade anti-nutritional factors in soybean meal, releasing bound mineral elements and avoiding small intestinal hypersensitivity reactions caused by excessive soybean meal intake, thus ensuring the absorption of trace elements and the efficiency of protein metabolism. The coating material used in this invention utilizes hyaluronic acid, gelatinized starch, and carboxyl-terminated polyamide amine to form a three-dimensional cross-linked network. The compound enzyme preparation and sodium butyrate work synergistically, and after loading, are coated with the coating material to form a protective layer for the sodium butyrate and compound enzyme preparation, ensuring the stability of the enzyme preparation and its survival rate in the digestive tract. Sodium bicarbonate, in combination with citric acid in the coating material, promotes the formation of a microporous structure in the protective layer. Combined with the response of gelatinized starch and carboxyl-terminated polyamide amine in an acidic environment, this not only provides a gradual and sustained release effect on the internal substances but also reduces the erosion of feed by moisture or oxygen, extending shelf life.

[0088] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. A green compound microbial feed, characterized in that, The raw materials, by weight, include: 45-85 parts pretreated soybean meal powder, 1-5 parts zein, 0.1-0.5 parts compound trace elements, and 9-24 parts encapsulating material loaded with compound enzymes. The raw materials for preparing the encapsulating material loaded with complex enzymes include, by weight, 1-5 parts wheat bran, 1-3 parts mesoporous silica, 1-2 parts sodium butyrate, 1-2 parts sodium bicarbonate, 1-3 parts complex enzyme preparation, 1-3 parts hyaluronic acid, 1-2 parts carboxyl-terminated polyamide amine, 1-2 parts gelatinized starch, and 1-2 parts citric acid.

2. The green compound microbial feed according to claim 1, characterized in that, The embedding material loaded with the complex enzyme includes: a wheat bran-mesoporous silica complex loaded with the complex enzyme, and a coating on the outside of the wheat bran-mesoporous silica complex loaded with the complex enzyme. The raw materials for preparing the wheat bran-mesoporous silica composite are wheat bran, mesoporous silica, sodium butyrate, and sodium bicarbonate; the raw materials for preparing the coating are hyaluronic acid, carboxyl-terminated polyamide amine, gelatinized starch, and citric acid.

3. The green compound microbial feed according to claim 1, characterized in that, The compound enzyme preparation includes: cellulase, xylanase, acidic protease, amylase, and phytase.

4. The green compound microbial feed according to claim 3, characterized in that, The enzyme activity of cellulase is 4000-8000 U / g, that of xylanase is 2000-4000 U / g, that of acidic protease is 5000-7000 U / g, that of amylase is 3000-6000 U / g, and that of phytase is 7000-9000 U / g.

5. The green compound microbial feed according to claim 1, characterized in that, The pretreated soybean meal powder is prepared by the following steps: crush the soybean meal, sterilize it with high pressure steam for 15-30 minutes, add water and glucose and stir evenly, inoculate with compound microbial agent for anaerobic fermentation for 2-4 days, then ferment aerobically for 1-2 days, sterilize at high temperature, add stone powder and stir evenly, freeze dry, and crush.

6. The green compound microbial feed according to claim 5, characterized in that, The compound microbial agent includes: Lactobacillus plantarum, Clostridium butyricum, Bacillus licheniformis, yeast, and Bacillus subtilis.

7. The green compound microbial feed according to claim 6, characterized in that, The viable count of Lactobacillus plantarum is 4-8 × 10⁻⁸. 8 The cfu / g count of Clostridium butyricum was 7-9 × 10⁻⁹. 7 CFU / g, viable count of Bacillus licheniformis is 1-3 × 10⁻⁶. 8 CFU / g, viable yeast count 8-12 × 10⁻⁶ 7 CFU / g, viable count of Bacillus subtilis is 1-3 × 10⁻⁶. 8 cfu / g.

8. The green compound microbial feed according to claim 5, characterized in that, The mass ratio of soybean meal, glucose, and limestone powder is 50-100:1-5:1-5.

9. The green compound microbial feed according to claim 5, characterized in that, During anaerobic fermentation, maintain the fermentation temperature at 25-30℃; during aerobic fermentation, maintain the fermentation temperature at 32-35℃.

10. A method for preparing a green compound microbial feed as described in any one of claims 1-9, characterized in that, Includes the following steps: S1. Add hyaluronic acid to an ethanol aqueous solution and stir until homogeneous. Add carboxyl-terminated polyamide amine and gelatinized starch. Reflux and stir at 70-90℃ for 10-30 minutes. Cool to 30-40℃ and add citric acid. Stir until homogeneous to obtain the coating material. S2. Crush wheat bran, add it to water, then add mesoporous silica, sodium butyrate, sodium bicarbonate, and compound enzyme preparation, and stir for 1-2 hours. During the stirring process, assist with ultrasonic treatment, centrifuge, and granulate. Spray coating material onto the surface of the granules, controlling the temperature at 30-40℃ during the spraying process. After uniform coating, raise the temperature to 50-60℃ and let stand for 1-2 hours to dry. Add pretreated soybean meal powder, corn gluconate, and compound trace elements, and stir evenly.

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