A bacterial enzyme peptide feed additive for preventing pathogenic stiff cattle disease and stiff sheep disease and its preparation method

Abstract

The present invention relates to the field of feed additives, specifically a bacterial enzyme peptide feed additive for preventing pathogenic stunted cow disease and stunted sheep disease, and a preparation method thereof. The present invention uses soybean meal, corn stalks, wheat bran, and rapeseed meal as raw materials, and pre-treats the raw materials by crushing, acid, and alkali to obtain pre-enzyme hydrolysis raw materials; the obtained pre-enzyme hydrolysis raw materials are mixed with coral mucus, and enzymolyzed with a composite enzyme to obtain an enzymatic solution; the enzymatic solution is then inoculated with a composite fermentation bacteria for fermentation to obtain a fermentation liquid; and finally, the fermentation liquid is spray-dried to obtain a bacterial enzyme peptide feed additive for preventing pathogenic stunted cow disease and stunted sheep disease with high stability, good preventive effect against pathogenic stunted cow disease, and low cost.

Description

Technical Field

[0001] The invention relates to the field of feed additives, in particular to a bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease and a preparation method thereof. Background Art

[0002] Stiff cattle and sheep disease is a common disease in cattle and sheep, characterized by symptoms such as slow growth and development, muscle stiffness, and difficulty moving, seriously impacting cattle health and the economic profitability of livestock farming. The disease is associated with a variety of factors, including malnutrition, metabolic disorders, parasitic infections, chronic diseases, and poor husbandry environments. Traditional prevention and treatment methods rely primarily on improving feed nutrition, deworming, and medication, but these methods often have limited effectiveness, high costs, and significant side effects.

[0003] In recent years, with the rapid development of microbiology and enzyme engineering technologies, bacterial enzyme peptide feed additives have gradually attracted attention as a new type of functional feed additive. Combining the multiple functions of probiotics, enzyme preparations, and bioactive peptides, bacterial enzyme peptide feed additives can effectively prevent the occurrence of stiff cow disease and stiff sheep disease by improving digestion and absorption, enhancing immunity, and regulating metabolic function in cattle and sheep. Probiotics can regulate the balance of intestinal flora and inhibit the growth of pathogens; enzyme preparations can break down complex nutrients in feed and improve feed utilization; and bioactive peptides have the effects of enhancing immunity, promoting growth, and providing antioxidant effects.

[0004] However, existing bacterial enzyme peptide feed additives still have some problems in practical applications. First, the product stability is poor: the probiotics, enzyme preparations and active peptides in bacterial enzyme peptide feed additives are easily inactivated during storage and transportation, especially under high temperature, high humidity or long-term storage conditions. Second, the production cost is high: the preparation process of bacterial enzyme peptide feed additives is complex, and the cost of culture medium and raw materials is high, resulting in expensive products. Finally, the specificity and targeting are insufficient: the causes of stiff cow disease and stiff sheep disease are complex, involving multiple factors such as nutrition, metabolism, and environment. Existing additives mostly focus on improving digestion or enhancing immunity, but the targeted effect on pathogenic stiff cow disease and stiff sheep disease is limited. Therefore, how to improve the stability of bacterial enzyme peptide feed for preventing stiff cow disease and stiff sheep disease, reduce costs, and enhance the prevention of pathogenic stiff cow disease has become a major research direction. Summary of the Invention

[0005] In response to the shortcomings of the prior art, the present invention provides a bacterial enzyme peptide feed additive for preventing pathogenic stunted cow disease and stunted sheep disease, and a preparation method thereof. The invention aims to develop a bacterial enzyme peptide feed additive for pathogenic stunted cow disease and stunted sheep disease by optimizing the bacterial strain combination, enzyme preparation formula, and active peptide preparation process, thereby improving its stability, reducing production costs, and enhancing its effectiveness in preventing pathogenic stunted cow disease and stunted sheep disease. By utilizing agricultural by-products as raw materials and combining advanced fermentation and enzymatic hydrolysis technologies, this patent can not only effectively prevent stunted cow disease and stunted sheep disease, but also promote the recycling of resources and promote the development of green farming.

[0006] A method for preparing a bacterial enzyme peptide feed additive for preventing pathogenic stiff cattle disease and stiff sheep disease comprises the following steps:

[0007] (1) Pretreatment: The raw material is crushed into particles with a particle size of 1-2 mm, and the crushed raw material is added to a 0.5-1 wt% hydrochloric acid aqueous solution and acid-treated at a temperature of 90-100 ° C for 1-2 hours; the acid-treated raw material is then washed with water until neutral, and a 1-2 wt% sodium hydroxide aqueous solution is added, and the reaction is carried out at 75-85 ° C for 1-2 hours, washed with water until neutral, and dried to obtain a pre-enzymatic hydrolysis raw material;

[0008] (2) adding the pre-enzymatic hydrolysis raw material and coral mucus obtained in step (1) to the compound enzyme hydrolysis solution, performing enzymatic hydrolysis at 50-60° C. for 4-6 hours, filtering to remove solid residue, and obtaining an enzymatic hydrolysis solution;

[0009] (3) inoculating the composite fermentation bacteria into the enzymatic hydrolysate obtained in step (2) at an inoculation amount of 5-10 wt %, and fermenting at 30-37° C. and a pH of 4.5-5.5 for 24-48 hours. During the fermentation process, stirring was performed at a speed of 800-1000 r / min, and oxygen was supplied to maintain the dissolved oxygen content at 60-80%. After the fermentation was completed, centrifugation was performed at a speed of 800-1200 r / min for 4-6 minutes to obtain a fermentation liquid;

[0010] (4) The fermentation liquid obtained in step (3) is spray-dried at a temperature of 60-80° C. to obtain the bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease.

[0011] The raw material in step (1) is at least one of soybean meal, corn straw, wheat bran, rice bran and rapeseed meal; preferably, the raw material is a mixture of soybean meal, corn straw, wheat bran and rapeseed meal in a mass ratio of (1-2): (3-4): (2-3): (1-2).

[0012] In step (1), the mass ratio of the hydrochloric acid aqueous solution to the raw material is (1-10):1, and the mass ratio of the sodium hydroxide aqueous solution to the raw material is (1-10):1.

[0013] The mass ratio of the pre-enzymatic hydrolysis raw material, coral mucus, and compound enzyme hydrolyzate in step (2) is 1: (0.1-0.4): (1-2).

[0014] The enzyme in the compound enzyme hydrolysis solution of step (2) is at least one of trypsin, amylase, and cellulase; preferably, the enzyme in the compound enzyme hydrolysis solution is a mixture of trypsin, amylase, and cellulase, wherein the concentration of trypsin is 0.5-1wt%, the concentration of amylase is 0.5-1wt%, the concentration of cellulase is 1-2wt%, and the balance is water.

[0015] The composite fermentation bacteria in step (3) is at least one of thermophilic Streptococcus, Aspergillus niger, and Bacillus subtilis; preferably, the composite fermentation bacteria is a mixture of thermophilic Streptococcus, Aspergillus niger, and Bacillus subtilis in a mass ratio of (1-2): (1-2): 2.

[0016] Soybean meal is rich in plant protein and can be used to produce bioactive peptides through enzymatic hydrolysis or fermentation.

[0017] Corn stover is rich in cellulose and can serve as a fermentation substrate for probiotics.

[0018] Wheat bran is rich in dietary fiber and a small amount of protein, and can be used for probiotic culture or enzymatic hydrolysis to prepare functional peptides.

[0019] Rapeseed meal is rich in protein and is an important source of peptides.

[0020] The present invention uses soybean meal, corn straw, wheat bran and rapeseed meal as raw materials, mixes various agricultural by-products, optimizes the nutritional components of the fermentation substrate, and at the same time promotes the recycling of resources and promotes the development of green agriculture.

[0021] The invention adopts crushing, acid and alkali to pretreat raw materials to obtain pre-enzyme hydrolysis raw materials; the obtained pre-enzyme hydrolysis raw materials and coral mucus are mixed, and enzymolysis is performed with a composite enzyme to obtain an enzymatic solution; composite fermentation bacteria are then inoculated into the enzymatic solution for fermentation to obtain a fermentation liquid; and finally the fermentation liquid is spray-dried to obtain the bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease.

[0022] First, acid-base cooking treatment can remove some anti-nutritional factors (such as natural inhibitors and antioxidants in plants), destroy plant cell walls, release more soluble proteins and polysaccharides, provide better substrates for subsequent enzymatic hydrolysis, and provide a suitable environment for subsequent fermentation.

[0023] The use of enzymes with broad-spectrum enzymatic hydrolysis capabilities (such as pepsin, papain, trypsin, etc.) can degrade large molecular proteins in the raw materials into small peptides or amino acids. Small peptides have certain antibacterial or immunomodulatory effects, enhance the animal's disease resistance, and indirectly reduce the risk of pathogen infection.

[0024] Because cattle's staple diet contains a large amount of plant materials, the cellulose and hemicellulose in these materials provide a suitable environment for pathogens (such as E. coli, Salmonella, and Brucella). During the enzymatic hydrolysis process, the use of cellulase effectively degrades plant fibers, increasing the release of nutrients. It also alters the intestinal environment, indirectly affecting the growth of pathogens.

[0025] Amylase can degrade starch in plants, releasing monosaccharides or oligosaccharides, which promotes nutrient utilization. The small molecular sugars produced during starch degradation can also provide substrates for fermentation, promoting the growth and reproduction of beneficial bacteria.

[0026] According to the different target raw materials, appropriate enzyme composition types are selected and the enzymatic hydrolysis formula is optimized, which can improve the fermentation efficiency and further enhance the specificity of the fermentation product to pathogenic rickets.

[0027] Streptococcus thermophilus has broad-spectrum antimicrobial properties and can lower the pH of the environment by producing lactic acid, inhibiting the growth of anaerobic pathogens. Furthermore, Streptococcus thermophilus can promote intestinal health and strengthen the immune system, thereby improving disease resistance.

[0028] Aspergillus niger secretes a variety of enzymes, particularly cellulases, amylases, and proteases. These enzymes excel at digesting and breaking down complex organic matter (such as plant residues), thereby improving feed digestibility and utilization. Furthermore, Aspergillus niger secretes a wider variety of enzymes during the fermentation process, helping to break down cellulose, starch, and protein in plant-derived feedstocks (such as soybean meal, corn stover, and rapeseed meal), releasing more nutrients, improving feed quality, and indirectly preventing cow and sheep stiffness.

[0029] Bacillus subtilis is widely used in feed and exhibits antimicrobial activity. It can inhibit the growth of anaerobic bacteria by secreting antimicrobial peptides. By optimizing fermentation conditions, the activity of Bacillus subtilis during fermentation can be increased, enhancing its anti-disease effects.

[0030] By using thermophilic Streptococcus, Aspergillus niger and Bacillus together, a complementary effect can be achieved, and the beneficial substances produced by each of them (such as lactic acid, antimicrobial peptides, ethanol, etc.) can be used to synergistically inhibit pathogens. They work synergistically during the fermentation process to produce more diverse active substances and enhance the functional diversity of bacterial enzyme peptides.

[0031] Coral mucus also contains some antioxidants, which can help resist oxidation and protect the activity of enzymes and peptides during the spray drying process.

[0032] As a natural source of polysaccharides and proteins, coral mucus may enhance the stability of zymopeptides, particularly by protecting the membrane during the spray-drying process and preventing the loss of enzyme and peptide activity. Furthermore, coral mucus may also improve the yield and quality of zymopeptides by providing nutrients and promoting microbial growth during enzymatic hydrolysis and fermentation.

[0033] Adding coral mucus to the enzymatic hydrolysis step, as it contains polysaccharides and proteins, may provide an additional sugar source for the enzymatic hydrolysis reaction, helping to improve enzyme efficiency. The proteins and sugars in the coral mucus can also promote the growth of bacteria such as thermophilic Streptococcus, Aspergillus niger, and Bacillus, thereby increasing the yield of enzymatic peptides. Furthermore, the antioxidants in the coral mucus may help maintain the stability of the fermentation system and reduce the negative effects of environmental oxidation on microorganisms and peptides. The sugars in the coral mucus may form a protective film during the spray drying process, preventing excessive evaporation of water and reducing the inactivation of enzymes and peptides during drying. Furthermore, the proteins in the mucus may react with the enzyme peptides to form stable complexes, thereby preventing denaturation or degradation of the enzyme peptides.

[0034] Beneficial effects of the present invention:

[0035] The invention uses soybean meal, corn stalks, wheat bran and rapeseed meal as raw materials, performs pretreatment by crushing, acid and alkali, performs enzymolysis by composite enzyme, ferments by composite fermentation bacteria and spray-dries to obtain the bacterial enzyme peptide feed additive for preventing pathogenic scurvy and scurvy. Coral mucus is added for synergistic enzymolysis and fermentation to obtain a bacterial enzyme peptide feed additive for preventing pathogenic scurvy and scurvy, which has high stability, good preventive effect on pathogenic scurvy and scurvy and low cost. DETAILED DESCRIPTION

[0036] Soybean meal, corn straw, wheat bran, and rapeseed meal are conventional agricultural by-products and are used as raw materials after disinfection and sterilization.

[0037] Streptococcus thermophilus, accession number: CGMCC 1.1855, was purchased from China General Microbiological Culture Collection Center.

[0038] Aspergillus niger, collection number: CICC 2487, was purchased from China Industrial Microorganism Culture Collection Administration Center.

[0039] Bacillus subtilis, accession number: CGMCC 1.1086, was purchased from China General Microbiological Culture Collection Center.

[0040] Salmonella enterica subsp. enterica, strain number: CGMCC 1.1859, was purchased from China General Microbiological Culture Collection Center.

[0041] Trypsin, product number: 27250018, Thermo Fisher Scientific (China) Co., Ltd.

[0042] Amylase, product number: S68733, Shanghai Yuanye Biotechnology Co., Ltd.

[0043] Cellulase, product number: S10041, Shanghai Yuanye Biotechnology Co., Ltd.

[0044] Coral mucus: Giant synphyllous corals from the waters of Sanya, Hainan were used. The living corals were stimulated by striking and then inverted to collect the mucus. The obtained mucus and activated carbon particles (Article No.: YK-10-18, Pingdingshan Lvlin Activated Carbon Co., Ltd.) were mixed and stirred in a mass ratio of 1:0.1, and filtered to obtain the required coral mucus.

[0045] Example 1

[0046] A method for preparing a bacterial enzyme peptide feed additive for preventing pathogenic stiff cattle disease and stiff sheep disease comprises the following steps:

[0047] (1) Pretreatment: The raw material was crushed into particles with a particle size of 1 mm, and the crushed raw material was added to a 0.6 wt% hydrochloric acid aqueous solution and acid-treated at a temperature of 95 ° C for 1.5 hours; then the acid-treated raw material was washed with water until neutral, and a 1.5 wt% sodium hydroxide aqueous solution was added and treated at 80 ° C for 1.5 hours, washed with water until neutral, and dried to obtain a pre-enzymatic hydrolysis raw material;

[0048] (2) adding the pre-enzymatic hydrolysis raw material and coral mucus obtained in step (1) to the compound enzyme hydrolysis solution, performing enzymatic hydrolysis at 55° C. for 5 hours, filtering to remove solid residue, and obtaining an enzymatic hydrolysis solution;

[0049] (3) inoculating the composite fermentation bacteria into the enzymatic hydrolysate obtained in step (2) at an inoculation amount of 7 wt %, and fermenting at 34° C. and a pH of 5 for 36 hours. During the fermentation process, stirring was performed at a speed of 800 r / min, and oxygen was supplied to maintain the dissolved oxygen content at 70%. After the fermentation was completed, centrifugation was performed at a speed of 1000 r / min for 5 minutes to obtain a fermentation liquid;

[0050] (4) The fermentation liquid obtained in step (3) is spray-dried at a temperature of 70° C. to obtain the bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease.

[0051] The raw materials in step (1) are prepared by mixing soybean meal, corn straw, wheat bran and rapeseed meal in a mass ratio of 1:3:2:1.

[0052] In step (1), the mass ratio of the hydrochloric acid aqueous solution to the raw material is 2:1, and the mass ratio of the sodium hydroxide aqueous solution to the raw material is 2:1.

[0053] The mass ratio of the pre-enzymatic hydrolysis raw material, coral mucus, and compound enzyme hydrolyzate in step (2) is 1:0.3:2.

[0054] The enzymes in the complex enzyme hydrolysis solution of step (2) are a mixture of trypsin, amylase, and cellulase, wherein the concentration of trypsin is 0.6wt%, the concentration of amylase is 0.6wt%, the concentration of cellulase is 1.2wt%, and the balance is water.

[0055] The composite fermentation bacteria in step (3) is a mixture of thermophilic Streptococcus, Aspergillus niger and Bacillus subtilis in a mass ratio of 1:1:2.

[0056] Example 2

[0057] A method for preparing a bacterial enzyme peptide feed additive for preventing pathogenic stiff cattle disease and stiff sheep disease comprises the following steps:

[0058] (1) Pretreatment: The raw material was crushed into particles with a particle size of 1 mm, and the crushed raw material was added to a 0.6 wt% hydrochloric acid aqueous solution and acid-treated at a temperature of 95 ° C for 1.5 hours; then the acid-treated raw material was washed with water until neutral, and a 1.5 wt% sodium hydroxide aqueous solution was added and treated at 80 ° C for 1.5 hours, washed with water until neutral, and dried to obtain a pre-enzymatic hydrolysis raw material;

[0059] (2) adding the pre-enzymatic hydrolysis raw material and coral mucus obtained in step (1) to the compound enzyme hydrolysis solution, performing enzymatic hydrolysis at 55° C. for 5 hours, filtering to remove solid residue, and obtaining an enzymatic hydrolysis solution;

[0060] (3) inoculating the composite fermentation bacteria into the enzymatic hydrolysate obtained in step (2) at an inoculation amount of 7 wt %, and fermenting at 34° C. and a pH of 5 for 36 hours. During the fermentation process, stirring was performed at a speed of 800 r / min, and oxygen was supplied to maintain the dissolved oxygen content at 70%. After the fermentation was completed, centrifugation was performed at a speed of 1000 r / min for 5 minutes to obtain a fermentation liquid;

[0061] (4) The fermentation liquid obtained in step (3) is spray-dried at a temperature of 70° C. to obtain the bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease.

[0062] The raw materials in step (1) are prepared by mixing soybean meal, corn straw, wheat bran and rapeseed meal in a mass ratio of 1:3:2:1.

[0063] In step (1), the mass ratio of the hydrochloric acid aqueous solution to the raw material is 2:1, and the mass ratio of the sodium hydroxide aqueous solution to the raw material is 2:1.

[0064] The mass ratio of the pre-enzymatic hydrolysis raw material, coral mucus, and compound enzyme hydrolyzate in step (2) is 1:0.3:2.

[0065] The enzymes in the complex enzyme hydrolysis solution of step (2) are a mixture of amylase and cellulase, wherein the concentration of amylase is 0.6wt%, the concentration of cellulase is 1.2wt%, and the balance is water.

[0066] The composite fermentation bacteria in step (3) is a mixture of thermophilic Streptococcus, Aspergillus niger and Bacillus subtilis in a mass ratio of 1:1:2.

[0067] Example 3

[0068] A method for preparing a bacterial enzyme peptide feed additive for preventing pathogenic stiff cattle disease and stiff sheep disease comprises the following steps:

[0069] (1) Pretreatment: The raw material was crushed into particles with a particle size of 1 mm, and the crushed raw material was added to a 0.6 wt% hydrochloric acid aqueous solution and acid-treated at a temperature of 95 ° C for 1.5 hours; then the acid-treated raw material was washed with water until neutral, and a 1.5 wt% sodium hydroxide aqueous solution was added and treated at 80 ° C for 1.5 hours, washed with water until neutral, and dried to obtain a pre-enzymatic hydrolysis raw material;

[0070] (2) adding the pre-enzymatic hydrolysis raw material and coral mucus obtained in step (1) to the compound enzyme hydrolysis solution, performing enzymatic hydrolysis at 55° C. for 5 hours, filtering to remove solid residue, and obtaining an enzymatic hydrolysis solution;

[0071] (3) inoculating the composite fermentation bacteria into the enzymatic hydrolysate obtained in step (2) at an inoculation amount of 7 wt %, and fermenting at 34° C. and a pH of 5 for 36 hours. During the fermentation process, stirring was performed at a speed of 800 r / min, and oxygen was supplied to maintain the dissolved oxygen content at 70%. After the fermentation was completed, centrifugation was performed at a speed of 1000 r / min for 5 minutes to obtain a fermentation liquid;

[0072] (4) The fermentation liquid obtained in step (3) is spray-dried at a temperature of 70° C. to obtain the bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease.

[0073] The raw materials in step (1) are prepared by mixing soybean meal, corn straw, wheat bran and rapeseed meal in a mass ratio of 1:3:2:1.

[0074] In step (1), the mass ratio of the hydrochloric acid aqueous solution to the raw material is 2:1, and the mass ratio of the sodium hydroxide aqueous solution to the raw material is 2:1.

[0075] The mass ratio of the pre-enzymatic hydrolysis raw material, coral mucus, and compound enzyme hydrolyzate in step (2) is 1:0.3:2.

[0076] The enzymes in the complex enzyme hydrolysis solution of step (2) are a mixture of trypsin and cellulase, wherein the concentration of trypsin is 0.6 wt %, the concentration of cellulase is 1.2 wt %, and the balance is water.

[0077] The composite fermentation bacteria in step (3) is a mixture of thermophilic Streptococcus, Aspergillus niger and Bacillus subtilis in a mass ratio of 1:1:2.

[0078] Example 4

[0079] A method for preparing a bacterial enzyme peptide feed additive for preventing pathogenic stiff cattle disease and stiff sheep disease comprises the following steps:

[0080] (1) Pretreatment: The raw material was crushed into particles with a particle size of 1 mm, and the crushed raw material was added to a 0.6 wt% hydrochloric acid aqueous solution and acid-treated at a temperature of 95 ° C for 1.5 hours; then the acid-treated raw material was washed with water until neutral, and a 1.5 wt% sodium hydroxide aqueous solution was added and treated at 80 ° C for 1.5 hours, washed with water until neutral, and dried to obtain a pre-enzymatic hydrolysis raw material;

[0081] (2) adding the pre-enzymatic hydrolysis raw material and coral mucus obtained in step (1) to the compound enzyme hydrolysis solution, performing enzymatic hydrolysis at 55° C. for 5 hours, filtering to remove solid residue, and obtaining an enzymatic hydrolysis solution;

[0082] (3) inoculating the composite fermentation bacteria into the enzymatic hydrolysate obtained in step (2) at an inoculation amount of 7 wt %, and fermenting at 34° C. and a pH of 5 for 36 hours. During the fermentation process, stirring was performed at a speed of 800 r / min, and oxygen was supplied to maintain the dissolved oxygen content at 70%. After the fermentation was completed, centrifugation was performed at a speed of 1000 r / min for 5 minutes to obtain a fermentation liquid;

[0083] (4) The fermentation liquid obtained in step (3) is spray-dried at a temperature of 70° C. to obtain the bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease.

[0084] The raw materials in step (1) are prepared by mixing soybean meal, corn straw, wheat bran and rapeseed meal in a mass ratio of 1:3:2:1.

[0085] In step (1), the mass ratio of the hydrochloric acid aqueous solution to the raw material is 2:1, and the mass ratio of the sodium hydroxide aqueous solution to the raw material is 2:1.

[0086] The mass ratio of the pre-enzymatic hydrolysis raw material, coral mucus, and compound enzyme hydrolyzate in step (2) is 1:0.3:2.

[0087] The enzymes in the complex enzyme hydrolysis solution of step (2) are a mixture of trypsin and amylase, wherein the concentration of trypsin is 0.6 wt %, the concentration of amylase is 0.6 wt %, and the balance is water.

[0088] The composite fermentation bacteria in step (3) is a mixture of thermophilic Streptococcus, Aspergillus niger and Bacillus subtilis in a mass ratio of 1:1:2.

[0089] Example 5

[0090] A method for preparing a bacterial enzyme peptide feed additive for preventing pathogenic stiff cattle disease and stiff sheep disease comprises the following steps:

[0091] (1) Pretreatment: The raw material was crushed into particles with a particle size of 1 mm, and the crushed raw material was added to a 0.6 wt% hydrochloric acid aqueous solution and acid-treated at a temperature of 95 ° C for 1.5 hours; then the acid-treated raw material was washed with water until neutral, and a 1.5 wt% sodium hydroxide aqueous solution was added and treated at 80 ° C for 1.5 hours, washed with water until neutral, and dried to obtain a pre-enzymatic hydrolysis raw material;

[0092] (2) adding the pre-enzymatic hydrolysis raw material and coral mucus obtained in step (1) to the compound enzyme hydrolysis solution, performing enzymatic hydrolysis at 55° C. for 5 hours, filtering to remove solid residue, and obtaining an enzymatic hydrolysis solution;

[0093] (3) inoculating the composite fermentation bacteria into the enzymatic hydrolysate obtained in step (2) at an inoculation amount of 7 wt %, and fermenting at 34° C. and a pH of 5 for 36 hours. During the fermentation process, stirring was performed at a speed of 800 r / min, and oxygen was supplied to maintain the dissolved oxygen content at 70%. After the fermentation was completed, centrifugation was performed at a speed of 1000 r / min for 5 minutes to obtain a fermentation liquid;

[0094] (4) The fermentation liquid obtained in step (3) is spray-dried at a temperature of 70° C. to obtain the bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease.

[0095] The raw materials in step (1) are prepared by mixing soybean meal, corn straw, wheat bran and rapeseed meal in a mass ratio of 1:3:2:1.

[0096] In step (1), the mass ratio of the hydrochloric acid aqueous solution to the raw material is 2:1, and the mass ratio of the sodium hydroxide aqueous solution to the raw material is 2:1.

[0097] The mass ratio of the pre-enzymatic hydrolysis raw material, coral mucus, and compound enzyme hydrolyzate in step (2) is 1:0.3:2.

[0098] The enzymes in the complex enzyme hydrolysis solution of step (2) are a mixture of trypsin, amylase, and cellulase, wherein the concentration of trypsin is 0.6wt%, the concentration of amylase is 0.6wt%, the concentration of cellulase is 1.2wt%, and the balance is water.

[0099] The composite fermentation bacteria in step (3) is prepared by mixing thermophilic Streptococcus and Aspergillus niger in a mass ratio of 1:1.

[0100] Example 6

[0101] A method for preparing a bacterial enzyme peptide feed additive for preventing pathogenic stiff cattle disease and stiff sheep disease comprises the following steps:

[0102] (1) Pretreatment: The raw material was crushed into particles with a particle size of 1 mm, and the crushed raw material was added to a 0.6 wt% hydrochloric acid aqueous solution and acid-treated at a temperature of 95 ° C for 1.5 hours; then the acid-treated raw material was washed with water until neutral, and a 1.5 wt% sodium hydroxide aqueous solution was added and treated at 80 ° C for 1.5 hours, washed with water until neutral, and dried to obtain a pre-enzymatic hydrolysis raw material;

[0103] (2) adding the pre-enzymatic hydrolysis raw material and coral mucus obtained in step (1) to the compound enzyme hydrolysis solution, performing enzymatic hydrolysis at 55° C. for 5 hours, filtering to remove solid residue, and obtaining an enzymatic hydrolysis solution;

[0104] (3) inoculating the composite fermentation bacteria into the enzymatic hydrolysate obtained in step (2) at an inoculation amount of 7 wt %, and fermenting at 34° C. and a pH of 5 for 36 hours. During the fermentation process, stirring was performed at a speed of 800 r / min, and oxygen was supplied to maintain the dissolved oxygen content at 70%. After the fermentation was completed, centrifugation was performed at a speed of 1000 r / min for 5 minutes to obtain a fermentation liquid;

[0105] (4) The fermentation liquid obtained in step (3) is spray-dried at a temperature of 70° C. to obtain the bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease.

[0106] The raw materials in step (1) are prepared by mixing soybean meal, corn straw, wheat bran and rapeseed meal in a mass ratio of 1:3:2:1.

[0107] In step (1), the mass ratio of the hydrochloric acid aqueous solution to the raw material is 2:1, and the mass ratio of the sodium hydroxide aqueous solution to the raw material is 2:1.

[0108] The mass ratio of the pre-enzymatic hydrolysis raw material, coral mucus, and compound enzyme hydrolyzate in step (2) is 1:0.3:2.

[0109] The enzymes in the complex enzyme hydrolysis solution of step (2) are a mixture of trypsin, amylase, and cellulase, wherein the concentration of trypsin is 0.6wt%, the concentration of amylase is 0.6wt%, the concentration of cellulase is 1.2wt%, and the balance is water.

[0110] The composite fermentation bacteria in step (3) is a mixture of thermophilic Streptococcus and Bacillus subtilis in a mass ratio of 1:2.

[0111] Example 7

[0112] A method for preparing a bacterial enzyme peptide feed additive for preventing pathogenic stiff cattle disease and stiff sheep disease comprises the following steps:

[0113] (1) Pretreatment: The raw material was crushed into particles with a particle size of 1 mm, and the crushed raw material was added to a 0.6 wt% hydrochloric acid aqueous solution and acid-treated at a temperature of 95 ° C for 1.5 hours; then the acid-treated raw material was washed with water until neutral, and a 1.5 wt% sodium hydroxide aqueous solution was added and treated at 80 ° C for 1.5 hours, washed with water until neutral, and dried to obtain a pre-enzymatic hydrolysis raw material;

[0114] (2) adding the pre-enzymatic hydrolysis raw material and coral mucus obtained in step (1) to the compound enzyme hydrolysis solution, performing enzymatic hydrolysis at 55° C. for 5 hours, filtering to remove solid residue, and obtaining an enzymatic hydrolysis solution;

[0115] (3) inoculating the composite fermentation bacteria into the enzymatic hydrolysate obtained in step (2) at an inoculation amount of 7 wt %, and fermenting at 34° C. and a pH of 5 for 36 hours. During the fermentation process, stirring was performed at a speed of 800 r / min, and oxygen was supplied to maintain the dissolved oxygen content at 70%. After the fermentation was completed, centrifugation was performed at a speed of 1000 r / min for 5 minutes to obtain a fermentation liquid;

[0116] (4) The fermentation liquid obtained in step (3) is spray-dried at a temperature of 70° C. to obtain the bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease.

[0117] The raw materials in step (1) are prepared by mixing soybean meal, corn straw, wheat bran and rapeseed meal in a mass ratio of 1:3:2:1.

[0118] In step (1), the mass ratio of the hydrochloric acid aqueous solution to the raw material is 2:1, and the mass ratio of the sodium hydroxide aqueous solution to the raw material is 2:1.

[0119] The mass ratio of the pre-enzymatic hydrolysis raw material, coral mucus, and compound enzyme hydrolyzate in step (2) is 1:0.3:2.

[0120] The enzymes in the complex enzyme hydrolysis solution of step (2) are a mixture of trypsin, amylase, and cellulase, wherein the concentration of trypsin is 0.6wt%, the concentration of amylase is 0.6wt%, the concentration of cellulase is 1.2wt%, and the balance is water.

[0121] The composite fermentation bacteria in step (3) is prepared by mixing Aspergillus niger and Bacillus subtilis in a mass ratio of 1:2.

[0122] Example 8

[0123] A method for preparing a bacterial enzyme peptide feed additive for preventing pathogenic stiff cattle disease and stiff sheep disease comprises the following steps:

[0124] (1) Pretreatment: The raw material was crushed into particles with a particle size of 1 mm, and the crushed raw material was added to a 0.6 wt% hydrochloric acid aqueous solution and acid-treated at a temperature of 95 ° C for 1.5 hours; then the acid-treated raw material was washed with water until neutral, and a 1.5 wt% sodium hydroxide aqueous solution was added and treated at 80 ° C for 1.5 hours, washed with water until neutral, and dried to obtain a pre-enzymatic hydrolysis raw material;

[0125] (2) adding the pre-enzymatic hydrolysis raw material obtained in step (1) to the complex enzyme hydrolysis solution, performing enzymatic hydrolysis at 55°C for 5 hours, filtering to remove solid residues, and obtaining an enzymatic hydrolysis solution;

[0126] (3) inoculating the composite fermentation bacteria into the enzymatic hydrolysate obtained in step (2) at an inoculation amount of 7 wt %, and adding coral mucus, and fermenting at 34° C. and pH 5 for 36 hours. During the fermentation process, stirring was performed at a speed of 800 r / min, and oxygen was supplied to maintain the dissolved oxygen content at 70%. After the fermentation was completed, centrifugation was performed at a speed of 1000 r / min for 5 minutes to obtain a fermentation liquid;

[0127] (4) The fermentation liquid obtained in step (3) is spray-dried at a temperature of 70° C. to obtain the bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease.

[0128] The raw materials in step (1) are prepared by mixing soybean meal, corn straw, wheat bran and rapeseed meal in a mass ratio of 1:3:2:1.

[0129] In step (1), the mass ratio of the hydrochloric acid aqueous solution to the raw material is 2:1, and the mass ratio of the sodium hydroxide aqueous solution to the raw material is 2:1.

[0130] The mass ratio of the pre-enzymatic hydrolysis raw material to the complex enzyme hydrolysis solution in step (2) is 1:2.

[0131] The enzymes in the complex enzyme hydrolysis solution of step (2) are a mixture of trypsin, amylase, and cellulase, wherein the concentration of trypsin is 0.6wt%, the concentration of amylase is 0.6wt%, the concentration of cellulase is 1.2wt%, and the balance is water.

[0132] The composite fermentation bacteria in step (3) is a mixture of thermophilic Streptococcus, Aspergillus niger and Bacillus subtilis in a mass ratio of 1:1:2.

[0133] The mass ratio of the coral mucus in step (3) to the pre-enzymatic hydrolysis raw material in step (2) is 0.3:1.

[0134] Example 9

[0135] A method for preparing a bacterial enzyme peptide feed additive for preventing pathogenic stiff cattle disease and stiff sheep disease comprises the following steps:

[0136] (1) Pretreatment: The raw material was crushed into particles with a particle size of 1 mm, and the crushed raw material was added to a 0.6 wt% hydrochloric acid aqueous solution and acid-treated at a temperature of 95 ° C for 1.5 hours; then the acid-treated raw material was washed with water until neutral, and a 1.5 wt% sodium hydroxide aqueous solution was added and treated at 80 ° C for 1.5 hours, washed with water until neutral, and dried to obtain a pre-enzymatic hydrolysis raw material;

[0137] (2) adding the pre-enzymatic hydrolysis raw material obtained in step (1) to the complex enzyme hydrolysis solution, performing enzymatic hydrolysis at 55°C for 5 hours, filtering to remove solid residues, and obtaining an enzymatic hydrolysis solution;

[0138] (3) inoculating the composite fermentation bacteria into the enzymatic hydrolysate obtained in step (2) at an inoculation amount of 7 wt %, and fermenting at 34° C. and a pH of 5 for 36 hours. During the fermentation process, stirring was performed at a speed of 800 r / min, and oxygen was supplied to maintain the dissolved oxygen content at 70%. After the fermentation was completed, centrifugation was performed at a speed of 1000 r / min for 5 minutes to obtain a fermentation liquid;

[0139] (4) Adding coral mucus to the fermentation liquid obtained in step (3), stirring evenly and then spray drying at a temperature of 70° C. to obtain the bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease.

[0140] The raw materials in step (1) are prepared by mixing soybean meal, corn straw, wheat bran and rapeseed meal in a mass ratio of 1:3:2:1.

[0141] In step (1), the mass ratio of the hydrochloric acid aqueous solution to the raw material is 2:1, and the mass ratio of the sodium hydroxide aqueous solution to the raw material is 2:1.

[0142] The mass ratio of the pre-enzymatic hydrolysis raw material to the complex enzyme hydrolysis solution in step (2) is 1:2.

[0143] The enzymes in the complex enzyme hydrolysis solution of step (2) are a mixture of trypsin, amylase, and cellulase, wherein the concentration of trypsin is 0.6wt%, the concentration of amylase is 0.6wt%, the concentration of cellulase is 1.2wt%, and the balance is water.

[0144] The composite fermentation bacteria in step (3) is a mixture of thermophilic Streptococcus, Aspergillus niger and Bacillus subtilis in a mass ratio of 1:1:2.

[0145] The mass ratio of the coral mucus in step (4) to the pre-enzymatic hydrolysis raw material in step (2) is 0.3:1.

[0146] Comparative Example 1

[0147] A method for preparing a bacterial enzyme peptide feed additive for preventing pathogenic stiff cattle disease and stiff sheep disease comprises the following steps:

[0148] (1) Pretreatment: The raw material is crushed into granules with a particle size of 1 mm and dried to obtain the pre-enzymatic hydrolysis raw material;

[0149] (2) adding the pre-enzymatic hydrolysis raw material and coral mucus obtained in step (1) to the compound enzyme hydrolysis solution, performing enzymatic hydrolysis at 55° C. for 5 hours, filtering to remove solid residue, and obtaining an enzymatic hydrolysis solution;

[0150] (3) inoculating the composite fermentation bacteria into the enzymatic hydrolysate obtained in step (2) at an inoculation amount of 7 wt %, and fermenting at 34° C. and a pH of 5 for 36 hours. During the fermentation process, stirring was performed at a speed of 800 r / min, and oxygen was supplied to maintain the dissolved oxygen content at 70%. After the fermentation was completed, centrifugation was performed at a speed of 1000 r / min for 5 minutes to obtain a fermentation liquid;

[0151] (4) The fermentation liquid obtained in step (3) is spray-dried at a temperature of 70° C. to obtain the bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease.

[0152] The raw materials in step (1) are prepared by mixing soybean meal, corn straw, wheat bran and rapeseed meal in a mass ratio of 1:3:2:1.

[0153] The mass ratio of the pre-enzymatic hydrolysis raw material, coral mucus, and compound enzyme hydrolyzate in step (2) is 1:0.3:2.

[0154] The enzymes in the complex enzyme hydrolysis solution of step (2) are a mixture of trypsin, amylase, and cellulase, wherein the concentration of trypsin is 0.6wt%, the concentration of amylase is 0.6wt%, the concentration of cellulase is 1.2wt%, and the balance is water.

[0155] The composite fermentation bacteria in step (3) is a mixture of thermophilic Streptococcus, Aspergillus niger and Bacillus subtilis in a mass ratio of 1:1:2.

[0156] Comparative Example 2

[0157] A method for preparing a bacterial enzyme peptide feed additive for preventing pathogenic stiff cattle disease and stiff sheep disease comprises the following steps:

[0158] (1) Pretreatment: The raw material was crushed into particles with a particle size of 1 mm, and the crushed raw material was added to a 0.6 wt% hydrochloric acid aqueous solution and acid-treated at a temperature of 95 ° C for 1.5 hours; then the acid-treated raw material was washed with water until neutral, and a 1.5 wt% sodium hydroxide aqueous solution was added and treated at 80 ° C for 1.5 hours, washed with water until neutral, and dried to obtain a pre-enzymatic hydrolysis raw material;

[0159] (2) adding the pre-enzymatic hydrolysis raw material obtained in step (1) to the complex enzyme hydrolysis solution, performing enzymatic hydrolysis at 55°C for 5 hours, filtering to remove solid residues, and obtaining an enzymatic hydrolysis solution;

[0160] (3) inoculating the composite fermentation bacteria into the enzymatic hydrolysate obtained in step (2) at an inoculation amount of 7 wt %, and fermenting at 34° C. and a pH of 5 for 36 hours. During the fermentation process, stirring was performed at a speed of 800 r / min, and oxygen was supplied to maintain the dissolved oxygen content at 70%. After the fermentation was completed, centrifugation was performed at a speed of 1000 r / min for 5 minutes to obtain a fermentation liquid;

[0161] (4) The fermentation liquid obtained in step (3) is spray-dried at a temperature of 70° C. to obtain the bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease.

[0162] The raw materials in step (1) are prepared by mixing soybean meal, corn straw, wheat bran and rapeseed meal in a mass ratio of 1:3:2:1.

[0163] In step (1), the mass ratio of the hydrochloric acid aqueous solution to the raw material is 2:1, and the mass ratio of the sodium hydroxide aqueous solution to the raw material is 2:1.

[0164] The mass ratio of the pre-enzymatic hydrolysis raw material to the complex enzyme hydrolysis solution in step (2) is 1:2.

[0165] The enzymes in the composite enzyme hydrolyzate of step (2) are a mixture of trypsin, amylase, and cellulase, wherein the concentration of trypsin is 0.6wt%, the concentration of amylase is 0.6wt%, the concentration of cellulase is 1.2wt%, and the balance is water. The composite fermentation bacteria of step (3) are a mixture of thermophilic Streptococcus, Aspergillus niger, and Bacillus subtilis in a mass ratio of 1:1:2.

[0166] Test Example 1

[0167] Minimum inhibitory concentration (MIC) test: The bacterial enzyme peptide feed additive for preventing pathogenic stiff cattle disease and stiff sheep disease obtained in each example and comparative example and the standard example (penicillin) were prepared into aqueous solutions of different concentrations. 2 ml of the corresponding aqueous solution was taken and added to the same amount of agar medium. Salmonella was then added to the solution at a concentration of 10^6 CFU / mL. The solution was incubated at 37°C for 24 hours. The minimum inhibitory concentration (MIC) was determined by comparing the bacterial growth at different concentrations.

[0168] Table 1: Minimum inhibitory concentration test results

[0169]

[0170] Test Example 2

[0171] Stability test: The minimum inhibitory concentration (MIC) test was performed on the bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease obtained in Example 1, Example 8, Example 9, and Comparative Example 2. Then, the bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease obtained in Example 1, Example 8, Example 9, and Comparative Example 2 was placed at 25°C and left open for 7 days under the same indoor conditions. The minimum inhibitory concentration (MIC) test was performed again, and the difference between the minimum inhibitory concentration and the freshly prepared bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease was compared to reflect its stability.

[0172] Minimum inhibitory concentration (MIC) test: The bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease obtained in Example 1, Example 8, Example 9, and Comparative Example 2 was prepared into aqueous solutions of different concentrations. 2 ml of the corresponding aqueous solution was taken and added to the same amount of agar medium. Then, Salmonella was added and the concentration was adjusted to 10^6 CFU / mL. The solution was cultured at a constant temperature of 37°C for 24 hours. The minimum inhibitory concentration (MIC) was determined by comparing the bacterial growth at different concentrations.

[0173] The bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease obtained in Example 1, Example 8, Example 9, and Comparative Example 2 after being placed for 7 days was prepared into aqueous solutions of different concentrations. 2 ml of the corresponding aqueous solution was taken and added to the same amount of agar medium. Then, Salmonella was added and the concentration was adjusted to 10^6 CFU / mL. The mixture was cultured at a constant temperature of 37°C for 24 hours. The minimum inhibitory concentration (MIC) was determined by comparing the bacterial growth at different concentrations.

[0174] Table 2: Stability test results

[0175]

[0176] As can be seen from Table 1, Example 1 has the lowest minimum inhibitory concentration, and is close to the minimum inhibitory concentration of the antibiotic, which indicates that the bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease prepared in the embodiment has good inhibitory performance against Salmonella and can effectively prevent the occurrence of stiff cow disease. Example 1 of the present invention uses crushing, acid, and alkali to pretreat the raw materials to obtain a pre-enzymatic raw material; the obtained pre-enzymatic raw material and coral mucus are mixed and enzymatically hydrolyzed with a composite enzyme to obtain an enzymatic solution; then the composite fermentation bacteria are inoculated into the enzymatic solution for fermentation to obtain a fermentation liquid; finally, the fermentation liquid is spray-dried to obtain the bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease.

[0177] A comparison of Example 1 and Comparative Example 1 shows that acid-base cooking treatment can effectively improve the inhibitory performance of the prepared bacterial enzyme peptide feed additive for preventing pathogenic scurvy and scurvy against Salmonella. A comparison of Example 1 and Examples 2-4 shows that, depending on the target raw materials, selecting appropriate enzyme components and optimizing the enzymatic hydrolysis formula can improve fermentation efficiency while further enhancing the specificity of the fermentation product against scurvy.

[0178] A comparison of Example 1 and Examples 5-7 demonstrates that the combined use of Streptococcus thermophilus, Aspergillus niger, and Bacillus achieves complementary effects, leveraging the beneficial substances produced by each (such as lactic acid, antimicrobial peptides, and ethanol) to synergistically inhibit Salmonella. This synergistic effect during the fermentation process produces a wider variety of active substances, enhancing the functional diversity of the bacterial enzyme peptides.

[0179] A comparison of Examples 1, 8, and 9 demonstrates that coral mucus, as a natural source of polysaccharides and proteins, may enhance the stability of zymopeptides, particularly by protecting the membrane during the spray-drying process and preventing loss of enzyme and peptide activity. Furthermore, coral mucus may also improve the yield and quality of zymopeptides by providing nutrients and promoting microbial growth during enzymatic hydrolysis and fermentation.

[0180] As can be seen from Table 2, adding coral mucus during the enzymatic hydrolysis process has the highest stability.

[0181] Of course, the above description is only a specific embodiment of the present invention and is not intended to limit the scope of implementation of the present invention. Any equivalent changes or modifications made based on the structure, features and principles described in the scope of the patent application of the present invention should be included in the scope of the patent application of the present invention.

Claims

1. A method for preparing a bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease, characterized in that: The specific steps are as follows: (1) Pretreatment: The raw material is crushed into particles with a particle size of 1-2 mm, and the crushed raw material is added to a 0.5-1 wt % hydrochloric acid aqueous solution and acid-treated at a temperature of 90-100° C. for 1-2 hours; the acid-treated raw material is then washed with water until neutral, and a 1-2 wt % sodium hydroxide aqueous solution is added, and treated at 75-85° C. for 1-2 hours, washed with water until neutral, and dried to obtain a pre-enzymatic hydrolysis raw material; (2) adding the pre-enzymatic hydrolysis raw material and coral mucus obtained in step (1) to the compound enzyme hydrolysis solution, performing enzymatic hydrolysis at 50-60° C. for 4-6 hours, filtering to remove solid residue, and obtaining an enzymatic hydrolysis solution; (3) inoculating the composite fermentation bacteria into the enzymatic hydrolysate obtained in step (2) at an inoculation amount of 5-10 wt %, and fermenting at 30-37° C. and a pH of 4.5-5.5 for 24-48 hours. During the fermentation process, stirring was performed at a speed of 800-1000 r / min, and oxygen was supplied to maintain the dissolved oxygen content at 60-80%. After the fermentation was completed, centrifugation was performed at a speed of 800-1200 r / min for 4-6 minutes to obtain a fermentation liquid; (4) spray drying the fermentation liquid obtained in step (3) at a temperature of 60-80° C. to obtain the bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease; The raw materials in step (1) are prepared by mixing soybean meal, corn straw, wheat bran and rapeseed meal in a mass ratio of (1-2): (3-4): (2-3): (1-2); The mass ratio of the pre-enzymatic hydrolysis raw material, coral mucus, and compound enzyme hydrolyzate in step (2) is 1: (0.1-0.4): (1-2); The enzyme in the complex enzyme hydrolysis solution in step (2) is at least one of trypsin, amylase, and cellulase; The composite fermentation bacteria in step (3) is at least one of thermophilic Streptococcus, Aspergillus niger, and Bacillus subtilis.

2. The method for preparing the bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease as claimed in claim 1, characterized in that: The concentration of trypsin is 0.5-1wt%, the concentration of amylase is 0.5-1wt%, the concentration of cellulase is 1-2wt%, and the balance is water.

3. A bacterial enzyme peptide feed additive for preventing pathogenic stiff cattle disease and stiff sheep disease, characterized in that: The invention is prepared by the preparation method of the bacterial enzyme peptide feed additive for preventing pathogenic stiff cow disease and stiff sheep disease according to any one of claims 1-2.

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