Green antibiotic-free quail feed capable of promoting growth and preparation method of green antibiotic-free quail feed

Abstract

The invention discloses a green antibiotic-free quail feed capable of promoting growth and a preparation method of the green antibiotic-free quail feed, and belongs to the technical field of feed preparation. The preparation method comprises the following steps: mixing traditional Chinese medicine residues and water according to a weight part ratio of 1: (0.8-1.4), and sterilizing at 121 DEG C for 30 minutes to obtain a fermentation substrate; and mixing the fermentation substrate, the mixed strain and the mixed enzyme according to a weight part ratio of 1: (0.06-0.08): (0.008-0.009), and carrying out ventilation fermentation treatment at 30-35 DEG C for 5-7 days to obtain the green antibiotic-free quail feed. Traditional Chinese medicine residues and water are mixed to serve as a fermentation substrate, then mixed strains and mixed enzymes are inoculated into the fermentation substrate for fermentation treatment, and the green antibiotic-free quail feed with the promoting effect on quail feeding is obtained.

Description

Technical Field

[0001] This invention belongs to the field of feed preparation technology, specifically relating to a growth-promoting green antibiotic-free quail feed and its preparation method. Background Technology

[0002] Quail eggs and meat have a unique flavor and rich nutritional value, making them a popular product in the market. Safe quail farming is crucial for ensuring the supply of quail products. Currently, most feed used in the livestock industry contains additives such as antibiotics. Antibiotics are widely used in feed as growth promoters and antibacterial agents, effectively reducing livestock and poultry morbidity and improving feed conversion rates. Feed safety is the foundation of livestock and poultry product safety, directly related to human health, the ecological environment, and the sustainable development of the industry. In traditional feed production, the misuse and overuse of antibiotics is the core issue causing safety hazards. While adding antibiotics to feed can improve farming efficiency in the short term, long-term use leads to multiple safety risks. First, there is the risk of antibiotic residues. After livestock and poultry ingest feed containing antibiotics, some antibiotics cannot be completely metabolized, leaving residues in meat, eggs, and milk products. Long-term human consumption of such products may cause intestinal flora imbalance, decreased immunity, and even affect children's growth and development. Second, there is the issue of bacterial resistance. Long-term exposure to low doses of antibiotics can induce antibiotic resistance in bacteria in livestock and poultry. Once these resistant strains are transmitted to humans through the food chain, they may render clinical antibiotic treatment ineffective, posing a serious threat to public health and safety.

[0003] Antibiotic-free feed is a feed product that does not contain added antibiotics. Its core characteristic is achieving functional replacement of antibiotics through green alternatives, rather than simply removing them. Currently available antibiotic alternatives in feed include probiotics and enzyme preparations. Probiotics, also known as live microbial preparations or microecological preparations, are live microbial preparations composed of many beneficial microorganisms and their metabolites that can be directly fed to animals. They can inhibit and eliminate harmful microorganisms in the animal's gastrointestinal tract, metabolize to produce large amounts of organic acids, and lower the pH value in the gastrointestinal tract, thereby ensuring the normal flora structure of the gastrointestinal tract. When animals continuously obtain probiotics from their feed, they can maintain the absolute dominance of beneficial microorganisms in the digestive tract, maintain the stability of the animal's intestinal environment, and ensure healthy growth. Enzymes are active substances produced by organisms and act as catalysts for various biochemical reactions in the body. The digestion, absorption, and utilization of various nutrients all depend on the action of enzymes. Enzymes have characteristics such as specificity, high efficiency, and specificity. Enzyme preparations used as feed additives mainly include digestive enzymes and phytases. In the feed and animal husbandry industry, exogenous digestive enzymes can directly decompose nutrients, eliminate anti-nutritional factors, activate the secretion of endogenous enzymes, and improve the utilization rate of phytate phosphorus, thereby improving the digestibility and absorption rate of livestock and poultry feed, thus increasing feed utilization and promoting livestock and poultry growth. Microbial strains and enzymes, as important additives in the field of antibiotic-free feed, function from the perspective of feed additives. However, developing new microbial strains and enzymes is quite difficult because the structure and function of strains and enzymes are unpredictable. Therefore, it not only requires complex technical work such as sequencing and structural characterization studies, but also incurs high costs and may even lead to development failure.

[0004] Traditional Chinese medicine residue is the solid waste remaining after the extraction of effective components from traditional Chinese medicine. It mainly contains crude fiber, lignin, starch, crude protein, and residual bioactive components such as polysaccharides, alkaloids, flavonoids, saponins, and volatile oils. These advantages are further amplified through microbial fermentation, realizing multiple values ​​from waste to resources. However, directly inoculating bacterial strains into the residue for fermentation results in poor colonization due to the residue's compact structure, the presence of antibacterial components, and the presence of other bacteria, thus affecting fermentation efficiency. Therefore, improving the microbial fermentation of traditional Chinese medicine residue to prepare antibiotic-free feed that promotes quail growth is of great significance. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention uses a mixture of traditional Chinese medicine residue and water as a fermentation substrate. A mixed bacterial strain and a mixed enzyme are then inoculated into the fermentation substrate for fermentation, resulting in a green, antibiotic-free quail feed that promotes quail husbandry. Specifically, the technical solution of this invention includes the following: One objective of this invention is to provide a method for preparing a growth-promoting, green, antibiotic-free quail feed, the method comprising the following steps: The fermentation substrate is obtained by mixing Chinese herbal medicine residue and water at a weight ratio of 1:0.8~1.4 and then sterilizing at 121℃ for 30 minutes. Fermentation substrate, mixed bacterial strains and mixed enzymes were mixed in a weight ratio of 1:0.06~0.08:0.008~0.009 and then fermented at 30℃~35℃ for 5~7 days to obtain the green antibiotic-free quail feed. The moisture content of the fermentation substrate needs to be strictly controlled in this system. If the moisture content is too low, the enzymes and the fermentation substrate cannot come into full contact, and the activity of microorganisms and enzymes may be inhibited. If the moisture content is too high, the permeability of the fermentation substrate will decrease, which may easily lead to the formation of a hypoxic or even anaerobic microenvironment, resulting in an imbalance in the microbial community structure, which is not conducive to fermentation.

[0006] Furthermore, the medicinal residue is obtained by mixing and pulverizing ginseng residue, licorice residue, scutellaria baicalensis residue, poria cocos residue, isatis root residue, cassia seed residue, chuanxiong rhizome residue, and hemp seed residue in a weight ratio of 1:1:1:1:1:1 and passing them through a 50-mesh sieve. If the medicinal residue is fermented directly without sieving, it may contain large lumps, coarse fiber bundles, and localized hard lumps. In this system, this may prevent the formation of a uniform fermentation substrate system after mixing with water, which is not conducive to enzyme contact and bacterial colonization, thus affecting fermentation. On the other hand, if the medicinal residue obtained by sieving is too fine, although it is beneficial to increase the contact area, the fine residue may easily form a slurry-like mixture after mixing with water, which will damage the aeration of the fermentation system, making it easy for heat to be unable to dissipate in time, causing the bacterial enzymes to be easily inactivated, and the fine residue is not conducive to consumption.

[0007] Furthermore, the mixed strain is composed of OD 600 The Lactobacillus acidophilus bacterial solution had an OD of 0.7-0.8. 600 The concentrations of domesticated Bacillus subtilis bacterial suspension and OD were 0.6–0.7. 600 The green wood enzyme solution, with a concentration of 0.6-0.7, is composed of a weight ratio of 1:0.5-0.6:0.3-0.5. Lactobacillus acidophilus is the core strain for fermentation and acid production. It can inhibit the growth and reproduction of harmful bacteria by creating an acidic environment. If the dosage is too low, the acidity of the fermented feed will be low and the quality will be poor.

[0008] Furthermore, the OD 600 The strain of Lactobacillus acidophilus used for the 0.7~0.8% Lactobacillus acidophilus culture is CICC 6074. There are no special culture requirements; it can be cultured using conventional MRS liquid medium.

[0009] Furthermore, the method for preparing the domesticated Bacillus subtilis bacterial solution includes the following steps: The first acclimatized bacterial solution was obtained by culturing Bacillus subtilis microcapsules in a nutrient broth liquid medium with a pH of 5.0 at 28℃~30℃ for 28h~30h. The first acclimatized bacterial solution was cultured in a nutrient broth liquid medium with a pH of 4.0 at 33℃~35℃ for 24h~26h to obtain the second acclimatized bacterial solution; The second acclimatization culture was placed in a synthetic liquid medium at pH 6.0 and cultured at 35°C until OD500 reached. 600 The culture solution of domesticated Bacillus subtilis was obtained with a concentration of 0.6~0.7.

[0010] Furthermore, the preparation method of the Bacillus subtilis microcapsules includes the following steps: OD 600 A suspension dispersion was obtained by mixing Bacillus subtilis bacterial suspension (0.7-0.8), oxidized oligosaccharide aqueous dispersion, and amino polysaccharide aqueous dispersion in a weight ratio of 1:18-20:4-5 and reacting at 25°C for 12-18 hours. The suspension and sodium borohydride were mixed at a weight ratio of 1:0.01~0.02 and reacted at 20℃ for 50min~60min. After filtration and washing, Bacillus subtilis microcapsules were obtained.

[0011] Furthermore, the OD 600 The Bacillus subtilis strain used for the 0.7~0.8 Bacillus subtilis bacterial suspension is CICC 24713. There are no special culture requirements; it can be cultured in a conventional nutrient broth liquid medium.

[0012] Furthermore, the preparation method of the oxidized oligosaccharide aqueous dispersion includes the following steps: Oligosaccharides, sodium periodate, and deionized water were mixed in a weight ratio of 1:0.1~0.2:10~15 and reacted in the dark at 25°C for 5~7 hours. The mixture was then quenched, dialyzed, and freeze-dried to obtain oxidized oligosaccharides. The oxidized oligosaccharide and deionized water were mixed and dispersed at a weight ratio of 0.1:1~2 to obtain the oxidized oligosaccharide aqueous dispersion. The oligosaccharide oxidation should not be excessive, otherwise it may excessively destroy the oligosaccharide structure, resulting in too many aldehyde groups and excessive cross-linking, which is not conducive to the contact between the strain and nutrients and affects subsequent fermentation. In addition, the concentration of the oxidized oligosaccharide aqueous dispersion should not be too high, otherwise it is also easy to cause excessive cross-linking in this system, resulting in an excessively dense encapsulation structure, making it difficult for the strain to contact nutrients and exert its function.

[0013] Furthermore, the oligosaccharides include fructooligosaccharides.

[0014] Furthermore, the solvent used for quenching is ethylene glycol, the molecular weight cutoff of the dialysis bag used for dialysis is 500 Da, and the freeze-drying conditions include a temperature of -50°C, a vacuum of 10 Pa, and a drying time of 20 h.

[0015] Furthermore, the aminopolysaccharide aqueous dispersion includes a chitosan aqueous dispersion with a mass percentage concentration of 1% to 1.6%.

[0016] Furthermore, the nutrient broth liquid culture medium is obtained by adding 10g peptone, 3g beef extract powder, and 5g sodium chloride to water to a final volume of 1L, followed by sterilization at 121℃ for 20 minutes.

[0017] Furthermore, the synthetic liquid culture medium is prepared by mixing 8g-10g glucose, 5g-8g sucrose, 10g-12g peptone, 3g-5g yeast extract, 0.5g-1g L-proline, 0.2g magnesium sulfate heptahydrate, and 0.05g manganese sulfate monohydrate, bringing the volume to 1L, and then sterilizing at 121°C for 20 minutes. During the acid acclimatization process of this invention, the strain may accumulate proline intracellularly to resist acid stress. Exogenous addition of L-proline can reduce the strain's own synthetic energy consumption, accelerate activity recovery, and solidify acid tolerance. Moreover, the acid tolerance acclimatization of this invention may lead to a decrease in the activity of Bacillus subtilis' stress-resistant enzymes and other active enzymes. Supplementing with Mg... 2+ and Mn 2+ Activating enzyme activity enhances bacterial metabolism. However, further increases in proline, magnesium, and manganese ions may disrupt the osmotic fluid and ion balance, potentially leading to bacterial inactivation.

[0018] Furthermore, the OD 600 The strain of green wood enzyme used for the green wood enzyme culture with a concentration of 0.6~0.7 is CICC 13046. There are no special culture requirements, and it can be cultured in conventional PDA liquid medium.

[0019] Furthermore, the mixed enzyme is composed of cellulase, xylanase and pectinase in a weight ratio of 1:0.2~0.3:0.6~0.8.

[0020] Furthermore, the ventilation rate for the ventilated fermentation is 0.3L / min to 0.4L / min.

[0021] The second objective of this invention is to provide a green, antibiotic-free quail feed that promotes growth.

[0022] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention uses traditional Chinese medicine residue as a raw material for antibiotic-free fermented feed for quail. Lactic acid bacteria, Bacillus subtilis, and viridinium chlorophyll are used as mixed bacterial strains for fermenting the residue. Lactic acid bacteria not only lower the pH of the fermentation system, inhibiting the growth of harmful bacteria, but also impart a mild sour aroma to the feed, masking the bitterness and off-flavor of the residue and increasing animal appetite. Viridinium chlorophyll degrades a large amount of crude fiber, facilitating chewing and digestion. Bacillus subtilis secretes antibacterial substances such as lipopeptides and bacitracin, further enhancing the inhibition of harmful bacteria. Cellulase, xylanase, and pectinase are used as mixed enzymes for synergistic fermentation. These enzymes promote the degradation of cell walls in the residue, releasing more nutrients and increasing the nutrient content of the fermentation substrate, thus indirectly promoting the fermentation process. Through the synergistic effect of bacteria and enzymes in fermenting the residue, the resulting feed has a positive effect on quail farming. However, in actual breeding, it was found that the egg production rate and weight gain of quails were poor. This may be because the acidity in the system during feed fermentation was too strong, which inhibited the metabolic activity of Bacillus subtilis to a certain extent, thus affecting the fermentation process. Therefore, acid tolerance acclimation was carried out on Bacillus subtilis to increase its adaptability to acidic environments. However, it was found that the acclimated Bacillus subtilis cultured directly and then used for feeding with fermented Chinese medicine residues performed poorly. This may be because the strain was directly placed in an acidic culture environment for acclimation, and the early acid stress may have damaged the homeostasis of the strain's cells, making the strain unable to adapt to the excessively acidic system and greatly reducing its survival rate. Therefore, oligosaccharides, which can provide nutrients for the strain and thus promote its proliferation, were used as one of the encapsulation carriers. Oxidation was performed to break the ortho-dihydroxy groups in the structure of the oligosaccharide, producing aldehyde groups. Subsequently, with the help of... Amino polysaccharides can cross-link with amino groups, thereby encapsulating and protecting the strain. This encapsulation helps reduce the impact of the strain losing its activity too quickly due to direct contact with the acidic culture system. However, since the imine bond formed by the cross-linking of amino and aldehyde groups has an acid hydrolysis effect, the acidic acclimatization environment will cause the carrier to decompose and lose its protective effect. Therefore, by reducing the imine bond to form a stable secondary amine structure that is resistant to acid hydrolysis, the protective stability of the carrier is improved, allowing it to be steadily acclimatized in an acidic environment. This improves the activity of the acclimatized Bacillus subtilis cultured after acclimatization, which can improve the feeding effect of the feed when it is subsequently used for fermentation to prepare feed. Detailed Implementation

[0023] The technical solution of the present invention will be clearly and completely described below through embodiments. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of them. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0024] Unless otherwise stated, all raw materials and reagents used in this invention are commercially available or can be prepared by known methods.

[0025] Preparation Example 1 The preparation process of Chinese herbal medicine residue is as follows: Weigh out equal parts of ginseng residue, licorice residue, scutellaria residue, poria residue, isatis root residue, cassia seed residue, chuanxiong root residue, and hemp seed residue, mix them together, and place them in a 55℃ oven to dry and remove moisture for 24 hours. After drying, remove them and pulverize them with a pulverizer until they can pass through a 50-mesh sieve to obtain the Chinese herbal medicine residue.

[0026] Preparation Example 2 The preparation process of Chinese herbal medicine residue is as follows: Weigh out equal parts of ginseng residue, licorice residue, scutellaria residue, poria residue, isatis root residue, cassia seed residue, chuanxiong residue, and hemp seed residue, mix them together, and place them in a 55℃ oven to dry and remove moisture for 24 hours before using them directly as Chinese medicine residue.

[0027] Preparation Example 3 The preparation process of Chinese herbal medicine residue is as follows: Weigh out equal parts of ginseng residue, licorice residue, scutellaria residue, poria residue, isatis root residue, cassia seed residue, chuanxiong root residue, and hemp seed residue, mix them together, and place them in a 55℃ oven to dry and remove moisture for 24 hours. After drying, remove them and pulverize them with a pulverizer until they can pass through a 200-mesh sieve to obtain the Chinese herbal medicine residue.

[0028] Preparation Example 4 The preparation process of the oxidized oligosaccharide aqueous dispersion is as follows: Weigh 1 part by weight of fructooligosaccharide and 10 parts by weight of deionized water, heat and stir until dissolved and homogeneous. Then, maintain a rotation speed of 200 r / min and cool to 25°C. Add 0.1 part by weight of sodium periodate, mix immediately, and protect from light. Stir and react for 5 hours. After the reaction is complete, add 1 part by weight of ethylene glycol and continue stirring for half an hour to quench the sodium periodate. Pour the resulting mixture into a dialysis bag with a molecular weight cutoff of 500 Da and dialyze with distilled water for 2 days. After dialysis, transfer the contents of the dialysis bag to a freeze dryer, first evacuate to 10 Pa, then cool to -50°C and dry for 20 hours to obtain oxidized oligosaccharide. Weigh 0.1 part by weight of oxidized oligosaccharide and 1 part by weight of deionized water, mix and disperse evenly to form an oxidized oligosaccharide aqueous dispersion.

[0029] Preparation Example 5 The preparation process of the oxidized oligosaccharide aqueous dispersion is as follows: Weigh 1 part by weight of fructooligosaccharide and 15 parts by weight of deionized water, heat and stir until dissolved and homogeneous. Then, maintain a rotation speed of 200 r / min and cool to 25°C. Add 0.2 parts by weight of sodium periodate, mix immediately, and protect from light. Stir and react for 7 hours. After the reaction is complete, add 1 part by weight of ethylene glycol and continue stirring for half an hour to quench the sodium periodate. Pour the resulting mixture into a dialysis bag with a molecular weight cutoff of 500 Da and dialyze with distilled water for 2 days. After dialysis, transfer the contents of the dialysis bag to a freeze dryer, first evacuate to 10 Pa, then cool to -50°C and dry for 20 hours to obtain oxidized oligosaccharide. Weigh 0.1 parts by weight of oxidized oligosaccharide and 2 parts by weight of deionized water, mix and disperse evenly to form an oxidized oligosaccharide aqueous dispersion.

[0030] Preparation Example 6 The preparation process of the oxidized oligosaccharide aqueous dispersion is as follows: Weigh 1 part by weight of fructooligosaccharide and 15 parts by weight of deionized water, heat and stir until dissolved and homogeneous. Then, maintain a rotation speed of 200 r / min and cool to 25°C. Add 0.5 parts by weight of sodium periodate, mix immediately, and protect from light. Stir and react for 8 hours. After the reaction is complete, add 1 part by weight of ethylene glycol and continue stirring for half an hour to quench the sodium periodate. Pour the resulting mixture into a dialysis bag with a molecular weight cutoff of 500 Da and dialyze with distilled water for 2 days. After dialysis, transfer the contents of the dialysis bag to a freeze dryer, first evacuate to 10 Pa, then cool to -50°C and dry for 20 hours to obtain oxidized oligosaccharide. Weigh 0.1 parts by weight of oxidized oligosaccharide and 2 parts by weight of deionized water, mix and disperse evenly to form an oxidized oligosaccharide aqueous dispersion.

[0031] Preparation Example 7 The preparation process of the oxidized oligosaccharide aqueous dispersion is as follows: Weigh 1 part by weight of the oxidized oligosaccharide obtained in Preparation Example 5 and 2 parts by weight of deionized water, mix and disperse evenly to form an oxidized oligosaccharide aqueous dispersion.

[0032] Preparation Example 8 The preparation process of Bacillus subtilis microcapsules is as follows: Take 1 part by weight of OD obtained from Bacillus subtilis strain number CICC 24713. 600A suspension was prepared by mixing 0.7 parts by weight of Bacillus subtilis bacterial culture, 18 parts by weight of the oxidized oligosaccharide aqueous dispersion obtained in Preparation Example 4, and 4 parts by weight of the chitosan aqueous dispersion with a mass percentage concentration of 1%. The mixture was stirred and placed at 25°C for 12 hours to allow cross-linking reaction, resulting in a suspension. One part by weight of the suspension was weighed and cooled to 20°C. Then, 0.01 parts by weight of sodium borohydride was added and the mixture was stirred for 50 minutes. After the reaction, the pH of the mixture was adjusted to neutral. The solid particles were collected by vacuum filtration, washed with phosphate buffer solution at pH 6.8, washed with deionized water, and finally filtered again to obtain Bacillus subtilis microcapsules.

[0033] Preparation Example 9 The preparation process of Bacillus subtilis microcapsules is as follows: Take 1 part by weight of OD obtained from Bacillus subtilis strain number CICC 24713. 600 A suspension was prepared by mixing 0.7 parts by weight of Bacillus subtilis bacterial culture, 19 parts by weight of the oxidized oligosaccharide aqueous dispersion obtained in Preparation Example 4, and 4 parts by weight of the chitosan aqueous dispersion with a mass percentage concentration of 1.2%. The mixture was stirred and placed at 25°C for 14 hours to allow cross-linking reaction, resulting in a suspension. One part by weight of the suspension was weighed and cooled to 20°C. Then, 0.01 parts by weight of sodium borohydride was added and the mixture was stirred for 50 minutes. After the reaction, the pH of the mixture was adjusted to neutral. The solid particles were collected by vacuum filtration, washed with phosphate buffer solution at pH 6.8, washed with deionized water, and finally filtered again to obtain Bacillus subtilis microcapsules.

[0034] Preparation Example 10 The preparation process of Bacillus subtilis microcapsules is as follows: Take 1 part by weight of OD obtained from Bacillus subtilis strain number CICC 24713. 600 A suspension was prepared by mixing 0.8 parts by weight of Bacillus subtilis bacterial culture, 19 parts by weight of the oxidized oligosaccharide aqueous dispersion obtained in Preparation Example 5, and 5 parts by weight of the chitosan aqueous dispersion with a mass percentage concentration of 1.4%. The mixture was stirred and placed at 25°C for 16 hours to allow cross-linking reaction, resulting in a suspension. One part by weight of the suspension was weighed and cooled to 20°C. Then, 0.02 parts by weight of sodium borohydride was added and the mixture was stirred for 55 minutes. After the reaction, the pH of the mixture was adjusted to neutral. Solid particles were collected by vacuum filtration, followed by washing with phosphate buffer solution at pH 6.8, then washing with deionized water, and finally filtration to obtain Bacillus subtilis microcapsules.

[0035] Preparation Example 11 The preparation process of Bacillus subtilis microcapsules is as follows: Take 1 part by weight of OD obtained from Bacillus subtilis strain number CICC 24713.600 A suspension was prepared by mixing 0.8 parts by weight of Bacillus subtilis bacterial culture, 20 parts by weight of the oxidized oligosaccharide aqueous dispersion obtained in Preparation Example 5, and 5 parts by weight of the chitosan aqueous dispersion with a mass percentage concentration of 1.6%. The mixture was stirred and placed at 25°C for 18 hours to allow cross-linking reaction, resulting in a suspension. One part by weight of the suspension was weighed and cooled to 20°C. Then, 0.02 parts by weight of sodium borohydride was added and the mixture was stirred for 60 minutes. After the reaction, the pH of the mixture was adjusted to neutral. Solid particles were collected by vacuum filtration, followed by washing with phosphate buffer solution at pH 6.8, then washing with deionized water, and finally filtration to obtain Bacillus subtilis microcapsules.

[0036] Preparation Example 12 The preparation process of Bacillus subtilis microcapsules is as follows: The oxidized oligosaccharide aqueous dispersion in Preparation Example 11 was replaced with the oxidized oligosaccharide aqueous dispersion obtained in Preparation Example 6, and the rest of the preparation process was the same as in Preparation Example 11.

[0037] Preparation Example 13 The preparation process of Bacillus subtilis microcapsules is as follows: The oxidized oligosaccharide aqueous dispersion in Preparation Example 11 was replaced with the oxidized oligosaccharide aqueous dispersion obtained in Preparation Example 7, and the rest of the preparation process was the same as in Preparation Example 11.

[0038] Preparation Example 14 The preparation process of Bacillus subtilis microcapsules is as follows: Take 1 part by weight of OD obtained from Bacillus subtilis strain number CICC 24713. 600 0.8% Bacillus subtilis bacterial culture, 20 parts by weight of the oxidized oligosaccharide aqueous dispersion obtained in Preparation Example 5, and 5 parts by weight of the chitosan aqueous dispersion with a mass percentage concentration of 1.6% were mixed and stirred and placed in 25°C for 18 hours to undergo cross-linking reaction to obtain a suspension dispersion; the solid particles were collected by vacuum filtration, then washed with phosphate buffer at pH 6.8, then washed with deionized water, and finally filtered again to obtain Bacillus subtilis microcapsules.

[0039] Preparation Example 15 The preparation process of the nutrient broth liquid culture medium is as follows: Weigh 10g of peptone, 3g of beef extract powder and 5g of sodium chloride, add water to make up to 1L, sterilize in a sterilizer at 121℃ for 20min and then cool to room temperature.

[0040] Preparation Example 16 The preparation process of synthetic liquid culture medium is as follows: Weigh out 8g glucose, 5g sucrose, 10g peptone, 3g yeast extract, 0.5g L-proline, 0.2g magnesium sulfate heptahydrate and 0.05g manganese sulfate monohydrate, mix and bring the volume to 1L, then sterilize in a sterilizer at 121℃ for 20 minutes and cool to room temperature.

[0041] Preparation Example 17 The preparation process of synthetic liquid culture medium is as follows: Weigh out 10g glucose, 8g sucrose, 12g peptone, 5g yeast extract, 1g L-proline, 0.2g magnesium sulfate heptahydrate and 0.05g manganese sulfate monohydrate, mix and bring the volume to 1L, then sterilize in a sterilizer at 121℃ for 20 minutes and cool to room temperature.

[0042] Preparation Example 18 The preparation process of synthetic liquid culture medium is as follows: Weigh out 10g glucose, 8g sucrose, 12g peptone, 5g yeast extract, 2g L-proline, 0.5g magnesium sulfate heptahydrate and 0.1g manganese sulfate monohydrate, mix and bring the volume to 1L, then sterilize in a sterilizer at 121℃ for 20min and cool to room temperature.

[0043] Preparation Example 19 The preparation process of domesticated Bacillus subtilis bacterial culture is as follows: On a clean bench, the pH of the nutrient broth liquid culture medium obtained in Preparation Example 15 was adjusted to 5.0 with acetic acid. Then, Bacillus subtilis microcapsules obtained in Preparation Example 8 were added to the nutrient broth liquid culture medium at pH 5.0 at 10% of the total culture medium volume and mixed. The mixture was then placed in a shaker at 28°C and cultured at 150 rpm for 28 hours to obtain the first acclimatized bacterial solution. Separately, the pH of the nutrient broth liquid culture medium obtained in Preparation Example 15 was adjusted to 4.0 with acetic acid. Then, Bacillus subtilis microcapsules obtained in Preparation Example 8 were added to the nutrient broth liquid culture medium at pH 5.0 at 10% of the total culture medium volume and mixed. The first acclimatized bacterial solution was added to a nutrient broth liquid medium with a pH of 4.0 and mixed. The mixture was then placed in a shaker at 33°C and cultured at 150 rpm for 24 hours to obtain the second acclimatized bacterial solution. The pH of the synthetic liquid medium obtained in Preparation Example 16 was adjusted to 6.0 with acetic acid. Then, 10% of the weight of the second acclimatized bacterial solution was added to the synthetic liquid medium at pH 6.0 and mixed. The mixture was then placed in a shaker at 35°C and cultured at 150 rpm until the OD of the bacterial solution reached a certain level. 600 The concentration was 0.6, completing the preparation of the domesticated Bacillus subtilis bacterial solution.

[0044] Preparation Example 20 The preparation process of domesticated Bacillus subtilis bacterial culture is as follows: On a clean bench, the pH of the nutrient broth liquid culture medium obtained in Preparation Example 15 was adjusted to 5.0 with acetic acid. Then, Bacillus subtilis microcapsules obtained in Preparation Example 9 were added to the nutrient broth liquid culture medium at pH 5.0 at 10% of the total culture medium volume and mixed. The mixture was then placed in a shaker at 28°C and cultured at 150 rpm for 28 hours to obtain the first acclimatized bacterial solution. Separately, the pH of the nutrient broth liquid culture medium obtained in Preparation Example 15 was adjusted to 4.0 with acetic acid. Then, Bacillus subtilis microcapsules obtained in Preparation Example 9 were added to the nutrient broth liquid culture medium at pH 5.0 at 10% of the total culture medium volume and mixed. The first acclimatized bacterial solution was added to a nutrient broth liquid medium with a pH of 4.0 and mixed. The mixture was then placed in a shaker at 33°C and cultured at 150 rpm for 24 hours to obtain the second acclimatized bacterial solution. The pH of the synthetic liquid medium obtained in Preparation Example 16 was adjusted to 6.0 with acetic acid. Then, 10% of the weight of the second acclimatized bacterial solution was added to the synthetic liquid medium at pH 6.0 and mixed. The mixture was then placed in a shaker at 35°C and cultured at 150 rpm until the OD of the bacterial solution reached a certain level. 600 The concentration was 0.6, completing the preparation of the domesticated Bacillus subtilis bacterial solution.

[0045] Preparation Example 21 The preparation process of domesticated Bacillus subtilis bacterial culture is as follows: On a clean bench, the pH of the nutrient broth liquid culture medium obtained in Preparation Example 15 was adjusted to 5.0 with acetic acid. Then, Bacillus subtilis microcapsules obtained in Preparation Example 10 were added to the nutrient broth liquid culture medium at pH 5.0 at 10% of the total culture medium volume and mixed. The mixture was then placed in a shaker at 30°C and cultured at 150 rpm for 30 hours to obtain the first acclimatized bacterial solution. Separately, the pH of the nutrient broth liquid culture medium obtained in Preparation Example 15 was adjusted to 4.0 with acetic acid. Then, Bacillus subtilis microcapsules obtained in Preparation Example 10 were added to the nutrient broth liquid culture medium at pH 5.0 at 10% of the total culture medium volume. 0%, the first acclimatized bacterial solution was added to a nutrient broth liquid medium with a pH of 4.0 and mixed, then placed in a shaker at 35°C and cultured at a speed of 150 r / min for 26 h to obtain the second acclimatized bacterial solution; the pH of the synthetic liquid medium obtained in Preparation Example 17 was adjusted to 6.0 with acetic acid, and then 10% of the weight of the medium was added to the synthetic liquid medium with a pH of 6.0 and mixed, then placed in a shaker at 35°C and cultured at a speed of 150 r / min until the OD of the bacterial solution reached 0%. 600 The concentration was 0.7, completing the preparation of the domesticated Bacillus subtilis bacterial solution.

[0046] Preparation Example 22 The preparation process of domesticated Bacillus subtilis bacterial culture is as follows: On a clean bench, the pH of the nutrient broth liquid culture medium obtained in Preparation Example 15 was adjusted to 5.0 with acetic acid. Then, Bacillus subtilis microcapsules obtained in Preparation Example 11 were added to the nutrient broth liquid culture medium at pH 5.0 at 10% of the total culture medium volume and mixed. The mixture was then placed in a shaker at 30°C and cultured at 150 r / min for 30 h to obtain the first acclimatized bacterial solution. Separately, the pH of the nutrient broth liquid culture medium obtained in Preparation Example 15 was adjusted to 4.0 with acetic acid. Then, Bacillus subtilis microcapsules obtained in Preparation Example 11 were added to the nutrient broth liquid culture medium at pH 5.0 at 10% of the total culture medium volume. 0%, the first acclimatized bacterial solution was added to a nutrient broth liquid medium with a pH of 4.0 and mixed, then placed in a shaker at 35°C and cultured at a speed of 150 r / min for 26 h to obtain the second acclimatized bacterial solution; the pH of the synthetic liquid medium obtained in Preparation Example 17 was adjusted to 6.0 with acetic acid, and then 10% of the weight of the medium was added to the synthetic liquid medium with a pH of 6.0 and mixed, then placed in a shaker at 35°C and cultured at a speed of 150 r / min until the OD of the bacterial solution reached 0%. 600 The concentration was 0.7, completing the preparation of the domesticated Bacillus subtilis bacterial solution.

[0047] Preparation Example 23 The preparation process of domesticated Bacillus subtilis bacterial culture is as follows: The Bacillus subtilis microcapsules in Preparation Example 22 were replaced with the Bacillus subtilis microcapsules obtained in Preparation Example 12, and the rest of the preparation process was the same as in Preparation Example 22.

[0048] Preparation Example 24 The preparation process of domesticated Bacillus subtilis bacterial culture is as follows: The Bacillus subtilis microcapsules in Preparation Example 22 were replaced with the Bacillus subtilis microcapsules obtained in Preparation Example 13, and the rest of the preparation process was the same as in Preparation Example 22.

[0049] Preparation Example 25 The preparation process of domesticated Bacillus subtilis bacterial culture is as follows: The Bacillus subtilis microcapsules in Preparation Example 22 were replaced with the Bacillus subtilis microcapsules obtained in Preparation Example 14, and the rest of the preparation process was the same as in Preparation Example 22.

[0050] Preparation Example 26 The preparation process of domesticated Bacillus subtilis bacterial culture is as follows: The synthetic liquid culture medium in Preparation Example 22 was replaced with the synthetic liquid culture medium obtained in Preparation Example 18, and the rest of the preparation process was the same as in Preparation Example 22.

[0051] Preparation Example 27 The preparation process of domesticated Bacillus subtilis bacterial culture is as follows: The Bacillus subtilis microcapsules in Preparation Example 22 were replaced with OD 600 The concentration of Bacillus subtilis bacterial culture was 0.8, and the rest of the preparation process was the same as in Preparation Example 22.

[0052] Preparation Example 28 The preparation process of domesticated Bacillus subtilis bacterial culture is as follows: On a clean bench, the pH of the nutrient broth liquid culture medium obtained in Preparation Example 15 was adjusted to 5.0 with acetic acid. Then, Bacillus subtilis microcapsules obtained in Preparation Example 11 were added to the nutrient broth liquid culture medium at pH 4.0, accounting for 10% of the total culture medium weight. The mixture was then placed in a shaker at 30°C and cultured at 150 rpm for 30 h to obtain the acclimatized bacterial solution. The pH of the synthetic liquid culture medium obtained in Preparation Example 17 was adjusted to 6.0 with acetic acid. Then, the acclimatized bacterial solution was added to the synthetic liquid culture medium at pH 6.0, accounting for 10% of the culture medium weight. The mixture was then placed in a shaker at 35°C and cultured at 150 rpm until the OD of the bacterial solution reached a certain level. 600 The concentration was 0.7, completing the preparation of the domesticated Bacillus subtilis bacterial solution.

[0053] Preparation Example 29 The preparation process of the mixed strain is as follows: Weigh 1 part by weight of OD obtained from the culture of Lactobacillus acidophilus strain number CICC 6074. 600 The OD obtained in Example 19 was prepared by using 0.7 parts by weight of Lactobacillus acidophilus bacterial culture and 0.5 parts by weight. 600 The OD was obtained by culturing 0.6 parts of domesticated Bacillus subtilis culture and 0.3 parts by weight of green xylem enzyme cultured with strain number CICC 13046. 600 The green wood enzyme culture solution at 0.6 is mixed and stored for later use.

[0054] Preparation Example 30 The preparation process of the mixed strain is as follows: Weigh 1 part by weight of OD obtained from the culture of Lactobacillus acidophilus strain number CICC 6074. 600 The OD obtained in Example 20 was prepared by using 0.7 parts by weight of Lactobacillus acidophilus bacterial culture and 0.5 parts by weight. 600 The OD was obtained by culturing 0.6 parts of domesticated Bacillus subtilis culture and 0.3 parts by weight of green xylem enzyme cultured with strain number CICC 13046. 600 The green wood enzyme culture solution at 0.6 is mixed and stored for later use.

[0055] Preparation Example 31 The preparation process of the mixed strain is as follows: Weigh 1 part by weight of OD obtained from the culture of Lactobacillus acidophilus strain number CICC 6074.600 The OD obtained in Example 21 was prepared by using 0.8 parts by weight of Lactobacillus acidophilus bacterial culture and 0.6 parts by weight. 600 The OD was obtained by culturing 0.7 parts of domesticated Bacillus subtilis culture and 0.4 parts by weight of green xylem enzyme culture with strain number CICC 13046. 600 The green wood enzyme culture solution at 0.7 is mixed and stored for later use.

[0056] Preparation Example 32 The preparation process of the mixed strain is as follows: Weigh 1 part by weight of OD obtained from the culture of Lactobacillus acidophilus strain number CICC 6074. 600 The OD obtained in Example 22 was prepared by using 0.8 parts by weight of Lactobacillus acidophilus bacterial culture and 0.6 parts by weight. 600 The OD was obtained by culturing 0.7 parts of domesticated Bacillus subtilis culture and 0.5 parts by weight of green xylem enzyme cultured with strain number CICC 13046. 600 The green wood enzyme culture solution at 0.7 is mixed and stored for later use.

[0057] Preparation Example 33 The preparation process of the mixed strain is as follows: The domesticated Bacillus subtilis culture in Preparation Example 32 was replaced with the domesticated Bacillus subtilis culture obtained in Preparation Example 23, and the rest of the preparation process was the same as in Preparation Example 32.

[0058] Preparation Example 34 The preparation process of the mixed strain is as follows: The domesticated Bacillus subtilis culture in Preparation Example 32 was replaced with the domesticated Bacillus subtilis culture obtained in Preparation Example 24, and the rest of the preparation process was the same as in Preparation Example 32.

[0059] Preparation Example 35 The preparation process of the mixed strain is as follows: The domesticated Bacillus subtilis culture in Preparation Example 32 was replaced with the domesticated Bacillus subtilis culture obtained in Preparation Example 25, and the rest of the preparation process was the same as in Preparation Example 32.

[0060] Preparation Example 36 The preparation process of the mixed strain is as follows: The domesticated Bacillus subtilis culture in Preparation Example 32 was replaced with the domesticated Bacillus subtilis culture obtained in Preparation Example 26, and the rest of the preparation process was the same as in Preparation Example 32.

[0061] Preparation Example 37 The preparation process of the mixed strain is as follows: The domesticated Bacillus subtilis culture in Preparation Example 32 was replaced with the domesticated Bacillus subtilis culture obtained in Preparation Example 27, and the rest of the preparation process was the same as in Preparation Example 32.

[0062] Preparation Example 38 The preparation process of the mixed strain is as follows: The domesticated Bacillus subtilis culture in Preparation Example 32 was replaced with the domesticated Bacillus subtilis culture obtained in Preparation Example 28, and the rest of the preparation process was the same as in Preparation Example 32.

[0063] Preparation Example 39 The preparation process of the mixed strain is as follows: The domesticated Bacillus subtilis culture in Preparation Example 32 was replaced with OD. 600 The concentration of Bacillus subtilis bacterial culture was 0.8, and the rest of the preparation process was the same as in Preparation Example 32.

[0064] Preparation Example 40 The preparation process of the mixed strain is as follows: The amount of Lactobacillus acidophilus bacterial solution used in Preparation Example 32 was reduced to 0.5 parts by weight, while the rest of the preparation process remained the same as in Preparation Example 32.

[0065] Example 1

[0066] A method for preparing a growth-promoting, green, antibiotic-free quail feed specifically includes the following steps: Weigh out parts by weight of the Chinese herbal medicine residue obtained in Preparation Example 1 and 0.8 parts by weight of deionized water, mix and stir evenly, and then put it into a sterilization device at 121℃ for 30 minutes to obtain fermentation substrate; weigh out 1 part by weight of fermentation substrate, 0.06 parts by weight of mixed strain obtained in Preparation Example 29 and 0.008 parts by weight of mixed enzyme (composed of cellulase, xylanase and pectinase in a weight ratio of 1:0.2:0.6) and mix them in a fermenter, then control the air flow rate to 0.3L / min and ventilate fermentation at 30℃ for 5 days to obtain green antibiotic-free quail feed.

[0067] Example 2

[0068] A method for preparing a growth-promoting, green, antibiotic-free quail feed specifically includes the following steps: Weigh out parts by weight of the Chinese herbal medicine residue obtained in Preparation Example 1 and mix it with 1 part by weight of deionized water. Stir the mixture evenly and then sterilize it in a sterilizer at 121°C for 30 minutes to obtain the fermentation substrate. Weigh out 1 part by weight of the fermentation substrate, 0.06 parts by weight of the mixed strain obtained in Preparation Example 30, and 0.008 parts by weight of the mixed enzyme (composed of cellulase, xylanase, and pectinase in a weight ratio of 1:0.2:0.7) and mix them in a fermenter. Then control the air flow rate to 0.35 L / min and ventilate the fermentation at 32°C for 5 days to obtain green antibiotic-free quail feed.

[0069] Example 3

[0070] A method for preparing a growth-promoting, green, antibiotic-free quail feed specifically includes the following steps: Weigh 1.2 parts by weight of the Chinese herbal medicine residue obtained in Preparation Example 1, mix and stir evenly, and then put it into a sterilization device at 121℃ for 30 minutes to obtain fermentation substrate; weigh 1 part by weight of fermentation substrate, 0.07 parts by weight of the mixed strain obtained in Preparation Example 31 and 0.009 parts by weight of mixed enzyme (composed of cellulase, xylanase and pectinase in a weight ratio of 1:0.3:0.7) and mix them in a fermenter, then control the air flow rate to 0.35L / min and ventilate fermentation at 34℃ for 6 days to obtain green antibiotic-free quail feed.

[0071] Example 4

[0072] A method for preparing a growth-promoting, green, antibiotic-free quail feed specifically includes the following steps: Weigh out parts by weight of the Chinese herbal medicine residue obtained in Preparation Example 1 and mix it with 1.4 parts by weight of deionized water. Stir the mixture evenly and then sterilize it in a sterilizer at 121°C for 30 minutes to obtain the fermentation substrate. Weigh out 1 part by weight of the fermentation substrate, 0.08 parts by weight of the mixed strain obtained in Preparation Example 32, and 0.009 parts by weight of the mixed enzyme (composed of cellulase, xylanase, and pectinase in a weight ratio of 1:0.3:0.8) and mix them in a fermenter. Then control the air flow rate to 0.4 L / min and ventilate the fermentation at 35°C for 7 days to obtain green antibiotic-free quail feed.

[0073] Comparative Example 1 A method for preparing a growth-promoting, green, antibiotic-free quail feed specifically includes the following steps: Weigh out parts by weight of the Chinese herbal medicine residue obtained in Preparation Example 1 and 0.5 parts by weight of deionized water, mix and stir evenly, then put it into a sterilization device at 121℃ for 30 minutes to obtain fermentation substrate; weigh out 1 part by weight of fermentation substrate, 0.08 parts by weight of mixed strain obtained in Preparation Example 32 and 0.009 parts by weight of mixed enzyme (composed of cellulase, xylanase and pectinase in a weight ratio of 1:0.3:0.8) and mix them in a fermenter, then control the air flow rate to 0.4 L / min and ventilate fermentation at 35℃ for 7 days to obtain green antibiotic-free quail feed.

[0074] Comparative Example 2 A method for preparing a growth-promoting, green, antibiotic-free quail feed specifically includes the following steps: Weigh out parts by weight of the Chinese herbal medicine residue obtained in Preparation Example 1 and 2 parts by weight of deionized water, mix and stir evenly, then put it into a sterilization device at 121℃ for 30 minutes to obtain fermentation substrate; weigh out 1 part by weight of fermentation substrate, 0.08 parts by weight of mixed strain obtained in Preparation Example 32 and 0.009 parts by weight of mixed enzyme (composed of cellulase, xylanase and pectinase in a weight ratio of 1:0.3:0.8) and mix them in a fermenter, then control the air flow rate to 0.4L / min and ventilate fermentation at 35℃ for 7 days to obtain green antibiotic-free quail feed.

[0075] Comparative Example 3 A method for preparing a growth-promoting, green, antibiotic-free quail feed specifically includes the following steps: The Chinese herbal residue in Example 4 was replaced with the Chinese herbal residue obtained in Preparation Example 2, and the rest of the preparation process was the same as in Example 4.

[0076] Comparative Example 4 A method for preparing a growth-promoting, green, antibiotic-free quail feed specifically includes the following steps: The Chinese herbal residue in Example 4 was replaced with the Chinese herbal residue obtained in Preparation Example 3, and the rest of the preparation process was the same as in Example 4.

[0077] Comparative Example 5 A method for preparing a growth-promoting, green, antibiotic-free quail feed specifically includes the following steps: The mixed strain in Example 4 was replaced with the mixed strain obtained in Preparation Example 33, and the rest of the preparation process was the same as in Example 4.

[0078] Comparative Example 6 A method for preparing a growth-promoting, green, antibiotic-free quail feed specifically includes the following steps: The mixed strain in Example 4 was replaced with the mixed strain obtained in Preparation Example 34, and the rest of the preparation process was the same as in Example 4.

[0079] Comparative Example 7 A method for preparing a growth-promoting, green, antibiotic-free quail feed specifically includes the following steps: The mixed strain in Example 4 was replaced with the mixed strain obtained in Preparation Example 35, and the rest of the preparation process was the same as in Example 4.

[0080] Comparative Example 8 A method for preparing a growth-promoting, green, antibiotic-free quail feed specifically includes the following steps: The mixed strain in Example 4 was replaced with the mixed strain obtained in Preparation Example 36, and the rest of the preparation process was the same as in Example 4.

[0081] Comparative Example 9 A method for preparing a growth-promoting, green, antibiotic-free quail feed specifically includes the following steps: The mixed strain in Example 4 was replaced with the mixed strain obtained in Preparation Example 37, and the rest of the preparation process was the same as in Example 4.

[0082] Comparative Example 10 A method for preparing a growth-promoting, green, antibiotic-free quail feed specifically includes the following steps: The mixed strain in Example 4 was replaced with the mixed strain obtained in Preparation Example 38, and the rest of the preparation process was the same as in Example 4.

[0083] Comparative Example 11 A method for preparing a growth-promoting, green, antibiotic-free quail feed specifically includes the following steps: The mixed strain in Example 4 was replaced with the mixed strain obtained in Preparation Example 39, and the rest of the preparation process was the same as in Example 4.

[0084] Comparative Example 12 A method for preparing a growth-promoting, green, antibiotic-free quail feed specifically includes the following steps: The mixed enzyme in Example 4 was removed and not added, while the rest of the preparation process remained the same as in Example 4.

[0085] Comparative Example 13 A method for preparing a growth-promoting, green, antibiotic-free quail feed specifically includes the following steps: The amount of mixed enzyme in Example 4 was increased to 0.18 parts by weight, while the rest of the preparation process remained the same as in Example 4.

[0086] Comparative Example 14 A method for preparing a growth-promoting, green, antibiotic-free quail feed specifically includes the following steps: The mixed strain in Example 4 was replaced with the mixed strain obtained in Preparation Example 40, and the rest of the preparation process was the same as in Example 4.

[0087] Observe whether the feed obtained in Examples 1-4 and Comparative Examples 1-14 shows signs of mold growth and record the odor. Then weigh 20g of the feed obtained in Examples 1-4 and Comparative Examples 1-14 respectively, add 180mL of deionized water and shake thoroughly for 30min. Measure the pH with a pH meter in an environment of 25℃±2℃. The results are shown in Table 1 below.

[0088] Table 1 Feed Quality Source of materials Is it moldy? odor pH Example 1 no Light sour aroma 4.15 Example 2 no Light sour aroma 4.11 Example 3 no Strong sour aroma 4.08 Example 4 no Strong sour aroma 4.06 Comparative Example 1 no The smell is slightly pungent. 4.56 Comparative Example 2 no The smell is slightly pungent. 4.64 Comparative Example 3 no The smell is slightly pungent. 4.73 Comparative Example 4 yes The pungent odor is obvious. 5.13 Comparative Example 5 yes The pungent odor is obvious. 5.34 Comparative Example 6 yes The pungent odor is obvious. 5.32 Comparative Example 7 yes The pungent odor is obvious. 5.35 Comparative Example 8 no Slightly sour aroma 4.42 Comparative Example 9 yes The pungent odor is obvious. 5.45 Comparative Example 10 yes The pungent odor is obvious. 5.37 Comparative Example 11 yes The pungent odor is obvious. 5.53 Comparative Example 12 yes The pungent odor is obvious. 5.49 Comparative Example 13 no Slightly sour aroma 4.47 Comparative Example 14 yes The pungent odor is obvious. 5.97 The feeds obtained in Examples 1-4 and Comparative Examples 1, 2, 3, 8 and 13 were divided into 9 groups, with 5 quails fed to each group. The total daily feed amount was 500g. During the feeding period, free access to feed and water was provided. The total weight of the 5 quails before feeding (on an empty stomach) was recorded as W. 饲喂前总重 Feeding times were 8:00 AM, 2:00 PM, and 6:00 PM for 30 consecutive days. After the last feeding, the five animals were weighed on an empty stomach at 8:00 AM the following day, and the total weight was recorded as W. 饲喂后总重 Calculate the average daily weight gain per animal = (W 饲喂后总重 -W 饲喂前总重 The total number of eggs laid in 30 days was recorded as 5×30. The daily egg production rate of each egg was calculated by "total number of eggs laid / (5×30) × 100%". The eggshell strength was measured by an eggshell strength tester, and the average eggshell strength was calculated. The results are shown in Table 2 below.

[0089] Table 1 Feeding performance Source of materials Average daily weight gain per animal (g) Daily egg production rate per bird (%) <![CDATA[Eggshell strength (kg / cm 2 ).]]> Example 1 2.65 76.67 1.35 Example 2 2.72 83.33 1.36 Example 3 2.85 86.67 1.38 Example 4 2.93 90.00 1.39 Comparative Example 1 1.21 43.33 0.96 Comparative Example 2 1.15 43.33 0.96 Comparative Example 3 1.12 36.67 0.95 Comparative Example 8 1.52 46.67 0.99 Comparative Example 13 1.42 46.67 0.98 As can be seen from Tables 1 and 2 above, the green antibiotic-free quail feed prepared by the present invention has a good odor and low acidity, which not only helps to inhibit the growth of harmful bacteria, but also increases feed intake, which is beneficial to the growth and development of quail, and results in good feeding performance.

[0090] The embodiments described above provide a detailed explanation of the technical solutions and beneficial effects of the present invention. It should be understood that the above descriptions are merely specific embodiments of the present invention and are not intended to limit the present invention. Various changes and modifications can be made to the present invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the present invention as claimed.

Claims

1. A method for preparing a growth-promoting, green, antibiotic-free quail feed, characterized in that, The preparation method includes the following steps: The fermentation substrate is obtained by mixing Chinese herbal medicine residue and water at a weight ratio of 1:0.8~1.4 and then sterilizing at 121℃ for 30 minutes. The fermentation substrate, mixed bacterial strains, and mixed enzymes were mixed in a weight ratio of 1:0.06~0.08:0.008~0.009 and then fermented at 30℃~35℃ for 5~7 days to obtain the green antibiotic-free quail feed.

2. The method for preparing a growth-promoting, green, antibiotic-free quail feed according to claim 1, characterized in that, The mixed strain is OD 600 The Lactobacillus acidophilus bacterial solution had an OD of 0.7-0.

8. 600 The concentrations of domesticated Bacillus subtilis bacterial suspension and OD were 0.6–0.

7. 600 The green wood enzyme culture solution with a concentration of 0.6~0.7 was composed of a weight ratio of 1:0.5~0.6:0.3~0.

5.

3. The method for preparing a growth-promoting, green, antibiotic-free quail feed according to claim 2, characterized in that, The method for preparing the domesticated Bacillus subtilis bacterial solution includes the following steps: The first acclimatized bacterial solution was obtained by culturing Bacillus subtilis microcapsules in a nutrient broth liquid medium with a pH of 5.0 at 28℃~30℃ for 28h~30h. The first acclimatized bacterial solution was cultured in a nutrient broth liquid medium with a pH of 4.0 at 33℃~35℃ for 24h~26h to obtain the second acclimatized bacterial solution; The second acclimated bacteria liquid and the synthetic liquid medium with pH of 6.0 were cultured at 35°C to OD 600 The acclimated Bacillus subtilis liquid was obtained with OD of 0.6-0.

7.

4. The method for preparing a growth-promoting, green, antibiotic-free quail feed according to claim 3, characterized in that, The preparation method of the Bacillus subtilis microcapsules includes the following steps: OD 600 A suspension dispersion was obtained by mixing Bacillus subtilis bacterial suspension (0.7-0.8), oxidized oligosaccharide aqueous dispersion, and amino polysaccharide aqueous dispersion in a weight ratio of 1:18-20:4-5 and reacting at 25°C for 12-18 hours. The suspension and sodium borohydride were mixed at a weight ratio of 1:0.01~0.02 and reacted at 20℃ for 50min~60min. After filtration and washing, the Bacillus subtilis microcapsules were obtained.

5. The method for preparing a growth-promoting, green, antibiotic-free quail feed according to claim 4, characterized in that, The preparation method of the oxidized oligosaccharide aqueous dispersion includes the following steps: Oligosaccharides, sodium periodate, and deionized water were mixed in a weight ratio of 1:0.1~0.2:10~15 and reacted in the dark at 25°C for 5~7 hours. The mixture was then quenched, dialyzed, and freeze-dried to obtain oxidized oligosaccharides. The oxidized oligosaccharide and deionized water were mixed and dispersed at a weight ratio of 0.1:1~2 to obtain the oxidized oligosaccharide aqueous dispersion.

6. The method for preparing a growth-promoting, green, antibiotic-free quail feed according to claim 5, characterized in that, The oligosaccharides include fructooligosaccharides.

7. The method for preparing a growth-promoting, green, antibiotic-free quail feed according to claim 4, characterized in that, The aminopolysaccharide aqueous dispersion includes a chitosan aqueous dispersion with a mass percentage concentration of 1% to 1.6%.

8. The method for preparing a growth-promoting, green, antibiotic-free quail feed according to claim 3, characterized in that, The synthetic liquid culture medium is prepared by mixing 8g-10g glucose, 5g-8g sucrose, 10g-12g peptone, 3g-5g yeast extract, 0.5g-1g L-proline, 0.2g magnesium sulfate heptahydrate and 0.05g manganese sulfate monohydrate, bringing the volume to 1L, and then sterilizing at 121℃ for 20min.

9. The method for preparing a growth-promoting, green, antibiotic-free quail feed according to claim 1, characterized in that, The mixed enzyme consists of cellulase, xylanase and pectinase in a weight ratio of 1:0.2~0.3:0.6~0.

8.

10. A green, non-antibiotic, growth promoting quail feed, characterized in that, The green antibiotic-free quail feed is prepared by any one of the methods described in claims 1 to 9 for preparing a growth-promoting green antibiotic-free quail feed.