Fermented straw feed protein additive, preparation method and application thereof

Abstract

The application discloses a fermented straw feed protein additive and a preparation method and application thereof, and belongs to the technical field of feed additives. The application provides a preparation method of a fermented straw feed protein additive, wherein dandelion and jerusalem artichoke straws are used as basic raw materials, steam explosion and sterilization are carried out, then the steam explosion and sterilization products are mixed with organic nitrogen sources, first fermentation is carried out by using a first composite bacteria, the fermentation products are used as a culture medium for culturing composite bacteria mycelia, after the mycelia grow to be full, second fermentation is carried out by using a second composite bacteria, and the second fermentation products are dried, crushed and then the fermented straw feed protein additive is obtained. The fermented straw feed protein additive has the characteristics of less pollution, high efficiency, low cost and being beneficial to industrialized production, the protein content of the additive can be more than 20%, the crude fiber degradation rate is more than 30%, and the phenolic acid content can be more than 7%. The problems that agricultural straws are not fully utilized, the digestion and absorption rate of laying hens, immunity, antioxidant capacity and egg quality improvement are not significant in the prior art are solved.

Description

Technical Field

[0001] This invention belongs to the field of feed additive technology, specifically relating to a fermented straw feed protein additive, its preparation method, and its application. Background Technology

[0002] Studies have shown that various straws, vines, and branches and leaves of non-toxic plants contain many effective components that can be consumed by livestock. However, because these straws are high in cellulose and hemicellulose and are difficult to digest, they are discarded as waste.

[0003] As people's living standards improve, the demand for animal products is also increasing, which necessitates the rapid development of animal husbandry. Feed is the foundation of animal husbandry, and its development largely depends on the development of the feed industry. However, the supply-demand gap for conventional feed resources such as grains, oilseed cakes, and forage is widening, and excessive reliance on grains as feed cannot meet the needs of sustainable development in animal husbandry. Therefore, how to utilize straw to prepare feed or feed additives is a current technical challenge. Summary of the Invention

[0004] The purpose of this invention is to provide a fermented straw feed protein additive, its preparation method and application, which has the characteristics of low pollution, high efficiency, low cost and is conducive to industrial production, while significantly improving the nutritional content of feed.

[0005] This invention provides a method for preparing a fermented straw feed protein additive, comprising the following steps:

[0006] (1) Dandelion and Jerusalem artichoke stalks were crushed and then steam-exploded to obtain pretreated base material;

[0007] (2) After mixing the pretreated substrate described in step (1) with an organic nitrogen source, the first compound bacteria are inoculated to carry out the first fermentation to obtain the degraded material; the first compound bacteria include termite fungi, Trichoderma longipes, Phanerochaete chrysosporium, Aspergillus niger and Bacillus subtilis;

[0008] (3) After inoculating the degradation material described in step (2) with composite mycelium, culture is carried out to obtain a high mycelium protein material; the composite mycelium includes mycelium of Russula rubra and Codonopsis pilosula;

[0009] (4) The high microbial protein material described in step (3) is fermented using the second compound bacteria to obtain fermented high microbial protein material, which is then dried and pulverized to obtain the fermented straw feed protein additive; the second compound bacteria include brewer's yeast and Bifidobacterium.

[0010] Preferably, the mass ratio of dandelion and Jerusalem artichoke straw in step (1) is (0.1-1.5):(10-25).

[0011] Preferably, the mass of the organic nitrogen source in step (2) is 0.5% to 2% of the mass of the pretreated base material;

[0012] Preferably, the organic nitrogen source includes a composite organic nitrogen source, which, by mass parts, includes: 5-10 parts yeast powder, 5-10 parts corn steep liquor powder, 5-10 parts soybean meal powder, and 5-10 parts peptone.

[0013] Preferably, in step (2), the temperature of the first fermentation is 37-40°C, and the fermentation time is 5-7 days;

[0014] Preferably, the inoculum amount of the first compound bacteria is 0.5-1.5%;

[0015] Preferably, by weight, the first compound bacteria includes: 30-50 parts of termite fungi, 30-50 parts of Trichoderma longipes, 30-50 parts of Phanerochaete chrysosporium, 30-50 parts of Aspergillus niger, and 10-30 parts of Bacillus subtilis.

[0016] Preferably, the effective viable count in the bacterial solution of the first compound bacteria is 1.5 to 2 × 10⁻⁶. 8 per g.

[0017] Preferably, the mass of the composite mycelium in step (3) is 0.5% to 2% of the mass of the pretreated substrate;

[0018] Preferably, by weight, the compound mycelium comprises: 10-30 parts of Russula rubra and 10-30 parts of Codonopsis pilosula;

[0019] Preferably, the culture temperature is 22–30°C, the humidity is 70–85%, with timely ventilation, and the CO2 concentration is ≤3.5 × 10⁻⁶. 3 ppm, culture time is 5-7 days.

[0020] Preferably, the inoculation amount of the second compound bacteria in step (4) is 1-2%;

[0021] Preferably, by weight, the second compound bacteria includes: 30-50 parts of brewer's yeast and 30-50 parts of bifidobacteria;

[0022] Preferably, during the second fermentation, the humidity of the fermentation material is maintained at 40-60%, the temperature at 37-40°C, and the fermentation time is 4-6 weeks;

[0023] Preferably, the effective viable count in the bacterial solution of the second compound bacteria is 0.5 to 1 × 10⁻⁶. 8 per g.

[0024] Preferably, the drying method in step (4) includes a flowing hot air drying method;

[0025] Preferably, the parameters of the flowing hot air drying method include: air temperature of 60-70°C and air flow rate of 1.5-3 m / s.

[0026] The present invention also provides a fermented straw feed protein additive prepared using the above preparation method.

[0027] This invention also provides the application of the above-mentioned fermented straw feed protein additive in improving the nutritional content of feed.

[0028] Preferably, the fermented straw feed protein additive is added at a mass of 0.5% to 2% of the base feed mass.

[0029] Beneficial Effects: This invention provides a method for preparing a fermented straw feed protein additive. Using dandelion and Jerusalem artichoke straw as base materials, the straw is steam-exploded and sterilized. It is then mixed with an organic nitrogen source and fermented using a first compound microorganism (termite fungus, Trichoderma longipes, Phanerochaete chrysosporium, Aspergillus niger, and Bacillus subtilis). The fermentation product is used as a culture medium for cultivating compound mycelia (Russula rubra and Codonopsis pilosula mycelia). After the mycelia have fully grown, a second compound microorganism (Saccharomyces cerevisiae and Bifidobacterium) is used for a second fermentation. The second fermentation product is dried and pulverized to obtain the fermented straw feed protein additive of this invention. This invention involves a first fermentation process that effectively degrades cellulose, hemicellulose, and lignin in straw, increasing its softness and expansion. Mycelial culture converts the urea produced during the degradation of cellulose, hemicellulose, and lignin into microbial protein, thereby increasing the total protein content of the material and producing microbial polysaccharides to provide energy for subsequent fermentation. The second fermentation process further degrades cellulose, hemicellulose, and lignin in straw into low-molecular-weight monosaccharides or oligosaccharides, converts insoluble carbohydrates in straw into soluble carbohydrates, and accumulates numerous metabolites such as amino acids, organic acids, alcohols, aldehydes, esters, and vitamins, improving the palatability of straw for feed and enhancing its preservative properties.

[0030] The preparation method described in this invention has the characteristics of low pollution, high efficiency, low cost, and suitability for industrial production. The fermented straw feed protein additive prepared by the method can have a protein content of over 20%, a crude fiber degradation rate of over 30%, and a phenolic acid content of over 7%. This solves the problems of insufficient utilization of agricultural straw and insignificant improvement in the digestibility, immunity, antioxidant capacity, and egg quality of laying hens in the prior art. Attached Figure Description

[0031] Figure 1 This is the LC-MS full spectrum identification of the metabolites from the first fermentation.

[0032] Figure 2This is the LC-MS full spectrum identification of the metabolites from the second fermentation. Detailed Implementation

[0033] This invention provides a method for preparing a fermented straw feed protein additive, comprising the following steps:

[0034] (1) Dandelion and Jerusalem artichoke stalks were crushed and then steam-exploded to obtain pretreated base material;

[0035] (2) After mixing the pretreated substrate described in step (1) with an organic nitrogen source, the first compound bacteria are inoculated to carry out the first fermentation to obtain the degraded material; the first compound bacteria include termite fungi, Trichoderma longipes, Phanerochaete chrysosporium, Aspergillus niger and Bacillus subtilis;

[0036] (3) After inoculating the degradation material described in step (2) with composite mycelium, culture is carried out to obtain a high mycelium protein material; the composite mycelium includes mycelium of Russula rubra and Codonopsis pilosula;

[0037] (4) The high microbial protein material described in step (3) is fermented using the second compound bacteria to obtain fermented high microbial protein material, which is then dried and pulverized to obtain the fermented straw feed protein additive; the second compound bacteria include brewer's yeast and Bifidobacterium.

[0038] This invention uses dandelion and Jerusalem artichoke stalks as raw materials. The dandelion and Jerusalem artichoke stalks are pulverized and then steam-exploded to obtain a pretreated substrate. The preferred mass ratio of dandelion to Jerusalem artichoke stalks in this invention is (0.1–1.5):(10–25), more preferably (0.7–1.2):(15–22), and most preferably 1:20. The dandelion used in this invention is preferably the whole plant. The dandelion and Jerusalem artichoke stalks are washed and dried (cut into 3–5 cm sections, first dehydrated by 20%–40% in a cool place, then sun-dried or baked in an oven at 60–75°C for 4–6 hours; avoid direct sunlight exposure of fresh dandelion and Jerusalem artichoke stalks), and then preferably pulverized. After pulverization, the pulverized material is passed through a 30-mesh sieve to obtain the pretreated material.

[0039] This invention preferably involves steam explosion of the pretreatment material, which is preferably performed in a high-temperature, high-pressure device using continuous high-temperature, high-pressure steam expansion explosion and sterilization. The high temperature is preferably ≥150℃, the high pressure is preferably ≥6MPa, the steam explosion time is preferably 15–30 min, and the air flow rate to pretreatment material volume-to-weight ratio is 2L:1kg. The material after steam explosion and sterilization is cooled to approximately 30℃ to obtain the pretreated substrate (also known as digestible sterilized substrate).

[0040] After obtaining the pretreated substrate, the present invention mixes the pretreated substrate with an organic nitrogen source and inoculates it with a first compound bacteria for a first fermentation to obtain degraded material; the first compound bacteria include termites, Trichoderma longipes, Phanerochaete chrysosporium, Aspergillus niger, and Bacillus subtilis. The organic nitrogen source of the present invention is 0.5-2% of the mass of the pretreated substrate, and the organic nitrogen source includes a compound organic nitrogen source, which, by mass parts, preferably includes: 5-10 parts yeast powder, 5-10 parts corn steep liquor powder, 5-10 parts soybean meal powder, and 5-10 parts peptone; more preferably includes 5-8 parts yeast powder, 7-10 parts corn steep liquor powder, 6-10 parts soybean meal powder, and 5-7 parts peptone; and most preferably includes 5 parts yeast powder, 10 parts corn steep liquor powder, 10 parts soybean meal powder, and 5 parts peptone.

[0041] This invention involves mixing the pretreated substrate and an organic nitrogen source, resulting in a mixture with a coefficient of variation (CV) ≤ 5.0% and a moisture content of 65%–70%. This invention uses this mixture as a fermentation substrate and conducts a first fermentation using a first compound microorganism. Preferably, the first compound microorganism is inoculated using a mixed bacterial solution, and the total viable count in the mixed bacterial solution is preferably 1.5–2 × 10⁻⁶. 8 / g. In this invention, the first compound bacteria preferably includes, by weight, 30-50 parts of termite fungi, 30-50 parts of Trichoderma longipes, 30-50 parts of Phanerochaete chrysosporium, 30-50 parts of Aspergillus niger, and 10-30 parts of Bacillus subtilis; more preferably, it includes 35-45 parts of termite fungi, 35-45 parts of Trichoderma longipes, 30-45 parts of Phanerochaete chrysosporium, 30-40 parts of Aspergillus niger, and 10-25 parts of Bacillus subtilis; and most preferably, it includes 40 parts of termite fungi, 40 parts of Trichoderma longipes, 40 parts of Phanerochaete chrysosporium, 30 parts of Aspergillus niger, and 15 parts of Bacillus subtilis.

[0042] In this invention, the mixture is thoroughly mixed after inoculation and then subjected to a first fermentation. During inoculation, the inoculation amount of the mixed bacterial solution is preferably 0.5–1.5%, more preferably 0.8–1.5%, and most preferably 1.2%. Thorough mixing is preferably defined as a coefficient of variation (CV) ≤ 5.0%. This invention preferably maintains the material moisture content at 40%–60% and the temperature at 37℃–40℃ for the first fermentation, with a fermentation time preferably of 5–7 days, effectively degrading cellulose, hemicellulose, and lignin in the straw to obtain degraded material.

[0043] After obtaining the degraded material, the present invention inoculates the degraded material with composite mycelia and then cultivates it to obtain a high-microbial-protein material; the composite mycelia include mycelia of Russula rubra and Codonopsis pilosula. When inoculating the composite mycelia, the present invention preferably maintains the degraded material under positive pressure and sterility. The mass of the composite mycelia is preferably 0.5-2% of the mass of the pretreated substrate, more preferably 1-1.5%, and most preferably 1.5%. By mass, the composite mycelia of the present invention preferably includes: 10-30 parts of Russula rubra and 10-30 parts of Codonopsis pilosula; more preferably, it includes: 10-15 parts of Russula rubra and 10-20 parts of Codonopsis pilosula; and most preferably, it includes: 10 parts of Russula rubra and 20 parts of Codonopsis pilosula.

[0044] In this invention, after inoculation, the mixture is thoroughly mixed, with a coefficient of variation (CV) ≤ 5.0%, and the temperature is controlled at 20–30°C. It is then placed in a sterile room for incubation, with the incubation temperature maintained between 22°C and 30°C, timely ventilation provided, humidity maintained between 70% and 85%, and CO2 concentration ≤ 3.5 × 10⁻⁶. 3 The culture time is 5-7 days, and a high microbial protein content is obtained. The culture method described in this invention can convert the nitrogen element produced during the first degradation of cellulose, hemicellulose and lignin into microbial protein, thereby increasing the total protein content of the material and producing microbial polysaccharides, providing energy for subsequent fermentation microorganisms.

[0045] After obtaining the high-microbial-protein material, the present invention utilizes a second compound bacteria to perform a second fermentation on the high-microbial-protein material to obtain a fermented high-microbial-protein material, which is then dried and pulverized to obtain the fermented straw feed protein additive. The second compound bacteria include brewer's yeast and Bifidobacterium. The inoculum amount of the second compound bacteria in the present invention is preferably 1-2%, more preferably 1-1.7%, and most preferably 1.5%. By weight, the second compound bacteria of the present invention preferably includes: 30-50 parts of brewer's yeast and 30-50 parts of Bifidobacterium; more preferably, it includes: 30-50 parts of brewer's yeast and 30-50 parts of Bifidobacterium; and most preferably, it includes: 50 parts of brewer's yeast and 35 parts of Bifidobacterium. The effective viable count in the bacterial solution of the second compound bacteria of the present invention is preferably 0.5-1×10⁻⁶. 8 The product is inoculated with the Dürr compound bacteria solution and mixed evenly. The coefficient of variation (CV) is ≤5.0%. The material moisture content is maintained at 40%–60%, and a temperature of 37℃–40℃ is used for a second fermentation for 4–6 weeks, yielding a fermented high-microbial-protein material. This second fermentation further degrades cellulose, hemicellulose, and lignin in straw into low-molecular-weight monosaccharides or oligosaccharides, converts insoluble carbohydrates in straw into soluble carbohydrates, and accumulates numerous metabolites such as amino acids, organic acids, alcohols, aldehydes, esters, and vitamins, improving the palatability of straw for feed and enhancing its preservative properties.

[0046] After obtaining the fermented high-microbial-protein material, the present invention preferably further includes drying, which preferably includes hot air drying with parameters including: air temperature 60℃~70℃, air velocity 1.5~3m / s. After drying, the present invention preferably further includes pulverization and packaging to obtain the fermented straw feed protein additive (hereinafter referred to as feed additive). The pulverization in the present invention is preferably pulverized to 80~100 mesh, and then packaged into feed additive.

[0047] The present invention also provides a fermented straw feed protein additive prepared using the above preparation method.

[0048] The feed additive described in this invention has an average crude protein content of up to 20.49%, a crude fiber degradation rate of over 30%, and an average phenolic acid content of up to 7.46%, making it rich in nutrients.

[0049] This invention also provides the application of the above-mentioned fermented straw feed protein additive in improving the nutritional content of feed.

[0050] The feed additive of the present invention can be mixed with feed, that is, the feed additive is applied to feed, and the added mass of the fermented straw feed protein additive is preferably 0.5 to 2% of the mass of the basic feed.

[0051] To further illustrate the present invention, the following detailed description, in conjunction with embodiments, provides a fermented straw feed protein additive, its preparation method, and its application. However, these descriptions should not be construed as limiting the scope of protection of the present invention.

[0052] Example 1

[0053] (1) Digestion and sterilization substrate.

[0054] Dandelion (whole plant, 1 kg) and Jerusalem artichoke stalks (20 kg) were washed, dried, crushed, and passed through a 30-mesh sieve to obtain pretreatment material. The pretreatment material was then placed in a high-temperature, high-pressure device for continuous high-temperature, high-pressure steam expansion, explosion, and sterilization (temperature: 180℃, pressure: 10MPa, duration: 30 min, air volume to pretreatment material volume weight ratio: 2L:1kg). After sterilization, the material was cooled to approximately 30℃.

[0055] (2) Mixed materials

[0056] Take all the digestible sterilized base material and add 0.42 kg of compound organic nitrogen source (28.6% yeast powder, 14.2% corn steep liquor powder, 28.6% soybean meal powder, and 28.6% peptone), and mix evenly. The coefficient of variation (CV) should be ≤5.0%, and the moisture content should be 65%–70%.

[0057] (3) Degradable materials

[0058] Add 0.32 kg of compound bacteria (termite fungi 15.8%, Trichoderma longipes 26.3%, Phanerochaete chrysosporium 26.3%, Aspergillus niger 18.4%, Bacillus subtilis 13.2%) to the mixed materials, resulting in an effective viable count of 1.5–2 × 10⁻⁶ bacteria. 8 The mixture should be prepared in batches of 100g each, with a coefficient of variation (CV) ≤ 5.0%. The material should be kept at a moisture content of 40%–60% and the temperature of 37℃–40℃ for the first fermentation, which should last for 6 days.

[0059] Example 2

[0060] Steps (1), (2), and (3) are the same as in Example 1.

[0061] (4) High bacterial protein materials

[0062] Take the decomposed material and inoculate it with 0.43 kg of compound mycelium (33.3% Russula rubra and 66.7% Codonopsis pilosula mycelium) under positive pressure and aseptic conditions, mixing thoroughly. The coefficient of variation (CV) should be ≤5.0%, and the temperature should be controlled between 20 and 30℃. After inoculation, place it in a sterile room for cultivation, maintaining the temperature between 22℃ and 30℃ with timely ventilation, humidity between 70% and 85%, and CO2 concentration ≤3.5×10⁻⁶. 3 ppm, culture time 6 days.

[0063] (5) Fermented high microbial protein materials

[0064] Take high mycelial protein material, add 0.44 kg of compound bacteria (58.8% brewer's yeast and 41.2% bifidobacteria), mix evenly, with a coefficient of variation CV ≤ 5.0%, maintain material moisture at 40%–60%, and carry out a second fermentation at 37℃–40℃ for 5 weeks.

[0065] The first fermentation material from Example 1 and the second fermentation material from Example 2 were used for qualitative and quantitative analysis of the first and second fermentation metabolites by chromatography-mass spectrometry (LC-MS). Figure 1 , Figure 2 ).

[0066] Example 3

[0067] (1) Digestion and sterilization substrate.

[0068] Dandelion (whole plant, 1 kg) and Jerusalem artichoke stalks (20 kg) were washed, dried, crushed, and passed through a 30-mesh sieve to obtain pretreatment material. The pretreatment material was then placed in a high-temperature, high-pressure device for continuous high-temperature, high-pressure steam expansion, explosion, and sterilization (temperature: 180℃, pressure: 10MPa, duration: 30 min, air volume to pretreatment material volume weight ratio: 2L:1kg). After sterilization, the material was cooled to approximately 30℃.

[0069] (2) Mixed materials

[0070] Take all the digestible sterilized base material and add 0.42 kg of compound organic nitrogen source (28.6% yeast powder, 14.2% corn steep liquor powder, 28.6% soybean meal powder, and 28.6% peptone), and mix evenly. The coefficient of variation (CV) should be ≤5.0%, and the moisture content should be 65%–70%.

[0071] (3) Degradable materials

[0072] Take the mixture and add 0.32 kg of compound bacteria (15.8% termite fungus, 26.3% Trichoderma longipes, 26.3% Phanerochaete chrysosporium, 18.4% Aspergillus niger, and 13.2% Bacillus subtilis), with an effective viable count of 1.5 to 2 × 10⁸ CFU / g. Mix well, with a coefficient of variation (CV) ≤ 5.0%. Maintain the material moisture content at 40% to 60% and the temperature at 37℃ to 40℃ for the first fermentation, which takes 6 days.

[0073] (4) High bacterial protein materials

[0074] Take the decomposed material and inoculate it with 0.2 kg of compound mycelium (33.3% Russula rubra and 66.7% Codonopsis pilosula mycelium) under positive pressure and aseptic conditions, mixing thoroughly. The coefficient of variation (CV) should be ≤5.0%, and the temperature should be controlled between 20 and 30℃. After inoculation, place it in a sterile room for cultivation, maintaining the temperature between 22℃ and 30℃ with timely ventilation, humidity between 70% and 85%, and CO2 concentration ≤3.5×10⁻⁶. 3 ppm, culture time 6 days.

[0075] Example 4

[0076] Steps (1), (2), and (3) are the same as in Example 3.

[0077] (4) High bacterial protein materials

[0078] Take the decomposed material and inoculate it with 0.4 kg of compound mycelium (33.3% Russula rubra and 66.7% Codonopsis pilosula mycelium) under positive pressure and aseptic conditions, mixing thoroughly. The coefficient of variation (CV) should be ≤5.0%, and the temperature should be controlled between 20 and 30℃. After inoculation, place it in a sterile room for cultivation, maintaining the temperature between 22℃ and 30℃ with timely ventilation, humidity between 70% and 85%, and CO2 concentration ≤3.5×10⁻⁶. 3 ppm, culture time 6 days.

[0079] Example 5

[0080] Steps (1), (2), and (3) are the same as in Example 3.

[0081] (4) High bacterial protein materials

[0082] Take the decomposed material and inoculate it with 0.6 kg of compound mycelium (33.3% Russula rubra and 66.7% Codonopsis pilosula mycelium) under positive pressure and aseptic conditions, mixing thoroughly. The coefficient of variation (CV) should be ≤5.0%, and the temperature should be controlled between 20 and 30℃. After inoculation, place it in a sterile room for cultivation, maintaining the cultivation temperature between 22℃ and 30℃ with timely ventilation, humidity between 70% and 85%, and CO2 concentration ≤3.5×10⁻⁶. 3 ppm, culture time 6 days.

[0083] Example 6

[0084] Steps (1), (2), and (3) are the same as in Example 3.

[0085] (4) High bacterial protein materials

[0086] Take the decomposed material and inoculate it with 0.8 kg of compound mycelium (33.3% Russula rubra and 66.7% Codonopsis pilosula mycelium) under positive pressure and aseptic conditions, mixing thoroughly. The coefficient of variation (CV) should be ≤5.0%, and the temperature should be controlled between 20 and 30℃. After inoculation, place it in a sterile room for cultivation, maintaining the temperature between 22℃ and 30℃ with timely ventilation, humidity between 70% and 85%, and CO2 concentration ≤3.5×10⁻⁶. 3 ppm, culture time 6 days.

[0087] Example 7

[0088] Steps (1), (2), and (3) are the same as in Example 3.

[0089] (4) High bacterial protein materials

[0090] Take the decomposed material and inoculate it with 1.0 kg of compound mycelium (33.3% Russula rubra and 66.7% Codonopsis pilosula mycelium) under positive pressure and aseptic conditions, mixing thoroughly. The coefficient of variation (CV) should be ≤5.0%, and the temperature should be controlled between 20 and 30℃. After inoculation, place it in a sterile room for cultivation, maintaining the cultivation temperature between 22℃ and 30℃ with timely ventilation, humidity between 70% and 85%, and CO2 concentration ≤3.5×10⁻⁶. 3 ppm, culture time 6 days.

[0091] Comparative Example 1

[0092] (1) Digestion and sterilization substrate.

[0093] Dandelion (whole plant, 1 kg) and Jerusalem artichoke stalks (20 kg) were washed, dried, crushed, and passed through a 30-mesh sieve to obtain pretreatment material. The pretreatment material was then placed in a high-temperature, high-pressure device for continuous high-temperature, high-pressure steam expansion, explosion, and sterilization (temperature: 180℃, pressure: 10MPa, duration: 30 min, air volume to pretreatment material volume weight ratio: 2L:1kg). After sterilization, the material was cooled to approximately 30℃.

[0094] (2) Mixed materials

[0095] Take all the digestible sterilized base material and add 0.42 kg of compound organic nitrogen source (28.6% yeast powder, 14.2% corn steep liquor powder, 28.6% soybean meal powder, and 28.6% peptone), and mix evenly. The coefficient of variation (CV) should be ≤5.0%, and the moisture content should be 65%–70%.

[0096] (3) Degradable materials

[0097] Add 0.32 kg of compound bacteria (termite fungi 15.8%, Trichoderma longipes 26.3%, Phanerochaete chrysosporium 26.3%, Aspergillus niger 18.4%, Bacillus subtilis 13.2%) to the mixed materials, resulting in an effective viable count of 1.5–2 × 10⁻⁶ bacteria. 8 The mixture should be prepared in batches of 100g each, with a coefficient of variation (CV) ≤ 5.0%. The material should be kept at a moisture content of 40%–60% and the temperature of 37℃–40℃ for the first fermentation, which should last for 6 days.

[0098] The high-protein microbial materials from Examples 3-7 and the first fermentation material from Comparative Example 1 were used to determine the crude protein content of the microbial cells.

[0099] The crude protein content of the bacterial cells was determined according to the method in GB / T 6432-2018.

[0100] Table 1. Crude protein, crude fiber, and phenolic acid content of straw materials treated under different embodiment and comparative conditions.

[0101] Comparative Example 1 Example 3 Example 4 Example 5 Example 6 Example 7 Bacterial protein (%) 8.51％ 15.40％ 18.65％ 16.33％ 16.59％ 14.78％

[0102] Example 8

[0103] (1) Digestion and sterilization substrate.

[0104] Dandelion (whole plant, 1 kg) and Jerusalem artichoke stalks (20 kg) were washed, dried, crushed, and passed through a 30-mesh sieve to obtain pre-treatment material. The pre-treatment material was then placed in a high-temperature, high-pressure device for continuous high-temperature, high-pressure steam expansion, explosion, and sterilization (temperature: 150℃, pressure: 6 MPa, duration: 15 min, air volume to pre-treatment material volume weight ratio: 2 L: 1 kg). After sterilization, the material was cooled to approximately 30℃.

[0105] (2) Mixed materials

[0106] Take all the digestible sterilized base material and add 0.11 kg of compound organic nitrogen source (16.7% yeast powder, 16.7% corn steep liquor powder, 33.3% soybean meal powder, 33.3% peptone), and mix evenly. The coefficient of variation (CV) should be ≤5.0%, and the moisture content should be 65%–70%.

[0107] (3) Degradable materials

[0108] Add 0.13 kg of compound bacteria (termite fungi 18.2%, Trichoderma longipes 27.3%, Phanerochaete chrysosporium 24.2%, Aspergillus niger 24.2%, Bacillus subtilis 6.1%) to the mixed materials, resulting in a total effective viable count of 1.5–2 × 10⁻⁶. 8 The sample is prepared by mixing the sample with a density of 1 / g and ensuring uniform mixing. The coefficient of variation (CV) is ≤5.0%. The material is kept at a moisture content of 40%–60% and the temperature is 37℃–40℃ for the first fermentation, which lasts for 5 days.

[0109] (4) High bacterial protein materials

[0110] Take the decomposed material and inoculate it with 0.11 kg of compound mycelium (50% Russula rubra and 50% Codonopsis pilosula mycelium) under positive pressure and sterile conditions, mixing thoroughly. The coefficient of variation (CV) should be ≤5.0%, and the temperature should be controlled between 20 and 30℃. After inoculation, place it in a sterile room for cultivation, maintaining the cultivation temperature between 22℃ and 30℃ with timely ventilation, humidity between 70% and 85%, and CO2 concentration ≤3.5×10⁻⁶. 3 ppm, culture time 5 days.

[0111] (5) Fermented high microbial protein materials

[0112] Take high mycelial protein material, add 0.21 kg of compound bacteria (50% brewer's yeast and 50% bifidobacteria), mix evenly, with a coefficient of variation CV ≤ 5.0%, maintain material moisture at 40%–60%, and carry out a second fermentation at 37℃–40℃ for 4 weeks.

[0113] Example 9

[0114] (1) Digestion and sterilization substrate.

[0115] Dandelion (whole plant, 1 kg) and Jerusalem artichoke stalks (20 kg) were washed, dried, crushed, and passed through a 30-mesh sieve to obtain pretreatment material. The pretreatment material was then placed in a high-temperature, high-pressure device for continuous high-temperature, high-pressure steam expansion, explosion, and sterilization (temperature: 160℃, pressure: 8 MPa, duration: 20 min, air volume to pretreatment material volume weight ratio: 2 L: 1 kg). After sterilization, the material was cooled to approximately 30℃.

[0116] (2) Mixed materials

[0117] Take all the digestible sterilized base material and add 0.21 kg of compound organic nitrogen source (16.7% yeast powder, 33.3% corn steep liquor powder, 33.3% soybean meal powder, 16.7% peptone), mix evenly, coefficient of variation CV ≤ 5.0%, moisture content 65%~70%.

[0118] (3) Degradable materials

[0119] Add 0.19 kg of compound bacteria (termite fungi 24.2%, Trichoderma longipes 24.2%, Phanerochaete chrysosporium 24.2%, Aspergillus niger 18.2%, Bacillus subtilis 9.1%) to the mixed materials, resulting in an effective viable count of 1.5–2 × 10⁻⁶ bacteria. 8 The sample is prepared by mixing the sample with a density of 1 / g and ensuring uniform mixing. The coefficient of variation (CV) is ≤5.0%. The material is kept at a moisture content of 40%–60% and the temperature is 37℃–40℃ for the first fermentation, which lasts for 5 days.

[0120] (4) High bacterial protein materials

[0121] Take the decomposed material and inoculate it with 0.21 kg of compound mycelium (42.9% Russula rubra and 57.1% Codonopsis pilosula mycelium) under positive pressure and aseptic conditions, mixing thoroughly. The coefficient of variation (CV) should be ≤5.0%, and the temperature should be controlled between 20 and 30℃. After inoculation, place it in a sterile room for cultivation, maintaining the temperature between 22℃ and 30℃ with timely ventilation, humidity between 70% and 85%, and CO2 concentration ≤3.5×10⁻⁶. 3 ppm, culture time 5 days.

[0122] (5) Fermented high microbial protein materials

[0123] Take high mycelial protein material, add 0.30 kg of compound bacteria (46.7% brewer's yeast and 53.3% bifidobacteria), mix evenly, with a coefficient of variation CV ≤ 5.0%, maintain material moisture at 40%–60%, and carry out a second fermentation at 37℃–40℃ for 4 weeks.

[0124] Example 10

[0125] (1) Digestion and sterilization substrate.

[0126] Dandelion (whole plant, 1 kg) and Jerusalem artichoke stalks (20 kg) were washed, dried, crushed, and passed through a 30-mesh sieve to obtain pretreatment material. The pretreatment material was then placed in a high-temperature, high-pressure device for continuous high-temperature, high-pressure steam expansion, explosion, and sterilization (temperature: 170℃, pressure: 9 MPa, duration: 25 min, air volume to pretreatment material volume weight ratio: 2 L: 1 kg). After sterilization, the material was cooled to approximately 30℃.

[0127] (2) Mixed materials

[0128] Take all the digestible sterilized base material and add 0.32 kg of compound organic nitrogen source (33.3% yeast powder, 16.7% corn steep liquor powder, 16.7% soybean meal powder, and 33.3% peptone), and mix evenly. The coefficient of variation (CV) should be ≤5.0%, and the moisture content should be 65%–70%.

[0129] (3) Degradable materials

[0130] Add 0.26 kg of compound bacteria (18.4% termite fungi, 18.4% Trichoderma longipetiae, 26.3% Phanerochaete chrysosporium, 21.1% Aspergillus niger, and 15.8% Bacillus subtilis) to the mixed materials, resulting in an effective viable count of 1.5–2 × 10⁻⁶ bacteria. 8 The sample is prepared by mixing the sample with a density of 1 / g and ensuring uniform mixing. The coefficient of variation (CV) is ≤5.0%. The material is kept at a moisture content of 40%–60% and the temperature is 37℃–40℃ for the first fermentation, which lasts for 5 days.

[0131] (4) High bacterial protein materials

[0132] Take the decomposed material and inoculate it with 0.32 kg of compound mycelium (60% Russula rubra and 40% Codonopsis pilosula mycelium) under positive pressure and aseptic conditions, mixing thoroughly. The coefficient of variation (CV) should be ≤5.0%, and the temperature should be controlled between 20 and 30℃. After inoculation, place it in a sterile room for cultivation, maintaining the cultivation temperature between 22℃ and 30℃ with timely ventilation, humidity between 70% and 85%, and CO2 concentration ≤3.5×10⁻⁶. 3 ppm, culture time 5 days.

[0133] (5) Fermented high microbial protein materials

[0134] Take high mycelial protein material, add 0.37 kg of compound bacteria (44.4% brewer's yeast and 55.5% bifidobacteria), mix evenly, with a coefficient of variation CV ≤ 5.0%, maintain material moisture at 40%–60%, and carry out a second fermentation at 37℃–40℃ for 4 weeks.

[0135] Example 11

[0136] (1) Digestion and sterilization substrate.

[0137] Dandelion (whole plant, 1 kg) and Jerusalem artichoke stalks (20 kg) were washed, dried, crushed, and passed through a 30-mesh sieve to obtain pretreatment material. The pretreatment material was then placed in a high-temperature, high-pressure device for continuous high-temperature, high-pressure steam expansion, explosion, and sterilization (temperature: 180℃, pressure: 10MPa, duration: 30 min, air volume to pretreatment material volume weight ratio: 2L:1kg). After sterilization, the material was cooled to approximately 30℃.

[0138] (2) Mixed materials

[0139] Take all the digestible sterilized base material and add 0.42 kg of compound organic nitrogen source (28.6% yeast powder, 14.2% corn steep liquor powder, 28.6% soybean meal powder, and 28.6% peptone), and mix evenly. The coefficient of variation (CV) should be ≤5.0%, and the moisture content should be 65%–70%.

[0140] (3) Degradable materials

[0141] Add 0.32 kg of compound bacteria (termite fungi 15.8%, Trichoderma longipes 26.3%, Phanerochaete chrysosporium 26.3%, Aspergillus niger 18.4%, Bacillus subtilis 13.2%) to the mixed materials, resulting in an effective viable count of 1.5–2 × 10⁻⁶ bacteria. 8 The sample is prepared by mixing the sample with a density of 1 / g and ensuring uniform mixing. The coefficient of variation (CV) is ≤5.0%. The material is kept at a moisture content of 40%–60% and the temperature is 37℃–40℃ for the first fermentation, which lasts for 5 days.

[0142] (4) High bacterial protein materials

[0143] Take the decomposed material and inoculate it with 0.43 kg of compound mycelium (33.3% Russula rubra and 66.7% Codonopsis pilosula mycelium) under positive pressure and aseptic conditions, mixing thoroughly. The coefficient of variation (CV) should be ≤5.0%, and the temperature should be controlled between 20 and 30℃. After inoculation, place it in a sterile room for cultivation, maintaining the temperature between 22℃ and 30℃ with timely ventilation, humidity between 70% and 85%, and CO2 concentration ≤3.5×10⁻⁶. 3 ppm, culture time 5 days.

[0144] (5) Fermented high microbial protein materials

[0145] Take high mycelial protein material, add 0.44 kg of compound bacteria (58.8% brewer's yeast and 41.2% bifidobacteria), mix evenly, with a coefficient of variation CV ≤ 5.0%, maintain material moisture at 40%–60%, and carry out a second fermentation at 37℃–40℃ for 4 weeks.

[0146] Example 12

[0147] (1) Digestion and sterilization substrate.

[0148] Dandelion (whole plant, 1 kg) and Jerusalem artichoke stalks (20 kg) were washed, dried, crushed, and passed through a 30-mesh sieve to obtain pre-treatment material. The pre-treatment material was then placed in a high-temperature, high-pressure device for continuous high-temperature, high-pressure steam expansion, explosion, and sterilization (temperature: 150℃, pressure: 6 MPa, duration: 15 min, air volume to pre-treatment material volume weight ratio: 2 L: 1 kg). After sterilization, the material was cooled to approximately 30℃.

[0149] (2) Mixed materials

[0150] Take all the digestible sterilized base material and add 0.11 kg of compound organic nitrogen source (16.7% yeast powder, 16.7% corn steep liquor powder, 33.3% soybean meal powder, 33.3% peptone), and mix evenly. The coefficient of variation (CV) should be ≤5.0%, and the moisture content should be 65%–70%.

[0151] (3) Degradable materials

[0152] Add 0.13 kg of compound bacteria (termite fungi 18.2%, Trichoderma longipes 27.3%, Phanerochaete chrysosporium 24.2%, Aspergillus niger 24.2%, Bacillus subtilis 6.1%) to the mixed materials, resulting in an effective viable count of 1.5–2 × 10⁻⁶ bacteria. 8 The mixture should be prepared in batches of 100g each, with a coefficient of variation (CV) ≤ 5.0%. The material should be kept at a moisture content of 40%–60% and the temperature of 37℃–40℃ for the first fermentation, which should last for 6 days.

[0153] (4) High bacterial protein materials

[0154] Take the decomposed material and inoculate it with 0.11 kg of compound mycelium (50% Russula rubra and 50% Codonopsis pilosula mycelium) under positive pressure and sterile conditions, mixing thoroughly. The coefficient of variation (CV) should be ≤5.0%, and the temperature should be controlled between 20 and 30℃. After inoculation, place it in a sterile room for cultivation, maintaining the cultivation temperature between 22℃ and 30℃ with timely ventilation, humidity between 70% and 85%, and CO2 concentration ≤3.5×103ppm for 6 days.

[0155] (5) Fermented high microbial protein materials

[0156] Take high mycelial protein material, add 0.21 kg of compound bacteria (50% brewer's yeast and 50% bifidobacteria), mix evenly, with a coefficient of variation (CV) ≤ 5.0%, maintain material moisture at 40%–60%, and carry out a second fermentation at 37℃–40℃ for 5 weeks.

[0157] Example 13

[0158] (1) Digestion and sterilization substrate.

[0159] Dandelion (whole plant, 1 kg) and Jerusalem artichoke stalks (20 kg) were washed, dried, crushed, and passed through a 30-mesh sieve to obtain pretreatment material. The pretreatment material was then placed in a high-temperature, high-pressure device for continuous high-temperature, high-pressure steam expansion, explosion, and sterilization (temperature: 160℃, pressure: 8 MPa, duration: 20 min, air volume to pretreatment material volume weight ratio: 2 L: 1 kg). After sterilization, the material was cooled to approximately 30℃.

[0160] (2) Mixed materials

[0161] Take all the digestible sterilized base material and add 0.21 kg of compound organic nitrogen source (16.7% yeast powder, 33.3% corn steep liquor powder, 33.3% soybean meal powder, 16.7% peptone), mix evenly, coefficient of variation CV ≤ 5.0%, moisture content 65%~70%.

[0162] (3) Degradable materials

[0163] Add 0.19 kg of compound bacteria (termite fungi 24.2%, Trichoderma longipes 24.2%, Phanerochaete chrysosporium 24.2%, Aspergillus niger 18.2%, Bacillus subtilis 9.1%) to the mixed materials, resulting in an effective viable count of 1.5–2 × 10⁻⁶ bacteria. 8 The mixture should be prepared in batches of 100g each, with a coefficient of variation (CV) ≤ 5.0%. The material should be kept at a moisture content of 40%–60% and the temperature of 37℃–40℃ for the first fermentation, which should last for 6 days.

[0164] (4) High bacterial protein materials

[0165] Take the decomposed material and inoculate it with 0.21 kg of compound mycelium (42.9% Russula rubra and 57.1% Codonopsis pilosula mycelium) under positive pressure and aseptic conditions, mixing thoroughly. The coefficient of variation (CV) should be ≤5.0%, and the temperature should be controlled between 20 and 30℃. After inoculation, place it in a sterile room for cultivation, maintaining the temperature between 22℃ and 30℃ with timely ventilation, humidity between 70% and 85%, and CO2 concentration ≤3.5×10⁻⁶. 3 ppm, culture time 6 days.

[0166] (5) Fermented high microbial protein materials

[0167] Take high mycelial protein material, add 0.30 kg of compound bacteria (46.7% brewer's yeast and 53.3% bifidobacteria), mix evenly, with a coefficient of variation CV ≤ 5.0%, maintain material moisture at 40%–60%, and carry out a second fermentation at 37℃–40℃ for 5 weeks.

[0168] Example 14

[0169] (1) Digestion and sterilization substrate.

[0170] Dandelion (whole plant, 1 kg) and Jerusalem artichoke stalks (20 kg) were washed, dried, crushed, and passed through a 30-mesh sieve to obtain pretreatment material. The pretreatment material was then placed in a high-temperature, high-pressure device for continuous high-temperature, high-pressure steam expansion, explosion, and sterilization (temperature: 170℃, pressure: 9 MPa, duration: 25 min, air volume to pretreatment material volume weight ratio: 2 L: 1 kg). After sterilization, the material was cooled to approximately 30℃.

[0171] (2) Mixed materials

[0172] Take all the digestible sterilized base material and add 0.32 kg of compound organic nitrogen source (33.3% yeast powder, 16.7% corn steep liquor powder, 16.7% soybean meal powder, and 33.3% peptone), and mix evenly. The coefficient of variation (CV) should be ≤5.0%, and the moisture content should be 65%–70%.

[0173] (3) Degradable materials

[0174] Add 0.26 kg of compound bacteria (18.4% termite fungi, 18.4% Trichoderma longipetiae, 26.3% Phanerochaete chrysosporium, 21.1% Aspergillus niger, and 15.8% Bacillus subtilis) to the mixed materials, resulting in an effective viable count of 1.5–2 × 10⁻⁶ bacteria. 8 The mixture should be prepared in batches of 100g each, with a coefficient of variation (CV) ≤ 5.0%. The material should be kept at a moisture content of 40%–60% and the temperature of 37℃–40℃ for the first fermentation, which should last for 6 days.

[0175] (4) High bacterial protein materials

[0176] Take the decomposed material and inoculate it with 0.32 kg of compound mycelium (60% Russula rubra and 40% Codonopsis pilosula mycelium) under positive pressure and aseptic conditions, mixing thoroughly. The coefficient of variation (CV) should be ≤5.0%, and the temperature should be controlled between 20 and 30℃. After inoculation, place it in a sterile room for cultivation, maintaining the cultivation temperature between 22℃ and 30℃ with timely ventilation, humidity between 70% and 85%, and CO2 concentration ≤3.5×10⁻⁶. 3 ppm, culture time 6 days.

[0177] (5) Fermented high microbial protein materials

[0178] Take high mycelial protein material, add 0.37 kg of compound bacteria (44.4% brewer's yeast and 55.5% bifidobacteria), mix evenly, with a coefficient of variation CV ≤ 5.0%, maintain material moisture at 40%–60%, and carry out a second fermentation at 37℃–40℃ for 5 weeks.

[0179] Example 15

[0180] (1) Digestion and sterilization substrate.

[0181] Dandelion (whole plant, 1 kg) and Jerusalem artichoke stalks (20 kg) were washed, dried, crushed, and passed through a 30-mesh sieve to obtain pretreatment material. The pretreatment material was then placed in a high-temperature, high-pressure device for continuous high-temperature, high-pressure steam expansion, explosion, and sterilization (temperature: 180℃, pressure: 10MPa, duration: 30 min, air volume to pretreatment material volume weight ratio: 2L:1kg). After sterilization, the material was cooled to approximately 30℃.

[0182] (2) Mixed materials

[0183] Take all the digestible sterilized base material and add 0.42 kg of compound organic nitrogen source (28.6% yeast powder, 14.2% corn steep liquor powder, 28.6% soybean meal powder, and 28.6% peptone), and mix evenly. The coefficient of variation (CV) should be ≤5.0%, and the moisture content should be 65%–70%.

[0184] (3) Degradable materials

[0185] Add 0.32 kg of compound bacteria (termite fungi 15.8%, Trichoderma longipes 26.3%, Phanerochaete chrysosporium 26.3%, Aspergillus niger 18.4%, Bacillus subtilis 13.2%) to the mixed materials, resulting in an effective viable count of 1.5–2 × 10⁻⁶ bacteria. 8 The mixture should be prepared in batches of 100g each, with a coefficient of variation (CV) ≤ 5.0%. The material should be kept at a moisture content of 40%–60% and the temperature of 37℃–40℃ for the first fermentation, which should last for 6 days.

[0186] (4) High bacterial protein materials

[0187] Take the decomposed material and inoculate it with 0.43 kg of compound mycelium (33.3% Russula rubra and 66.7% Codonopsis pilosula mycelium) under positive pressure and aseptic conditions, mixing thoroughly. The coefficient of variation (CV) should be ≤5.0%, and the temperature should be controlled between 20 and 30℃. After inoculation, place it in a sterile room for cultivation, maintaining the temperature between 22℃ and 30℃ with timely ventilation, humidity between 70% and 85%, and CO2 concentration ≤3.5×10⁻⁶. 3 ppm, culture time 6 days.

[0188] (5) Fermented high microbial protein materials

[0189] Take high mycelial protein material, add 0.44 kg of compound bacteria (58.8% brewer's yeast and 41.2% bifidobacteria), mix evenly, with a coefficient of variation CV ≤ 5.0%, maintain material moisture at 40%–60%, and carry out a second fermentation at 37℃–40℃ for 5 weeks.

[0190] Example 16

[0191] (1) Digestion and sterilization substrate.

[0192] Dandelion (whole plant, 1 kg) and Jerusalem artichoke stalks (20 kg) were washed, dried, crushed, and passed through a 30-mesh sieve to obtain pre-treatment material. The pre-treatment material was then placed in a high-temperature, high-pressure device for continuous high-temperature, high-pressure steam expansion, explosion, and sterilization (temperature: 150℃, pressure: 6 MPa, duration: 15 min, air volume to pre-treatment material volume weight ratio: 2 L: 1 kg). After sterilization, the material was cooled to approximately 30℃.

[0193] (2) Mixed materials

[0194] Take all the digestible sterilized base material and add 0.11 kg of compound organic nitrogen source (16.7% yeast powder, 16.7% corn steep liquor powder, 33.3% soybean meal powder, 33.3% peptone), and mix evenly. The coefficient of variation (CV) should be ≤5.0%, and the moisture content should be 65%–70%.

[0195] (3) Degradable materials

[0196] Add 0.13 kg of compound bacteria (termite fungi 18.2%, Trichoderma longipes 27.3%, Phanerochaete chrysosporium 24.2%, Aspergillus niger 24.2%, Bacillus subtilis 6.1%) to the mixed materials, resulting in an effective viable count of 1.5–2 × 10⁻⁶ bacteria. 8 The mixture should be prepared in batches of 100g each, with a coefficient of variation (CV) ≤ 5.0%. The material should be kept at a moisture content of 40%–60% and the temperature of 37℃–40℃ for the first fermentation, which should last for 7 days.

[0197] (4) High bacterial protein materials

[0198] Take the decomposed material and inoculate it with 0.11 kg of compound mycelium (50% Russula rubra and 50% Codonopsis pilosula mycelium) under positive pressure and sterile conditions, mixing thoroughly. The coefficient of variation (CV) should be ≤5.0%, and the temperature should be controlled between 20 and 30℃. After inoculation, place it in a sterile room for cultivation, maintaining the cultivation temperature between 22℃ and 30℃ with timely ventilation, humidity between 70% and 85%, and CO2 concentration ≤3.5×10⁻⁶. 3 ppm, culture time 7 days.

[0199] (5) Fermented high microbial protein materials

[0200] Take high mycelial protein material, add 0.21 kg of compound bacteria (50% brewer's yeast and 50% bifidobacteria), mix evenly, with a coefficient of variation CV ≤ 5.0%, maintain material moisture at 40%–60%, and carry out a second fermentation at 37℃–40℃ for 6 weeks.

[0201] Example 17

[0202] (1) Digestion and sterilization substrate.

[0203] Dandelion (whole plant, 1 kg) and Jerusalem artichoke stalks (20 kg) were washed, dried, crushed, and passed through a 30-mesh sieve to obtain pretreatment material. The pretreatment material was then placed in a high-temperature, high-pressure device for continuous high-temperature, high-pressure steam expansion, explosion, and sterilization (temperature: 160℃, pressure: 8 MPa, duration: 20 min, air volume to pretreatment material volume weight ratio: 2 L: 1 kg). After sterilization, the material was cooled to approximately 30℃.

[0204] (2) Mixed materials

[0205] Take all the digestible sterilized base material and add 0.21 kg of compound organic nitrogen source (16.7% yeast powder, 33.3% corn steep liquor powder, 33.3% soybean meal powder, 16.7% peptone), mix evenly, coefficient of variation CV ≤ 5.0%, moisture content 65%~70%.

[0206] (3) Degradable materials

[0207] Add 0.19 kg of compound bacteria (termite fungi 24.2%, Trichoderma longipes 24.2%, Phanerochaete chrysosporium 24.2%, Aspergillus niger 18.2%, Bacillus subtilis 9.1%) to the mixed materials, resulting in an effective viable count of 1.5–2 × 10⁻⁶ bacteria. 8 The mixture should be prepared in batches of 100g each, with a coefficient of variation (CV) ≤ 5.0%. The material should be kept at a moisture content of 40%–60% and the temperature of 37℃–40℃ for the first fermentation, which should last for 7 days.

[0208] (4) High bacterial protein materials

[0209] Take the decomposed material and inoculate it with 0.21 kg of compound mycelium (42.9% Russula rubra and 57.1% Codonopsis pilosula mycelium) under positive pressure and aseptic conditions, mixing thoroughly. The coefficient of variation (CV) should be ≤5.0%, and the temperature should be controlled between 20 and 30℃. After inoculation, place it in a sterile room for cultivation, maintaining the temperature between 22℃ and 30℃ with timely ventilation, humidity between 70% and 85%, and CO2 concentration ≤3.5×10⁻⁶. 3 ppm, culture time 7 days.

[0210] (5) Fermented high microbial protein materials

[0211] Take high mycelial protein material, add 0.30 kg of compound bacteria (46.7% brewer's yeast and 57.1% bifidobacteria), mix evenly, with a coefficient of variation CV ≤ 5.0%, maintain material moisture at 40%–60%, and carry out a second fermentation at 37℃–40℃ for 6 weeks.

[0212] Example 18

[0213] (1) Digestion and sterilization substrate.

[0214] Dandelion (whole plant, 1 kg) and Jerusalem artichoke stalks (20 kg) were washed, dried, crushed, and passed through a 30-mesh sieve to obtain pretreatment material. The pretreatment material was then placed in a high-temperature, high-pressure device for continuous high-temperature, high-pressure steam expansion, explosion, and sterilization (temperature: 170℃, pressure: 9 MPa, duration: 25 min, air volume to pretreatment material volume weight ratio: 2 L: 1 kg). After sterilization, the material was cooled to approximately 30℃.

[0215] (2) Mixed materials

[0216] Take all the digestible sterilized base material and add 0.32 kg of compound organic nitrogen source (33.3% yeast powder, 16.7% corn steep liquor powder, 16.7% soybean meal powder, and 33.3% peptone), and mix evenly. The coefficient of variation (CV) should be ≤5.0%, and the moisture content should be 65%–70%.

[0217] (3) Degradable materials

[0218] Add 0.26 kg of compound bacteria (18.4% termite fungi, 18.4% Trichoderma longipetiae, 26.3% Phanerochaete chrysosporium, 21.1% Aspergillus niger, and 15.8% Bacillus subtilis) to the mixed materials, resulting in an effective viable count of 1.5–2 × 10⁻⁶ bacteria. 8 The mixture should be prepared in batches of 100g each, with a coefficient of variation (CV) ≤ 5.0%. The material should be kept at a moisture content of 40%–60% and the temperature of 37℃–40℃ for the first fermentation, which should last for 7 days.

[0219] (4) High bacterial protein materials

[0220] Take the decomposed material and inoculate it with 0.32 kg of compound mycelium (60% Russula rubra and 40% Codonopsis pilosula mycelium) under positive pressure and aseptic conditions, mixing thoroughly. The coefficient of variation (CV) should be ≤5.0%, and the temperature should be controlled between 20 and 30℃. After inoculation, place it in a sterile room for cultivation, maintaining the cultivation temperature between 22℃ and 30℃ with timely ventilation, humidity between 70% and 85%, and CO2 concentration ≤3.5×10⁻⁶. 3 ppm, culture time 7 days.

[0221] (5) Fermented high microbial protein materials

[0222] Take high mycelial protein material, add 0.37 kg of compound bacteria (44.4% brewer's yeast and 55.5% bifidobacteria), mix evenly, with a coefficient of variation CV ≤ 5.0%, maintain material moisture at 40%–60%, and carry out a second fermentation at 37℃–40℃ for 6 weeks.

[0223] Example 19

[0224] (1) Digestion and sterilization substrate.

[0225] Dandelion (whole plant, 1 kg) and Jerusalem artichoke stalks (20 kg) were washed, dried, crushed, and passed through a 30-mesh sieve to obtain pretreatment material. The pretreatment material was then placed in a high-temperature, high-pressure device for continuous high-temperature, high-pressure steam expansion, explosion, and sterilization (temperature: 180℃, pressure: 10MPa, duration: 30 min, air volume to pretreatment material volume weight ratio: 2L:1kg). After sterilization, the material was cooled to approximately 30℃.

[0226] (2) Mixed materials

[0227] Take all the digestible sterilized base material and add 0.42 kg of compound organic nitrogen source (28.6% yeast powder, 14.2% corn steep liquor powder, 28.6% soybean meal powder, and 28.6% peptone), and mix evenly. The coefficient of variation (CV) should be ≤5.0%, and the moisture content should be 65%–70%.

[0228] (3) Degradable materials

[0229] Add 0.32 kg of compound bacteria (termite fungi 15.8%, Trichoderma longipes 26.3%, Phanerochaete chrysosporium 26.3%, Aspergillus niger 18.4%, Bacillus subtilis 13.2%) to the mixed materials, resulting in an effective viable count of 1.5–2 × 10⁻⁶ bacteria. 8 The mixture should be prepared in batches of 100g each, with a coefficient of variation (CV) ≤ 5.0%. The material should be kept at a moisture content of 40%–60% and the temperature of 37℃–40℃ for the first fermentation, which should last for 7 days.

[0230] (4) High bacterial protein materials

[0231] Take the decomposed material and inoculate it with 0.43 kg of compound mycelium (33.3% Russula rubra and 66.7% Codonopsis pilosula mycelium) under positive pressure and aseptic conditions, mixing thoroughly. The coefficient of variation (CV) should be ≤5.0%, and the temperature should be controlled between 20 and 30℃. After inoculation, place it in a sterile room for cultivation, maintaining the temperature between 22℃ and 30℃ with timely ventilation, humidity between 70% and 85%, and CO2 concentration ≤3.5×10⁻⁶. 3 ppm, culture time 7 days.

[0232] (5) Fermented high microbial protein materials

[0233] Take high mycelial protein material, add 0.44 kg of compound bacteria (58.8% brewer's yeast and 41.2% bifidobacteria), mix evenly, with a coefficient of variation CV ≤ 5.0%, maintain material moisture at 40%–60%, and carry out a second fermentation at 37℃–40℃ for 6 weeks.

[0234] Comparative Example 2

[0235] After washing and drying dandelion (whole plant, 1 kg) and Jerusalem artichoke stalks (20 kg), crush them and pass them through a 30-mesh sieve to obtain the pre-processed material.

[0236] Comparative Example 3

[0237] (1) Digestion and sterilization substrate.

[0238] Dandelion (whole plant, 1 kg) and Jerusalem artichoke stalks (20 kg) were washed, dried, crushed, and passed through a 30-mesh sieve to obtain pretreatment material. The pretreatment material was then placed in a high-temperature, high-pressure device for continuous high-temperature, high-pressure steam expansion, explosion, and sterilization (temperature: 180℃, pressure: 10MPa, duration: 30 min, air volume to pretreatment material volume weight ratio: 2L:1kg). After sterilization, the material was cooled to approximately 30℃.

[0239] (2) Mixed materials

[0240] Take all the digestible sterilized base material and add 0.02 kg (0.1%) of compound organic nitrogen source (28.6% yeast powder, 14.2% corn steep liquor powder, 28.6% soybean meal powder, 28.6% peptone), and mix evenly. The coefficient of variation (CV) should be ≤5.0%, and the moisture content should be 65%–70%.

[0241] (3) Degradable materials

[0242] Add 0.02 kg (0.1%) of compound bacteria (termite fungi 15.8%, Trichoderma longipes 26.3%, Phanerochaete chrysosporium 26.3%, Aspergillus niger 18.4%, Bacillus subtilis 13.2%) to the mixed materials, with an effective viable count of 1.5–2 × 10⁻⁶. 8 The mixture should be prepared in batches of 100g each, with a coefficient of variation (CV) ≤ 5.0%. The material should be kept at a moisture content of 40%–60% and the temperature of 37℃–40℃ for the first fermentation, which should last for 6 days.

[0243] (4) High bacterial protein materials

[0244] Take the decomposed material and inoculate it under positive pressure and aseptic conditions with 0.02 kg (0.1%) of compound mycelium (33.3% Russula rubra and 66.7% Codonopsis pilosula mycelium), mixing thoroughly. The coefficient of variation (CV) should be ≤5.0%, and the temperature should be controlled between 20 and 30℃. After inoculation, place it in a sterile room for cultivation, maintaining the temperature between 22℃ and 30℃ with timely ventilation, and the humidity between 70% and 85%, while keeping the CO2 level low. 2 Concentration ≤ 3.5 × 10³ ppm, incubation time 6 days.

[0245] (5) Fermented high microbial protein materials

[0246] Take high mycelial protein material, add 0.11 kg (0.5%) of compound bacteria (58.8% brewer's yeast and 41.2% bifidobacteria), mix evenly, with a coefficient of variation CV ≤ 5.0%, maintain material moisture at 40%–60%, and carry out a second fermentation at 37℃–40℃ for 5 weeks.

[0247] Comparative Example 4

[0248] (1) Digestion and sterilization substrate.

[0249] Dandelion (whole plant, 1 kg) and Jerusalem artichoke stalks (20 kg) were washed, dried, crushed, and passed through a 30-mesh sieve to obtain pretreatment material. The pretreatment material was then placed in a high-temperature, high-pressure device for continuous high-temperature, high-pressure steam expansion, explosion, and sterilization (temperature: 180℃, pressure: 10MPa, duration: 30 min, air volume to pretreatment material volume weight ratio: 2L:1kg). After sterilization, the material was cooled to approximately 30℃.

[0250] (2) Mixed materials

[0251] Take all the digestible sterilized base material and add 0.53 kg (2.5%) of compound organic nitrogen source (28.6% yeast powder, 14.2% corn steep liquor powder, 28.6% soybean meal powder, 28.6% peptone), and mix evenly. The coefficient of variation (CV) should be ≤5.0%, and the moisture content should be 65%–70%.

[0252] (3) Degradable materials

[0253] Add 0.43 kg (2%) of compound bacteria (termite fungi 15.8%, Trichoderma longipes 26.3%, Phanerochaete chrysosporium 26.3%, Aspergillus niger 18.4%, Bacillus subtilis 13.2%) to the mixed materials, with an effective viable count of 1.5–2 × 10⁻⁶. 8 The mixture should be prepared in batches of 100g each, with a coefficient of variation (CV) ≤ 5.0%. The material should be kept at a moisture content of 40%–60% and the temperature of 37℃–40℃ for the first fermentation, which should last for 6 days.

[0254] (4) High bacterial protein materials

[0255] Take the decomposed material and inoculate it under positive pressure and aseptic conditions with 0.55 kg (2.5%) of compound mycelium (33.3% Russula rubra and 66.7% Codonopsis pilosula mycelium), mixing thoroughly. The coefficient of variation (CV) should be ≤5.0%, and the temperature should be controlled between 20 and 30℃. After inoculation, place it in a sterile room for cultivation, maintaining the temperature between 22℃ and 30℃ with timely ventilation, and the humidity between 70% and 85%, while keeping the CO2 level within a controlled range. 2 Concentration ≤ 3.5 × 10³ ppm, incubation time 6 days.

[0256] (5) Fermented high microbial protein materials

[0257] Take high mycelial protein material, add 0.56 kg (2.5%) of compound bacteria (58.8% brewer's yeast and 41.2% bifidobacteria), mix evenly, with a coefficient of variation CV ≤ 5.0%, maintain material moisture at 40% to 60%, and carry out a second fermentation at 37℃ to 40℃ for 5 weeks.

[0258] Comparative Example 5

[0259] (1) Digestion and sterilization substrate.

[0260] Dandelion (whole plant, 1 kg) and Jerusalem artichoke stalks (20 kg) were washed, dried, crushed, and passed through a 30-mesh sieve to obtain pretreatment material. The pretreatment material was then placed in a high-temperature, high-pressure device for continuous high-temperature, high-pressure steam expansion, explosion, and sterilization (temperature: 180℃, pressure: 10MPa, duration: 30 min, air volume to pretreatment material volume weight ratio: 2L:1kg). After sterilization, the material was cooled to approximately 30℃.

[0261] (2) Mixed materials

[0262] Take all the digestible sterilized base material and add 0.63 kg (3%) of compound organic nitrogen source (28.6% yeast powder, 14.2% corn steep liquor powder, 28.6% soybean meal powder, 28.6% peptone), and mix evenly. The coefficient of variation (CV) should be ≤5.0%, and the moisture content should be 65%–70%.

[0263] (3) Degradable materials

[0264] Add 0.54 kg (2.5%) of a compound microbial agent (termite fungus 15.8%, Trichoderma longipes 26.3%, Phanerochaete chrysosporium 26.3%, Aspergillus niger 18.4%, Bacillus subtilis 13.2%) to the mixed material, resulting in an effective viable count of 1.5–2 × 10⁻⁶. 8 The mixture should be prepared in batches of 100g each, with a coefficient of variation (CV) ≤ 5.0%. The material should be kept at a moisture content of 40%–60% and the temperature of 37℃–40℃ for the first fermentation, which should last for 6 days.

[0265] (4) High bacterial protein materials

[0266] Take the decomposed material and inoculate it under positive pressure and aseptic conditions with 0.67 kg (3%) of compound mycelium (33.3% Russula rubra and 66.7% Codonopsis pilosula mycelium), mixing thoroughly. The coefficient of variation (CV) should be ≤5.0%, and the temperature should be controlled between 20 and 30℃. After inoculation, place it in a sterile room for cultivation, maintaining the cultivation temperature between 22℃ and 30℃ with timely ventilation, and the humidity between 70% and 85%, while keeping the CO2 level within a controlled range. 2 Concentration ≤ 3.5 × 10³ ppm, incubation time 6 days.

[0267] (5) Fermented high microbial protein materials

[0268] Take high mycelial protein material, add 0.69 kg (3%) of compound bacteria (58.8% brewer's yeast and 41.2% bifidobacteria), mix evenly, with a coefficient of variation CV ≤ 5.0%, maintain material moisture at 40% to 60%, and carry out a second fermentation at 37℃ to 40℃ for 5 weeks.

[0269] The high-microbial-protein fermentation materials of Examples 8-19, the pretreatment materials of Comparative Example 2, and the high-microbial-protein fermentation materials of Comparative Examples 3-5 were used to determine the crude protein content, crude fiber content, and phenolic acid content.

[0270] Crude protein content was determined according to the method in GB / T 6432-2018.

[0271] Crude fiber content was determined according to the method in GB / T 6434-2022.

[0272] The phenolic acid content was determined according to the NY / T 3949-2021 method.

[0273] The test results are shown in Table 2. The average crude protein content of dandelion and Jerusalem artichoke straw materials treated by the method of the present invention can reach 20.49%, the crude fiber degradation can reach more than 30%, and the average phenolic acid content can reach 7.46%.

[0274] Table 2. Crude protein, crude fiber, and phenolic acid content of straw materials treated under different embodiment and comparative conditions.

[0275]

[0276]

[0277] Egg-laying hen experiment

[0278] The experiment employed a single-factor, completely randomized experimental design, selecting 500 Hy-Line Brown laying hens at 20 weeks of age with good condition, similar weight, and similar egg production rates. These hens were randomly divided into 5 groups, with 5 replicates per group and 20 hens per replicate (Examples 20-23, Comparative Example 6). All experimental hens had free access to feed and water. The basal diet was formulated according to the nutritional standards of NY / T33-2004 "Chicken Feeding Standards," and the nutritional level of this basal diet was calculated based on the "Chinese Feed Composition and Nutritional Value Table" (2020, 31st edition), as detailed in Table 3. The preliminary experiment lasted 3 days, and the formal experimental period was 64 days.

[0279] Table 3. Composition and nutrient levels of basal diets (dry matter basis)

[0280]

[0281]

[0282] The effects on laying hen production performance, egg quality, immune indicators, and antioxidant capacity were determined.

[0283] Production performance:

[0284] Egg production rate (%) = (Number of eggs produced / Number of chickens) × 100%;

[0285] Average egg weight (g / egg) = Total egg weight / Number of eggs laid;

[0286] Daily feed intake (g / (chickens*d)) = (feed amount - leftover feed) / (number of chickens × number of days in the experiment);

[0287] Feed intake to egg ratio = feed intake / total egg weight.

[0288] Egg freshness and quality:

[0289] Egg shape index = transverse diameter / longitudinal diameter × 100%. Measure the longitudinal diameter and maximum transverse diameter of the egg using vernier calipers, accurate to 0.01 mm.

[0290] Eggshell strength: Measured using a fully automatic eggshell strength tester (ST120H, Shengtai, CN);

[0291] Eggshell thickness: The eggshell thickness at the blunt end, middle and sharp end of the egg is measured using a micrometer, and the average value is calculated to an accuracy of 0.01 mm.

[0292] Yolk color, albumen height, and Haugh units were measured using a multi-functional egg quality analyzer (EA-01, ORKA, USA).

[0293] Immune markers:

[0294] IgA, IgM, and IgG were all measured using an ELISA kit.

[0295] Antioxidant capacity:

[0296] T-AOC, CAT, SOD, GSH, and MDA were all measured using biochemical reagent kits.

[0297] Example 20

[0298] Add 0.5% of the fermented high microbial protein feed additive prepared in Example 15 to the basal diet.

[0299] Example 21

[0300] Add 1.0% of the fermented high microbial protein feed additive prepared in Example 15 to the basal diet.

[0301] Example 22

[0302] Add 1.5% of the fermented high microbial protein feed additive prepared in Example 15 to the basal diet.

[0303] Example 23

[0304] Add 2.0% of the fermented high microbial protein feed additive prepared in Example 15 to the basal diet.

[0305] Comparative Example 6

[0306] Feed only the basic diet.

[0307] The results are shown in Table 4. The fermented high microbial protein feed additive prepared by this invention increased the egg production rate of laying hens by 5.04 percentage points, increased the average egg weight by 5.61%, and enhanced the body's immune function and antioxidant capacity.

[0308] Table 4. Effects of different embodiments and comparative examples on laying hen production performance, egg quality, immune indicators, and antioxidant capacity.

[0309]

[0310] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A method for preparing a fermented straw feed protein additive, characterized in that, Includes the following steps: (1) Dandelion and Jerusalem artichoke stalks are crushed and then steam-exploded to obtain pretreated base material; the mass ratio of dandelion and Jerusalem artichoke stalks is (0.1~1.5):(10~25); (2) After mixing the pretreated substrate described in step (1) with an organic nitrogen source, the first compound bacteria are inoculated to carry out the first fermentation to obtain the degraded material; the temperature of the first fermentation is 37~40℃ and the time of the first fermentation is 5~7 days. The first compound microorganism comprises, by weight, 30-50 parts of termite fungi, 30-50 parts of Trichoderma longipes, 30-50 parts of Phanerochaete chrysosporium, 30-50 parts of Aspergillus niger, and 10-30 parts of Bacillus subtilis; the organic nitrogen source is 0.5-2% of the mass of the pretreated substrate, and the organic nitrogen source includes a compound organic nitrogen source, which, by weight, comprises: 5-10 parts of yeast powder, 5-10 parts of corn steep liquor powder, 5-10 parts of soybean meal powder, and 5-10 parts of peptone; the inoculum size of the first compound microorganism is 0.5-1.5%, and the effective viable count in the bacterial solution of the first compound microorganism is 1.5-2 × 10⁻⁶. 8 pcs / g; (3) After inoculating the degradation material described in step (2) with composite mycelium, a high mycelial protein material is obtained; the composite mycelium includes mycelium of Russula rubra and Codonopsis pilosula; by mass, the composite mycelium includes: 10-30 parts of Russula rubra and 10-30 parts of Codonopsis pilosula; the culture temperature is 22-30℃, the humidity is 70-85%, ventilation is provided as needed, and the CO2 concentration is ≤3.5×10 3 The concentration was ppm, and the culture time was 5-7 days; the mass of the composite mycelium was 0.5-2% of the mass of the pretreated substrate. (4) The high-microbial-protein material described in step (3) is subjected to a second fermentation using the second compound bacteria to obtain a fermented high-microbial-protein material, which is then dried and pulverized to obtain the fermented straw feed protein additive; by mass, the second compound bacteria includes: 30-50 parts of brewer's yeast and 30-50 parts of Bifidobacterium; during the second fermentation, the humidity of the fermentation material is maintained at 40-60%, the temperature at 37-40℃, and the fermentation time is 4-6 weeks; the inoculum amount of the second compound bacteria is 1-2%, and the effective viable count in the bacterial solution of the second compound bacteria is 0.5-1×10 8 per g.

2. The preparation method according to claim 1, characterized in that, The drying method described in step (4) includes the flowing hot air drying method.

3. The preparation method according to claim 2, characterized in that, The parameters of the flowing hot air drying method include: air temperature of 60~70℃ and air flow rate of 1.5~3 m / s.

4. A fermented straw feed protein additive prepared by the preparation method according to any one of claims 1 to 3.

5. The application of the fermented straw feed protein additive according to claim 4 in increasing the crude protein and phenolic acid content in feed, characterized in that, The added mass of the fermented straw feed protein additive is 0.5-2% of the mass of the base feed.

Images