Whole-stage rabbit feed based on golden probiotic elements and preparation method thereof

Abstract

The invention relates to the field of feeds, in particular to a full-stage rabbit feed based on golden probiotic elements and a preparation method of the full-stage rabbit feed. The feed comprises a coarse material, prebiotics and probiotics, the prebiotics comprise 5-15 g / kg of fructooligosaccharide and 10-25 g / kg of chicory root powder on the basis of each kg of the whole-stage rabbit feed; the probiotics comprise 1 * 10 < 9 >-3 * 10 < 9 > CFU / Kg of bacillus subtilis, 0.8 * 10 < 9 >-1 * 10 < 9 > CFU / Kg of bacillus licheniformis and 1 * 10 < 9 >-3 * 10 < 9 > CFU / Kg of inactivated lactobacillus acidophilus. All the raw materials have a synergistic effect and are matched with a peptidoglycan enriched fermented soybean meal carrier (the mass ratio is 1: 2), so that (1) active bacterium digestion and mucous membrane repair and inactivated bacterium immune activation are realized; (2) the double-chain prebiotics cover near and far ends of intestinal tracts; and (3) the retention rate of the carrier protected peptidoglycan is greater than or equal to 94.7%, and the problem of high-temperature inactivation is solved.

Description

Technical Field

[0001] This invention relates to the field of feed, and more specifically, to a full-stage rabbit feed based on golden probiotics and its preparation method. Background Technology

[0002] Most commercially available probiotic rabbit food claims to regulate intestinal function, such as alleviating soft stools or promoting digestion, to meet the needs of pet rabbit intestinal health management. These products mainly achieve basic intestinal regulation by adding prebiotics (such as oligosaccharides FOS) or single probiotics (such as Bacillus subtilis), and their mechanism of action is relatively simple. However, professional testing by nutritionists and analysis of extensive user feedback have revealed that while these products do have some intestinal regulatory effects, their formulation design has the following key flaws that limit their overall effectiveness.

[0003] First, the prebiotic coverage is incomplete: these products often use only a single type of short-chain prebiotic (such as FOS) or long-chain prebiotic (such as inulin), failing to achieve synergistic effects. Short-chain prebiotics mainly activate the proximal gut microbiota, while long-chain prebiotics target the distal gut microbiota. However, because they are not used in combination, they cannot simultaneously activate the entire gut microbiota, resulting in regulatory blind spots, such as insufficient proliferation of distal lactobacilli, affecting the overall balance and function of the gut.

[0004] Secondly, probiotics have limited functionality: existing products rely excessively on the digestive functions of live probiotics, such as promoting nutrient absorption through enzymatic fiber breakdown, but lack diverse synergistic mechanisms. For example, the peptidoglycans of inactivated bacteria can stimulate the immune system and enhance immunity in rabbits, while biofilm formation helps block toxins and repair the intestinal mucosa. These key mechanisms are generally missing in the formulations, thus limiting the product's role in immune activation and mucosal repair.

[0005] In addition, high-temperature processing leads to the inactivation of probiotics: during the granulation process, the high-temperature stage (temperature ≥85℃) significantly reduces the survival rate of active probiotics, generally below 60%, and existing production technologies do not employ effective cell protection processes, such as microencapsulation or low-temperature treatment, which further significantly reduces the actual efficacy and stability of the product.

[0006] In summary, current market products cannot systematically address common complex intestinal problems in adult rabbits, such as soft stools, bloating, stress from changing diets, and weakened immunity. Nor can they comprehensively optimize intestinal health to meet the long-term care needs of adult rabbits. Summary of the Invention

[0007] The purpose of this invention is to provide a full-stage rabbit feed based on golden probiotics, which incorporates active probiotics (Bacillus subtilis + Bacillus licheniformis) + inactivated probiotics (Lactobacillus acidophilus) and short-chain prebiotics (FOS) + long-chain prebiotics (inulin). The ingredients work synergistically to achieve multiple effects such as active bacteria digestion, mucosal repair, and inactivated bacteria immune activation. It is suitable for young rabbits, adult rabbits, and old rabbits, and has good compatibility.

[0008] Another objective of this invention is to provide a method for preparing rabbit feed based on golden probiotics for all stages, which can protect the effective components in the feed while effectively killing pathogenic microorganisms and preventing nutrient degradation or mold growth.

[0009] The technical problem solved by this invention is achieved by the following technical solution.

[0010] On the one hand, embodiments of the present invention provide a full-stage rabbit feed based on golden probiotics, including roughage, prebiotics and probiotics; The prebiotics, calculated per kilogram of rabbit feed for all stages, include 5–15 g / kg of fructooligosaccharides and 10–25 g / kg of chicory root powder; The probiotics include Bacillus subtilis 1×10 9 -3×10 9 CFU / Kg, Bacillus licheniformis 0.8×10 9 -1×10 9 CFU / kg, inactivated Lactobacillus acidophilus 1×10 9 –3×10 9 CFU / Kg.

[0011] In some embodiments of the present invention, the prebiotics comprise, per kilogram of rabbit feed for all stages, 6 grams of fructooligosaccharides and 18 grams of chicory root powder. The probiotics include Bacillus subtilis: 1.5 × 10⁻⁶ 9 CFU / Kg, Bacillus licheniformis: 1.0×10 9 CFU / Kg, inactivated Lactobacillus acidophilus: 1.2-1.8×10 9 CFU / Kg.

[0012] In some embodiments of the present invention, the roughage, by weight percentage, includes 35% timothy grass, 11% oat hulls, 10% soybean hulls, 10% barley, 8% oats, 2% beet meal, 5% wheat bran, 6% oat grass, 6% fermented soybean meal, 1.5% vegetable oil, 1.2% yeast powder, 0.7% vitamins, 3.3% minerals, and 0.3% sodium chloride.

[0013] In this embodiment of the invention, the effects of each ingredient are as follows: Fructose oligosaccharides (FOS): A fast-acting biogen. It ferments quickly, rapidly stimulating the proliferation of lactic acid bacteria and bifidobacteria, rapidly lowering intestinal pH, and inhibiting pathogens. It is key to improving diarrhea and soft stools during food transitions.

[0014] Chicory root powder (rich in inulin): long-lasting + physical prebiotic. In addition to its prebiotic properties, its ultra-finely ground fiber provides excellent physical stimulation, promoting intestinal peristalsis. It complements the fermentation rate of fructooligosaccharides, maintaining gut microbiota stability and stool formation. This combination addresses the needs of different stages and microbiota within the gut.

[0015] Bacillus subtilis and Bacillus licheniformis: They are active in the stomach and small intestine, consuming oxygen and creating an anaerobic environment that is conducive to the colonization of anaerobic probiotics in the hindgut.

[0016] It produces enzymes to aid digestion: it secretes proteases, amylases, and non-starch polysaccharide enzymes, which directly improve the digestibility of nutrients (increasing nutrient absorption by 10-15%).

[0017] Biological oxygen depletion and antibacterial activity: Competitively inhibits pathogens such as Escherichia coli and Salmonella, reducing the risk of bloating and diarrhea.

[0018] Inactivated Lactobacillus acidophilus cell fragments (such as peptidoglycan and cell wall components) can act as immunostimulants, significantly enhancing intestinal mucosal and systemic immunity (IgA) and strengthening intestinal barrier function.

[0019] High safety: No competition or risk of surviving bacteria; resistant to processing, storage, and antibiotics; stable in action.

[0020] Timothy hay and oat hulls: provide high-quality long fiber (structural fiber), which is essential for maintaining digestive emptying speed, preventing hairballs, and maintaining fecal pellet structure.

[0021] Soybean hulls, beet pulp, and wheat bran: provide fermentable fiber that is partially utilized by microorganisms in the hindgut to produce volatile fatty acids (providing extra energy for rabbits) and contribute to cecal health.

[0022] Barley and oats: mainly provide starch (which has a high degree of gelatinization after puffing and is easily digestible) and some protein.

[0023] Fermented soybean meal (inactivated bacteria carrier): Provides high-quality protein, and the fermentation process reduces anti-nutritional factors, making it easier to digest. It also serves as a carrier for inactivated bacteria, facilitating uniform mixing and drying.

[0024] Vegetable oils: provide high concentrations of essential fatty acids (such as linoleic acid) and are also carriers of fat-soluble vitamins. Spraying helps reduce dust, improve palatability, and enhance particle gloss.

[0025] Brewer's yeast powder: Rich in B vitamins, nucleic acids, and unknown growth factors, it is a natural palatability enhancer and nutritional supplement, and can also mildly support gut health.

[0026] Vitamin premix, mineral premix, and sodium chloride: ensure comprehensive and balanced nutrition in the feed, meet all micronutrient requirements of adult rabbits, and prevent deficiencies.

[0027] In some embodiments of the present invention, the probiotics further include a carrier, wherein the carrier is fermented soybean meal.

[0028] On the other hand, embodiments of the present invention provide a method for preparing a full-stage rabbit feed based on golden probiotics, comprising the following steps: Preparation of S1 spore powder: Bacillus subtilis and Bacillus licheniformis were inoculated into LB medium containing 0.1% MnCl2 and cultured at 37℃ for 24 h. The culture solution was centrifuged at 8000 rpm for 15 min and spray-dried at 30-45℃ to obtain spore powder. Preparation of S2 inactivated bacteria: Lactobacillus acidophilus was activated in two stages using MRS medium (stage 1: anaerobic at 37℃ for 18 h; stage 2: anaerobic at 37℃ for 12 h with 5% inoculum) to obtain seed culture. The seed culture was then inoculated into a 500L fermenter (containing MRS medium, pH 6.2±0.1) at an inoculum of 8-10%, maintained at 37℃±0.5℃ and dissolved oxygen ≤0.1 ppm (N2 filling). During fermentation, 5M NaOH was automatically added to control the pH at 6.0-6.2. Fermentation was terminated when the residual glucose was ≤1.0 g / L (fermentation time 12-14 h), at which point the viable cell count was ≥2.0 × 10⁻⁶. 10 CFU / mL, peptidoglycan content ≥120mg / L. The fermentation broth was filtered through a 0.22μm microfiltration membrane to remove impurities before entering the inactivation process. The Lactobacillus acidophilus fermentation broth underwent thermal inactivation, centrifuged at 10,000 rpm for 15 min, and concentrated to obtain bacterial sludge. This sludge was mixed with fermented soybean meal at a mass ratio of 1:2 and air-dried at 50-55℃ until the moisture content was ≤8%, thus obtaining inactivated bacteria. Preparation of S3 chicory root powder: Chicory root powder is obtained by ultra-fine grinding of dried chicory roots to a particle size ≤180μm (80 mesh). S4 gradient blending: Mix timothy grass, oat hulls, and soybean hulls dry at 25 rpm for 5 minutes; then add barley, oats, beet pulp, and wheat bran and mix for 3 minutes. Finally, add Bacillus thuringiensis powder, inactivated bacteria, chicory root powder, and fructooligosaccharides and mix at 30 rpm for 10 minutes. S5 oils and trace components blending: Vegetable oil was sprayed into the mixture from step S4, and vitamin premix, mineral premix, sodium chloride, and brewer's yeast powder were added. The mixture was then mixed until the coefficient of variation (CV) was ≤5%. S6 High Temperature Tempering and Expansion Molding: In the conditioner, the material was conditioned at a steam pressure of 0.35 MPa, a moisture content of 18.0 ± 0.5%, a temperature of 85 ± 2℃, and a residence time of 90 s. In a twin-screw extruder, the temperature is 98±2℃ and the barrel pressure is 18. At 22 MPa and a residence time ≤30 s, the conditioned material is expanded to obtain columnar granules. S7 Cooling: In a fluidized bed, the pellets are cooled for 5 minutes at a cold air temperature of 25°C and a wind speed of 2.5 m / s until the internal temperature of the columnar particles is ≤30°C; then vibrating and sieving are performed to obtain the rabbit feed for all stages.

[0029] In some embodiments of the present invention, the vegetable oil is added at a rate of 15 g / kg per kilogram of rabbit feed for all stages. In some embodiments of the present invention, the vegetable oil is at least one of rapeseed oil, peanut oil, and sunflower seed oil. Compared with the prior art, the embodiments of the present invention have at least the following advantages or beneficial effects: The rabbit feed provided by this invention incorporates active probiotics (Bacillus subtilis + Bacillus licheniformis) + inactivated probiotics (Lactobacillus acidophilus) and short-chain prebiotics (FOS) + long-chain prebiotics (inulin) into the roughage. The synergistic effect of these ingredients, combined with a peptidoglycan-enriched fermented soybean meal carrier (mass ratio 1:2), achieves a triple effect: (1) Active bacteria digestion and mucosal repair + inactivated bacteria immune activation; (2) Double-chain prebiotics cover the proximal and distal parts of the intestine; (3) The carrier protects the peptidoglycan retention rate ≥94.7%, solving the problem of high temperature inactivation.

[0030] Carrier protection mechanism: The fermented soybean meal carrier achieves high-temperature protection of peptidoglycan through a triple synergistic effect: (1) Physical barrier: The 3-50μm microporous structure formed by fermentation spatially confines the bacterial mud, blocking direct heat conduction; (2) Molecular anchoring: β-glucan in the carrier forms a hydrogen bond network with the acetyl muramic acid of peptidoglycan, and phosphate groups and calcium ions construct ionic crosslinks; (3) Thermal buffering effect: The water holding capacity of the carrier is ≥4g water / g protein, and the heat absorption of water evaporation makes the actual heating temperature of peptidoglycan ≤68℃ (below the denaturation threshold). The triple mechanism makes the peptidoglycan retention rate ≥94.7% under the 98℃ / 30s puffing condition, which is significantly higher than that of conventional carriers (wheat bran ≤76.5%). Solid fermented soybean meal is used as the carrier for inactivating bacteria. Its β-glucanase activity is ≥500U / g. The bacterial mud and fermented soybean meal are loaded at a ratio of 1:2. Its peptidoglycan retention rate is ≥94.7%, which breaks through the limitations of active bacteria and uses peptidoglycan to achieve long-term immune stimulation.

[0031] The preparation method provided by this invention adds MnCl2 to promote spore formation, improve bacterial tolerance and colonization ability, and help maintain the balance of intestinal flora. Inactivated bacteria have immunomodulatory functions, can enhance the intestinal barrier, and are highly stable, easy to store and process. Chicory root powder (ultra-finely ground): rich in inulin and other prebiotics, it works synergistically with probiotics to exert a dual regulatory effect of "bacteria + prebiotics," promoting intestinal health. The proportions of crude fiber sources such as timothy grass, oat hulls, and soybean hulls are reasonable, meeting the physiological needs of rabbits' herbivorous diet and helping to maintain digestive tract motility and dental health.

[0032] Step-by-step feeding and speed control ensure uniform distribution of trace components (vitamins, minerals, probiotics, etc.), avoiding gradation and guaranteeing product uniformity. Conditioning parameters of 85℃, 18% moisture content, and 90s allow for thorough starch gelatinization, improving digestibility while simultaneously killing some pathogenic microorganisms. High-temperature, high-pressure, and instantaneous processing at 98℃, 18–22 MPa, and ≤30s further sterilizes and improves starch gelatinization, creating a porous structure conducive to digestion; short-time processing reduces the loss of heat-sensitive nutrients. Low-temperature spray drying and airflow drying preserve bacterial activity in the spore powder (30–45℃); inactivated bacterial airflow drying (50–55℃) avoids excessive heat damage and preserves bacterial function. Fluidized bed cold air cooling rapidly reduces particle temperature to ≤30℃, preventing residual heat from causing nutrient degradation or mold growth. Detailed Implementation

[0033] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0034] It should be noted that, unless otherwise specified, the embodiments and features described in the present invention can be combined with each other. The present invention will now be described in detail with reference to specific embodiments.

[0035] Example 1 Prepare the ingredients according to the following proportions: The prebiotics, per kilogram of rabbit feed for all stages, include 6 grams of fructooligosaccharides and 18 grams of chicory root powder. The probiotics include Bacillus subtilis: 1.5 × 10⁻⁶ 9 CFU / Kg, Bacillus licheniformis: 1.0×10 9 CFU / Kg, inactivated Lactobacillus acidophilus: 1.2×10 9 CFU / Kg.

[0036] By weight percentage, the roughage includes 35% timothy hay, 11% oat hulls, 10% soybean hulls, 10% barley, 8% oats, 2% beet meal, 5% wheat bran, 6% oat grass, 6% fermented soybean meal, 1.5% vegetable oil, 1.2% yeast powder, 0.7% vitamin premix, 3.3% mineral premix, and 0.3% sodium chloride.

[0037] The vitamin premix includes vitamin C, vitamin E, vitamin A, vitamin D3, vitamin K3, vitamin B1, vitamin B2, vitamin B6, vitamin B12, niacin, folic acid, D-calcium pantothenate, D-biotin, and choline chloride.

[0038] The mineral premix includes calcium carbonate, dicalcium phosphate, calcium dihydrogen phosphate, sodium chloride, glycine copper chelate, glycine iron chelate, methionine zinc chelate, methionine manganese chelate, calcium iodate, yeast selenium, and yeast chromium.

[0039] Prepare rabbit feed for all stages using the following steps: Preparation of S1 spore powder: Bacillus subtilis and Bacillus licheniformis were inoculated into LB medium containing 0.1% MnCl2 and cultured at 37℃ for 24 h. The culture solution was centrifuged at 8000 rpm for 15 min and spray-dried at 30-45℃ to obtain spore powder. Preparation of S2 inactivated bacteria: Lactobacillus acidophilus (accession number CICC 6078) was activated in two stages on MRS medium (stage 1: anaerobic at 37℃ for 18 h; stage 2: anaerobic at 37℃ for 12 h with 5% inoculum) to obtain seed culture. The seed culture was inoculated into a 500L fermenter (containing MRS medium, pH 6.2±0.1) at an inoculum of 8-10%, maintained at 37℃±0.5℃ and dissolved oxygen ≤0.1 ppm (N2 filling). During fermentation, 5M NaOH was automatically added to control the pH at 6.0-6.2. Fermentation was terminated when the residual glucose was ≤1.0 g / L (fermentation time 12-14 h), at which point the viable cell count was ≥2.0 × 10⁻⁶. 10 CFU / mL, peptidoglycan content ≥120mg / L. The fermentation broth was filtered through a 0.22μm microfiltration membrane to remove impurities before entering the inactivation process. The Lactobacillus acidophilus fermentation broth underwent thermal inactivation, centrifuged at 10000 rpm for 15 min, and concentrated to obtain bacterial sludge. This sludge was mixed with fermented soybean meal at a mass ratio of 1:2 and air-dried at 50-55℃ until the moisture content was ≤8%, thus obtaining inactivated bacteria. Preparation of S3 chicory root powder: Chicory root powder is obtained by ultra-fine grinding of dried chicory roots to a particle size ≤180μm (80 mesh). S4 gradient blending: Mix timothy grass, oat hulls, and soybean hulls dry at 25 rpm for 5 minutes; then add barley, oats, beet pulp, and wheat bran and mix for 3 minutes. Finally, add Bacillus thuringiensis powder, inactivated bacteria, chicory root powder, and fructooligosaccharides and mix at 30 rpm for 10 minutes. S5 oils and trace components blending: Vegetable oil (rapeseed oil) is sprayed into the mixture in step S4, and vitamin premix, mineral premix, sodium chloride, and brewer's yeast powder are added. The mixture is then mixed until the coefficient of variation (CV) is ≤5%. S6 High Temperature Tempering and Expansion Molding: In the conditioner, the material was conditioned at a steam pressure of 0.35 MPa, a moisture content of 18.0 ± 0.5%, a temperature of 85 ± 2℃, and a residence time of 90 s. In a twin-screw extruder, the temperature is 98±2℃ and the barrel pressure is 18. At 22 MPa and a residence time ≤30 s, the conditioned material is expanded to obtain columnar granules. S7 Cooling: In a fluidized bed, the pellets are cooled for 5 minutes at a cold air temperature of 25°C and a wind speed of 2.5 m / s until the internal temperature of the columnar particles is ≤30°C; then vibrating and sieving are performed to obtain the rabbit feed for all stages.

[0040] Example 2 Prepare the ingredients according to the following proportions: The prebiotics, per kilogram of rabbit feed for all stages, include 5 grams of fructooligosaccharides and 25 grams of chicory root powder. The probiotics include Bacillus subtilis: 3 × 10 9 CFU / Kg, Bacillus licheniformis: 0.8×10 9 CFU / Kg, inactivated Lactobacillus acidophilus: 1.8×10 9 CFU / Kg.

[0041] By weight percentage, the roughage comprises 35% timothy hay, 11% oat hulls, 10% soybean hulls, 10% barley, 8% oats, 2% beet meal, 5% wheat bran, 6% oat grass, 6% fermented soybean meal, 1.5% vegetable oil, 1.0% yeast powder, 0.7% vitamin premix, 3.3% mineral premix, and 0.3% sodium chloride. The preparation method for the rabbit feed throughout all stages is the same as in Example 1.

[0042] Example 3 Prepare the ingredients according to the following proportions: The prebiotics, per kilogram of rabbit feed for all stages, include 15 grams of fructooligosaccharides and 10 grams of chicory root powder. The probiotics include Bacillus subtilis: 1×10 9 CFU / Kg, Bacillus licheniformis: 1×10 9 CFU / kg, inactivated Lactobacillus acidophilus: 1×10 9 CFU / Kg.

[0043] By weight percentage, the roughage comprises 35% timothy hay, 11% oat hulls, 10% soybean hulls, 10% barley, 8% oats, 2% beet meal, 5% wheat bran, 6% oat grass, 6% fermented soybean meal, 1.5% vegetable oil, 1.0% yeast powder, 0.7% vitamin premix, 3.3% mineral premix, and 0.3% sodium chloride.

[0044] The preparation method of the rabbit feed for all stages is the same as that in Example 1.

[0045] Example 4 The difference from Example 1 is that, by mass percentage, the proportion of coarse material is: Timothy hay 40%, oat hulls 13%, soybean hulls 12%, barley 8%, oats 5%, beet meal 5%, wheat bran 4%, fermented soybean meal 6%, vegetable oil 1.2%, yeast powder 1.5%, vitamin premix 0.7%, mineral premix 3.3%, sodium chloride 0.3%.

[0046] The remaining raw materials and preparation methods are the same as in Example 1.

[0047] Example 5 The difference from Example 1 is that, by mass percentage, the proportion of coarse material is: Timothy hay 28%, oat hulls 16%, soybean hulls 8%, barley 15%, oats 12%, beet meal 1%, wheat bran 6%, fermented soybean meal 6%, vegetable oil 2.2%, yeast powder 1.8%, vitamin premix 0.7%, mineral premix 3.0%, sodium chloride 0.3%.

[0048] The remaining raw materials and preparation methods are the same as in Example 1.

[0049] Comparative Example 1 The difference from Example 1 is that only fructooligosaccharides and crude materials are added, and the proportions are the same as in Example 1.

[0050] Comparative Example 2 The difference from Example 1 is that only chicory root powder and coarse materials are added, and the proportions are the same as in Example 1.

[0051] Comparative Example 3 The difference from Example 1 is that only prebiotics and coarse feed are added, and the proportions are the same as in Example 1.

[0052] Experimental Example 1. Laboratory animals and grouping Experimental animals: Select healthy adult New Zealand rabbits or Ira rabbits of similar weight (e.g., 3.0-3.5 kg), and set up 10 groups.

[0053] Example group: fed the feed of Examples 1-5; Comparative control group: fed with the diet of comparative control groups 1-3; Control group: CK-B: Feed a basic diet (containing only roughage, vitamins, minerals, etc., without any functional additives). CK-C: Feeding rabbits with mainstream commercially available rabbit feed for all stages (as a market comparison). 2. Experimental period and feeding management Pre-feeding period: 7 days. All rabbits are fed a basic diet to allow them to adapt to the environment.

[0054] Formal experimental period: 28 days. Each group was fed the corresponding feed.

[0055] Food transition period: The first 7 days of the formal experimental period are considered the food transition period, which is used to observe diarrhea.

[0056] Management: Free access to food and water, standard rabbit hutch environment, and regular disinfection.

[0057] 3. Measurement Indicators and Methods Growth performance and health indicators: Soft stool rate: Observe and record the number of rabbits with soft stool (poorly formed, high water content) daily. Soft stool rate (%) = (Number of rabbits with soft stool / Total number of rabbits in the group) × 100%.

[0058] Incidence of bloating: Rabbits were observed and their abdomens were distended and tympanic upon percussion were recorded daily. Incidence of bloating (%) = (Number of rabbits with bloating / Total number of rabbits in the group) × 100%.

[0059] Diarrhea rate during food transition: Record the number of rabbits that develop watery or pasty diarrhea during the first 7 days of food transition.

[0060] Digestive and immune indicators: Crude fiber digestibility: A total fecal collection method experiment was conducted during the last 5 days of the experiment. Crude fiber digestibility (%) = [(ingested crude fiber - amount of crude fiber in feces) / ingested crude fiber] × 100%.

[0061] Serum IgA level: At the end of the experiment, a certain number of rabbits were randomly selected from each group, blood was collected from the anterior vena cava, serum was separated, and IgA concentration was measured using an ELISA kit.

[0062] Apparent digestibility of nutrients (dry matter / protein): Same as fecal collection method. Apparent digestibility (%) = [(mass of ingested nutrients - mass of nutrients in feces) / mass of ingested nutrients] × 100%.

[0063] Stool morphology scoring: A 1-5 point scale was used (1 point: watery; 2 points: soft stool; 3 points: well formed; 4 points: dry and formed; 5 points: dry and hard pellets). The scores were calculated daily and the average value was taken.

[0064] The results are shown in Table 1.

[0065] Table 1

[0066] As shown in Table 1, the rabbit feed provided in the examples reduces the rate of soft stools by 50%, the incidence of bloating by 60%, increases the digestibility of crude fiber by 30% (due to dried and formed feces), reduces the diarrhea rate during the feed transition period by 50%, increases serum IgA antibody levels by 35%, and improves the measured nutrient absorption rate by 10–15%.

[0067] In summary, the embodiments of this invention incorporate active probiotics (Bacillus subtilis + Bacillus licheniformis) + inactivated probiotics (Lactobacillus acidophilus) and short-chain prebiotics (FOS) + long-chain prebiotics (inulin) into the roughage. The synergistic effect of these ingredients, combined with a peptidoglycan-enriched fermented soybean meal carrier (mass ratio 1:2), achieves a synergistic effect of active bacterial digestion, mucosal repair, and inactivated bacterial immune activation. The double-chain prebiotics cover the proximal and distal ends of the intestine; the carrier protects the peptidoglycan retention rate at ≥94.7%, solving the problem of high-temperature inactivation. The rabbit feed provided in these embodiments is suitable for young, adult, and old rabbits, covering all stages, with good adaptability, and has effects such as mucosal repair and inactivated bacterial immune activation in rabbits of different ages.

[0068] Solid-state fermented soybean meal was used as a carrier for inactivated bacteria, with β-glucanase activity ≥500U / g. The bacterial sludge and fermented soybean meal were loaded at a ratio of 1:2, and the peptidoglycan retention rate was ≥94.7%, which broke through the limitations of live bacteria and achieved long-term immune stimulation using peptidoglycan.

[0069] The embodiments described above are some, but not all, embodiments of the present invention. The detailed description of the embodiments of the present invention is not intended to limit the scope of the claimed invention, but merely to illustrate selected embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

Claims

1. A rabbit feed for all stages based on golden probiotics, characterized in that, Including roughage, prebiotics, and probiotics; The prebiotics, calculated per kilogram of rabbit feed for all stages, include 5–15 g / kg of fructooligosaccharides and 10–25 g / kg of chicory root powder; The probiotics include Bacillus subtilis 1×10 9 -3×10 9 CFU / Kg, Bacillus licheniformis 0.8×10 9 -1×10 9 CFU / kg, inactivated Lactobacillus acidophilus 1×10 9 –3×10 9 CFU / Kg.

2. The rabbit feed based on golden probiotics for all stages according to claim 1, characterized in that, The prebiotics, calculated per kilogram of rabbit feed for all stages, include 6 grams of fructooligosaccharides and 18 grams of chicory root powder. The probiotics include Bacillus subtilis: 1.5 × 10⁻⁶ 9 CFU / Kg, Bacillus licheniformis: 1.0×10 9 CFU / Kg, inactivated Lactobacillus acidophilus: 1.2-1.8×10 9 CFU / Kg.

3. The rabbit feed based on golden probiotics for all stages according to claim 1, characterized in that, By weight percentage, the coarse feed comprises 35% timothy grass, 11% oat hulls, 10% soybean hulls, 10% barley, 8% oats, 2% beet meal, 5% wheat bran, 6% oat grass, 6% fermented soybean meal, 1.5% vegetable oil, 1.2% yeast powder, 0.7% vitamins, 3.3% minerals, and 0.3% sodium chloride.

4. A method for preparing a full-stage rabbit feed based on golden probiotics as described in any one of claims 1-3, characterized in that, Includes the following steps: Preparation of S1 spore powder: Bacillus subtilis and Bacillus licheniformis were inoculated into LB medium containing 0.1% MnCl2 and cultured at 37℃ for 24 h. The culture solution was centrifuged at 8000 rpm for 15 min and spray-dried at 30-45℃ to obtain spore powder. Preparation of S2 inactivated bacteria: Lactobacillus acidophilus was activated twice on MRS medium, then fermented under the conditions of MRS medium, pH 6.2±0.1, maintained at 37℃±0.5℃, and dissolved oxygen ≤0.1 ppm. After filtration to remove impurities, a viable count ≥2.0×10⁻⁶ was obtained. 10 Lactobacillus acidophilus fermentation broth with CFU / mL and peptidoglycan content ≥120mg / L; The fermentation broth of Lactobacillus acidophilus was thermally inactivated, centrifuged at 10,000 rpm for 15 min, and concentrated to obtain bacterial sludge. The bacterial sludge was mixed with fermented soybean meal at a mass ratio of 1:2 and dried in an airflow at 50-55℃ until the moisture content was ≤8%, thus obtaining inactivated bacteria. Preparation of S3 chicory root powder: Chicory root powder is obtained by ultra-fine grinding of dried chicory roots to a particle size ≤180μm (80 mesh). S4 gradient blending: Mix timothy grass, oat hulls, and soybean hulls dry at 25 rpm for 5 minutes; then add barley, oats, beet pulp, and wheat bran and mix for 3 minutes. Finally, add Bacillus thuringiensis powder, inactivated bacteria, chicory root powder, and fructooligosaccharides and mix at 30 rpm for 10 minutes. S5 oils and trace components blending: Add vegetable oil to the mixture from step S4, and then add vitamin premix, mineral premix, sodium chloride, and brewer's yeast powder, mixing until the coefficient of variation (CV) is ≤5%. S6 High Temperature Tempering and Expansion Molding: In the conditioner, the material was conditioned at a steam pressure of 0.35 MPa, a moisture content of 18.0 ± 0.5%, a temperature of 85 ± 2℃, and a residence time of 90 s. In a twin-screw extruder, the temperature is 98±2℃ and the barrel pressure is 18. At 22 MPa and a residence time ≤30 s, the conditioned material is expanded to obtain columnar granules. S7 Cooling: In a fluidized bed, the pellets are cooled for 5 minutes at a cold air temperature of 25°C and a wind speed of 2.5 m / s until the internal temperature of the columnar particles is ≤30°C; then vibrating and sieving are performed to obtain the rabbit feed for all stages.

5. The method for preparing rabbit feed based on golden probiotics for all stages according to claim 4, characterized in that, The amount of vegetable oil added is 15g / kg per kilogram of rabbit feed for all stages.

6. The method for preparing rabbit feed based on golden probiotics for all stages according to claim 4, characterized in that, The vegetable oil is at least one of rapeseed oil, peanut oil, and sunflower seed oil.

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