A synergistic additive of complex probiotics and plant extracts for antibiotic-free farming

Abstract

The application discloses a kind of complex probiotics and plant extract synergistic additive for no-antibiotic breeding, and relates to the technical field of animal feed additives.The present application comprises complex probiotics, synergistic plant extract, synergistic carrier and stabilizing protective agent.The present application realizes the physical isolation of probiotics and plant extract by intercalation of nano montmorillonite and extract of goldthread, eucommia ulmoides, rosemary and cinnamon and microcapsule embedding, to avoid direct contact inhibition;Through double-layer microcapsule and low-temperature blending process, the sealed survival rate can be greatly improved, and it has high stability;Nano-emulsification improves the bioavailability of plant extract and reduces the direct contact inhibition of probiotics;Double-wall material microcapsule protects probiotics from passing through gastric acid environment, and releases low-temperature blending in the intestinal target to avoid activity loss, and the product storage stability is better than existing preparations.

Description

Technical Field

[0001] This invention relates to the field of animal feed additives, specifically to a synergistic additive of antibiotic-free probiotics and plant extracts for animal husbandry. Background Technology

[0002] Probiotics are live microorganisms that, when given a certain number, can produce beneficial effects on the health of the host. They mainly include Bifidobacterium, Lactobacillus, Clostridium butyricum, Saccharomyces boulardii, Enterococcus, Bacillus licheniformis, and Bacillus cereus.

[0003] In the prior art, probiotics and plant extracts are two major types of antibiotic alternatives that have been widely used. However, most existing probiotic additives use single strains or simple mixed strains, which cannot fully replace the multiple effects of antibiotics. Although existing plant extracts have antibacterial and anti-inflammatory activities, they only have single functions of inhibiting bacteria or inflammation and cannot replace the dual effects of antibiotics in promoting growth and maintaining intestinal health. Therefore, this invention provides a synergistic additive of compound probiotics and plant extracts for antibiotic-free animal husbandry. Summary of the Invention

[0004] To address the shortcomings of existing technologies, this invention provides a synergistic additive of antibiotic-free probiotics and plant extracts for aquaculture, thereby solving the aforementioned problems.

[0005] To achieve the above objectives, the present invention provides the following technical solution: a synergistic additive of antibiotic-free probiotics and plant extracts for aquaculture, comprising the following raw materials by weight: Compound probiotic powder: 30-50 parts, wherein the compound probiotics include Bifidobacterium animalis subsp. lactis V9, Pediococcus lactis PA-19, Bacillus subtilis BS-2, Clostridium butyricum CB-1 and Saccharomyces cerevisiae SC-22; Synergistic plant extract complex: 20-35 parts, wherein the plant extracts include extracts of *Gnaphalium affine*, *Eucommia ulmoides*, rosemary, and cinnamon; Synergistic carrier: 15-25 parts, including xylooligosaccharides, β-glucan and nano-montmorillonite; Stabilizing and protective agent: 5-10 parts, including trehalose and sodium caseinate.

[0006] Preferably, the weight parts of each component in the compound probiotic powder are as follows: Bifidobacterium animalis subsp. lactis V9: 10–15 portions; Pediococcus lactis PA-19: 8–12 portions; Bacillus subtilis BS-2: 5-10 parts; Clostridium butyricum CB-1: 5-8 parts; Saccharomyces cerevisiae SC-22: 2 to 5 parts.

[0007] Preferably, the weight parts of each component in the synergistic plant extract complex are as follows: Extract of *Cephalotaxus fortunei*: 8–12 parts; Eucommia ulmoides extract: 5-8 parts; Rosemary extract: 4-7 parts; Cinnamon extract: 3-5 parts.

[0008] Preferably, the weight parts of each component in the synergistic carrier are as follows: Xylooligosaccharides: 8–12 parts; β-glucan: 4–7 parts; Nano-montmorillonite: 3-6 parts.

[0009] Preferably, the weight parts of each component in the stabilizing and protective agent are as follows: Trehalose: 3-5 parts; Sodium caseinate: 2-5 parts.

[0010] A method for preparing the additive described in any one of the above claims includes the following steps: A1. Preparation of synergistic plant extract composite carrier: extracts of Angelica sinensis, Eucommia ulmoides, rosemary, and cinnamon are nanoemulsified and intercalated with nano-montmorillonite to form a plant extract composite carrier. A2. Preparation of compound probiotic microcapsules: Compound probiotic powder is encapsulated in microcapsules using trehalose and sodium caseinate as double-layer wall materials, wherein the inner wall material is trehalose and the outer wall material is sodium caseinate, thus obtaining double-layer wall material microcapsules. A3. Low-temperature blending: Under nitrogen protection at 4–10°C, the composite carrier of A1, the microcapsules of A2, xylooligosaccharides, and β-glucan are placed in a three-dimensional mixer for blending for 20–40 minutes to obtain a synergistic additive.

[0011] Preferably, the nanoemulsification described in A1 specifically employs a high-pressure homogenization method, which includes: A11. Mix the extracts of *Polygonum multiflorum*, *Eucommia ulmoides*, *Rosemaryia pubescens*, and *Cinnamomum cassia* in proportion, add an ethanol-water mixture to dissolve them completely, and then add 2-5% of Tween-80 as an emulsifier as the total weight of the plant extracts. Stir magnetically at 35-45°C for 30-45 minutes to form a preliminary emulsion, wherein the volume ratio of ethanol to water is 1:3. A12. Transfer the prepared emulsion to a high-pressure homogenizer, set the homogenization pressure to 80-120MPa, the homogenization temperature to 25-35℃, and continuously cycle the homogenization 3-5 times. Detect the particle size of the emulsion using a laser particle size analyzer until more than 90% of the particles are distributed in the range of 50-100nm, and obtain a nano-emulsified plant extract mixture. A13. The prepared nano-emulsified plant extract mixture is stirred at 45°C for 1.5 hours, then freeze-dried under vacuum and pulverized to a particle size of less than 80nm to form a stable plant extract composite carrier.

[0012] Preferably, the preparation of the double-walled microcapsules described in A2 employs a combined spray condensation-freeze-drying technique, specifically including: A21. Mix the compound probiotic powder with trehalose in a certain proportion, add an appropriate amount of sterile water and stir until completely dissolved to form a uniform core material-inner wall material mixture. A22. The mixture is atomized by a high-pressure spray device at a spray pressure of 0.3 to 0.5 MPa. The atomized droplets are sprayed into a liquid paraffin condensation medium at -15 to -10°C to form probiotic microspheres that encapsulate the inner wall material. The microspheres are collected and washed 2 to 3 times with sterile anhydrous ethanol to remove residual liquid paraffin on the surface, thus obtaining the preliminary encapsulated microsphere intermediate.

[0013] Preferably, the low-temperature blending described in A3 specifically includes: A31. First, premix xylooligosaccharides and β-glucan for 5-10 minutes; A32. Add the plant extract complex carrier and mix for 10-15 minutes. A33. Add the double-layer wall material microcapsules and mix for 5-10 minutes.

[0014] Beneficial effects Compared with the prior art, the present invention has the following advantages: Physical isolation between probiotics and plant extracts is achieved through intercalation of nano-montmorillonite with extracts of *Eucommia ulmoides*, *Rosemaryia pubescens*, and *Cinnamomum cassia*, followed by microencapsulation, thus avoiding direct contact inhibition. The double-layer microcapsule and low-temperature blending process significantly improves the sealed survival rate and exhibits high stability. Nanoemulsification enhances the bioavailability of plant extracts and reduces direct contact inhibition of probiotics. The double-layer wall microcapsule protects probiotics through the acidic gastric environment, enabling targeted release in the intestines. Low-temperature blending prevents activity loss, resulting in product storage stability superior to existing formulations. Attached Figure Description

[0015] Figure 1 This is a flowchart of the process of this invention; Figure 2 This is a flowchart illustrating the process of preparing the synergistic plant extract composite carrier in this invention; Figure 3 This is a flowchart illustrating the process of preparing compound probiotic microcapsules in this invention; Figure 4 This is a flowchart of the low-temperature blending process in this invention. Detailed Implementation

[0016] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. 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.

[0017] This invention provides a synergistic additive of antibiotic-free probiotics and plant extracts for aquaculture, comprising the following raw materials by weight: Compound probiotic powder: 30-50 parts, wherein the compound probiotics include Bifidobacterium animalis subsp. lactis V9, Pediococcus lactis PA-19, Bacillus subtilis BS-2, Clostridium butyricum CB-1 and Saccharomyces cerevisiae SC-22; Synergistic plant extract complex: 20-35 parts, plant extracts including angelica root extract, eucommia bark extract, rosemary extract and cinnamon extract; Synergistic carrier: 15-25 parts, including xylooligosaccharides, β-glucan and nano-montmorillonite; Stabilizing and protective agent: 5-10 parts, including trehalose and sodium caseinate.

[0018] Specifically, the weight percentages of each component in the compound probiotic powder are as follows: Bifidobacterium animalis subsp. lactis V9: 10-15 portions. Bifidobacterium animalis subsp. lactis V9 has strong acid and bile salt resistance and intestinal adhesion ability. Pediococcus lactis PA-19: 8-12 parts; Pediococcus lactis PA-19 can produce bacteriocins to inhibit pathogens. Bacillus subtilis BS-2: 5-10 parts. Bacillus subtilis BS-2 can produce extracellular enzyme systems (protease, amylase, cellulase) to promote feed digestion. Clostridium butyricum CB-1: 5-8 parts. Clostridium butyricum CB-1 can produce butyric acid to repair the intestinal mucosa. Saccharomyces cerevisiae SC-22: 2-5 parts. Saccharomyces cerevisiae SC-22 can provide B vitamins and unknown growth factors. Furthermore, the viable count ratio of Bifidobacterium animalis subsp. lactis V9, Pediococcus lactis PA-19, Bacillus subtilis BS-2, Clostridium butyricum CB-1, and Saccharomyces cerevisiae SC-22 was (3-5):(2-4):(1.5-3):(1.5-2.5):1. This ratio can fully leverage the synergistic and complementary effects among the strains, enabling Bifidobacterium animalis subsp. lactis V9 and Pediococcus lactis PA-19 to rapidly colonize the intestines and form a dominant bacterial community, inhibiting the growth of harmful bacteria; Bacillus subtilis BS-2... The extracellular enzyme system can simultaneously decompose indigestible components in feed, improving nutrient utilization; butyric acid secreted by Clostridium butyricum CB-1 can promptly repair intestinal mucosal damage and enhance intestinal barrier function; B vitamins and growth factors provided by Saccharomyces cerevisiae SC-22 provide necessary nutritional support for the growth and reproduction of other probiotics, enhance the colonization ability and activity of compound probiotics, thereby effectively improving the intestinal microecological balance of animals, enhancing the body's immunity, reducing the need for antibiotic use during the breeding process, and meeting the requirements of antibiotic-free breeding.

[0019] Specifically, the weight parts of each component in the synergistic plant extract complex are as follows: Bo Luo Hui extract: 8-12 parts. Bo Luo Hui extract contains benzophenanthridine alkaloids such as sanguisorbin and chelidonine, which have broad-spectrum antibacterial and anti-inflammatory activities. Eucommia ulmoides extract: 5-8 parts. Eucommia ulmoides extract contains chlorogenic acid and genipin, which have immunomodulatory and antioxidant functions. Rosemary extract: 4-7 parts. Rosemary extract contains carrageenan and rosmarinic acid, which are natural antioxidants that protect the activity of probiotics. Cinnamon extract: 3-5 parts. Cinnamon extract contains cinnamaldehyde, which enhances intestinal blood circulation and nutrient absorption. Furthermore, the extracts contain sanguisorbin ≥40wt%, chlorogenic acid ≥20wt% in Eucommia ulmoides extract, caryophyllic acid ≥30wt% in rosemary extract, and cinnamaldehyde ≥25wt% in cinnamon extract. This configuration fully leverages the synergistic effects among the plant extracts, effectively complementing the compound probiotics. The broad-spectrum antibacterial activity of the extract helps probiotics inhibit harmful intestinal bacteria and reduce pathogen colonization. The natural antioxidant properties of the rosemary extract effectively protect the activity of the compound probiotics during storage and in the animal's intestines, prolonging their duration of action. The immunomodulatory function of the Eucommia ulmoides extract, combined with the effect of probiotics in improving the intestinal microecology, further enhances the animal's immunity. The cinnamon extract promotes intestinal blood circulation, improving the absorption efficiency of probiotics and nutrients. The standard content of the effective components in each extract ensures the stability and reliability of its efficacy. Working together with the compound probiotics, synergistic carriers, and stabilizing agents, it enhances the overall effect of the additive, contributing to the dual improvement of animal health and farming efficiency under antibiotic-free farming.

[0020] Specifically, the weight parts of each component in the synergistic carrier are as follows: Xylooligosaccharides: 8-12 parts; Xylooligosaccharides can selectively promote the proliferation of Bifidobacteria. β-glucan: 4-7 parts. β-glucan can activate macrophages and gut-associated lymphoid tissue. Nano-montmorillonite: 3-6 parts. Nano-montmorillonite can adsorb mycotoxins and act as a carrier for probiotic intestinal colonization. Furthermore, nano-montmorillonite, after activation treatment at 200–300℃, has a specific surface area ≥200m². 2 / g, interlayer spacing ≥1.5nm; This configuration fully leverages the synergistic and complementary effects between carrier components, creating a synergistic effect with the compound probiotics and plant extracts. The selective proliferation of xylooligosaccharides enhances the colonization advantage of Bifidobacteria in the compound probiotics, further consolidating the structure of beneficial intestinal flora. The immune cells activated by β-glucan synergistically enhance the non-specific immune response of the animal. After specific activation treatment, the high specific surface area and suitable interlayer spacing of nano-montmorillonite not only efficiently adsorb mycotoxins in feed and the intestines, reducing the damage of toxins to the animal's intestines and probiotics, but also provide colonization attachment sites for probiotics, promoting their adhesion and colonization on the intestinal mucosa. The synergistic effect of xylooligosaccharides, β-glucan, and nano-montmorillonite effectively improves the stability and efficacy of the compound probiotics, while also increasing the efficiency of the plant extracts. Together with stabilizing agents, it ensures the activity of the additive during storage and application, providing solid support for the intestinal health and growth performance of animals in antibiotic-free farming.

[0021] Specifically, the weight parts of each component in the stabilizing and protective agent are as follows: Trehalose: 3-5 parts. Trehalose can improve the survival rate of freeze-dried probiotics. Sodium caseinate: 2-5 parts, sodium caseinate is used as the microcapsule wall material; Furthermore, the weight ratio of trehalose to sodium caseinate is (1.2-1.5):1. This ratio fully leverages the synergistic protective effects of trehalose and sodium caseinate. Trehalose's molecular-level protection mechanism maintains the integrity of probiotic cell membranes during freeze-drying, enhancing the survival rate of live bacteria. Sodium caseinate, as a microcapsule wall material, forms a dense and sustained-release encapsulation layer, effectively isolating probiotics from external oxygen, humidity, and the erosion of gastric acid and bile salts in the digestive tract.

[0022] Please see Figure 1-4 A method for preparing the above-mentioned additive includes the following steps: A1. Preparation of synergistic plant extract composite carrier: extracts of Angelica sinensis, Eucommia ulmoides, rosemary, and cinnamon are nanoemulsified and intercalated with nano-montmorillonite to form a plant extract composite carrier. A2. Preparation of compound probiotic microcapsules: Compound probiotic powder is encapsulated in microcapsules using trehalose and sodium caseinate as double-layer wall materials, wherein the inner wall material is trehalose and the outer wall material is sodium caseinate, thus obtaining double-layer wall material microcapsules. A3. Low-temperature blending: Under nitrogen protection at 4–10°C, the composite carrier of A1, the microcapsules of A2, xylooligosaccharides, and β-glucan are placed in a three-dimensional mixer for blending for 20–40 minutes to obtain a synergistic additive.

[0023] Specifically, the nanoemulsification in A1 is carried out using a high-pressure homogenization method, which includes: A11. Mix the extracts of *Polygonum multiflorum*, *Eucommia ulmoides*, *Rosemaryia pubescens*, and *Cinnamomum cassia* in proportion, add an ethanol-water mixture to dissolve them completely, and then add 2-5% of Tween-80 as an emulsifier as the total weight of the plant extracts. Stir magnetically at 35-45°C for 30-45 minutes to form a preliminary emulsion, wherein the volume ratio of ethanol to water is 1:3. A12. Transfer the prepared emulsion to a high-pressure homogenizer, set the homogenization pressure to 80-120MPa, the homogenization temperature to 25-35℃, and continuously cycle the homogenization 3-5 times. Detect the particle size of the emulsion using a laser particle size analyzer until more than 90% of the particles are distributed in the range of 50-100nm, and obtain a nano-emulsified plant extract mixture. A13. The prepared nano-emulsified plant extract mixture is stirred at 45°C for 1.5 hours, then freeze-dried under vacuum and pulverized to a particle size of less than 80nm to form a stable plant extract composite carrier.

[0024] The above methods and steps can effectively preserve the live bacteria activity of the compound probiotics and the effective components of the plant extracts, ensuring that the synergistic effect between the components is fully exerted.

[0025] Specifically, the preparation of the double-walled microcapsules in A2 employs a combined spray condensation-freeze-drying technique, which includes: A21. Mix the compound probiotic powder with trehalose in a certain proportion, add an appropriate amount of sterile water and stir until completely dissolved to form a uniform core material-inner wall material mixture. A22. The mixture is atomized by a high-pressure spray device at a spray pressure of 0.3 to 0.5 MPa. The atomized droplets are sprayed into a liquid paraffin condensation medium at -15 to -10°C to form probiotic microspheres that encapsulate the inner wall material. The microspheres are collected and washed 2 to 3 times with sterile anhydrous ethanol to remove residual liquid paraffin on the surface, thus obtaining the preliminary encapsulated microsphere intermediate.

[0026] The above methods and steps can enhance the probiotics' tolerance to high temperatures, stomach acid, and bile salts, ensuring that they can successfully reach the animal's intestines and exert their activity. They can also work synergistically with plant extracts, co-carriers, and protectants to maximize the effectiveness of antibiotic-free aquaculture applications of the additives.

[0027] Specifically, the low-temperature blending of A3 includes: A31. First, premix xylooligosaccharides and β-glucan for 5-10 minutes; A32. Add the plant extract complex carrier and mix for 10-15 minutes. A33. Add the double-layer wall material microcapsules and mix for 5-10 minutes.

[0028] The above steps effectively prevent the microcapsules of the compound probiotics from rupturing due to excessive mechanical shear force during mixing, ensuring the integrity of the microcapsules and the survival rate of live probiotics. Simultaneously, they promote the uniform dispersion of the compound carrier of xylooligosaccharides, β-glucan, and plant extracts, forming a stable synergistic system among the components. This further enhances the protective and synergistic effects of the carrier on the probiotics, ultimately ensuring the synergistic effect of the additive's functional components during application, providing reliable support for improving the intestinal health and growth performance of animals in antibiotic-free farming.

[0029] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.

[0030] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A synergistic additive of antibiotic-free probiotics and plant extracts for aquaculture, characterized in that, The ingredients, by weight, include the following: Compound probiotic powder: 30-50 parts, wherein the compound probiotics include Bifidobacterium animalis subsp. lactis V9, Pediococcus lactis PA-19, Bacillus subtilis BS-2, Clostridium butyricum CB-1 and Saccharomyces cerevisiae SC-22; Synergistic plant extract complex: 20-35 parts, wherein the plant extracts include extracts of *Gnaphalium affine*, *Eucommia ulmoides*, rosemary, and cinnamon; Synergistic carrier: 15-25 parts, including xylooligosaccharides, β-glucan and nano-montmorillonite; Stabilizing and protective agent: 5-10 parts, including trehalose and sodium caseinate.

2. The antibiotic-free compound probiotic and plant extract synergistic additive for aquaculture according to claim 1, characterized in that, The weight parts of each component in the compound probiotic powder are as follows: Bifidobacterium animalis subsp. lactis V9: 10–15 portions; Pediococcus lactis PA-19: 8–12 portions; Bacillus subtilis BS-2: 5-10 parts; Clostridium butyricum CB-1: 5-8 parts; Saccharomyces cerevisiae SC-22: 2 to 5 parts.

3. The antibiotic-free compound probiotic and plant extract synergistic additive for aquaculture according to claim 1, characterized in that, The weight parts of each component in the synergistic plant extract complex are as follows: Extract of *Cephalotaxus fortunei*: 8–12 parts; Eucommia ulmoides extract: 5-8 parts; Rosemary extract: 4-7 parts; Cinnamon extract: 3-5 parts.

4. The antibiotic-free compound probiotic and plant extract synergistic additive for aquaculture according to claim 1, characterized in that, The weight parts of each component in the synergistic carrier are as follows: Xylooligosaccharides: 8–12 parts; β-glucan: 4–7 parts; Nano-montmorillonite: 3-6 parts.

5. The antibiotic-free compound probiotic and plant extract synergistic additive for aquaculture according to claim 1, characterized in that, The weight parts of each component in the stabilizer are as follows: Trehalose: 3-5 parts; Sodium caseinate: 2-5 parts.

6. A method for preparing the additive according to any one of claims 1-5, characterized in that, Includes the following steps: A1. Preparation of synergistic plant extract composite carrier: extracts of Angelica sinensis, Eucommia ulmoides, rosemary, and cinnamon are nanoemulsified and intercalated with nano-montmorillonite to form a plant extract composite carrier. A2. Preparation of compound probiotic microcapsules: Compound probiotic powder is encapsulated in microcapsules using trehalose and sodium caseinate as double-layer wall materials, wherein the inner wall material is trehalose and the outer wall material is sodium caseinate, thus obtaining double-layer wall material microcapsules. A3. Low-temperature blending: Under nitrogen protection at 4–10°C, the composite carrier of A1, the microcapsules of A2, xylooligosaccharides, and β-glucan are placed in a three-dimensional mixer for blending for 20–40 minutes to obtain a synergistic additive.

7. The preparation method of the antibiotic-free compound probiotic and plant extract synergistic additive for aquaculture according to claim 6, characterized in that, The nanoemulsification described in A1 specifically employs a high-pressure homogenization method, including: A11. Mix the extracts of *Polygonum multiflorum*, *Eucommia ulmoides*, *Rosemaryia pubescens*, and *Cinnamomum cassia* in proportion, add an ethanol-water mixture to dissolve them completely, and then add 2-5% of Tween-80 as an emulsifier as the total weight of the plant extracts. Stir magnetically at 35-45°C for 30-45 minutes to form a preliminary emulsion, wherein the volume ratio of ethanol to water is 1:

3. A12. Transfer the prepared emulsion to a high-pressure homogenizer, set the homogenization pressure to 80-120MPa, the homogenization temperature to 25-35℃, and continuously cycle the homogenization 3-5 times. Detect the particle size of the emulsion using a laser particle size analyzer until more than 90% of the particles are distributed in the range of 50-100nm, and obtain a nano-emulsified plant extract mixture. A13. The prepared nano-emulsified plant extract mixture is stirred at 45°C for 1.5 hours, then freeze-dried under vacuum and pulverized to a particle size of less than 80nm to form a stable plant extract composite carrier.

8. The preparation method of the antibiotic-free compound probiotic and plant extract synergistic additive for aquaculture according to claim 6, characterized in that, The preparation of the double-walled microcapsules described in A2 employs a combined spray condensation-freeze-drying technique, specifically including: A21. Mix the compound probiotic powder with trehalose in a certain proportion, add an appropriate amount of sterile water and stir until completely dissolved to form a uniform core material-inner wall material mixture. A22. The mixture is atomized by a high-pressure spray device at a spray pressure of 0.3 to 0.5 MPa. The atomized droplets are sprayed into a liquid paraffin condensation medium at -15 to -10°C to form probiotic microspheres that encapsulate the inner wall material. The microspheres are collected and washed 2 to 3 times with sterile anhydrous ethanol to remove residual liquid paraffin on the surface, thus obtaining the preliminary encapsulated microsphere intermediate.

9. The preparation method of the antibiotic-free compound probiotic and plant extract synergistic additive for aquaculture according to claim 6, characterized in that, The low-temperature blending described in A3 specifically includes: A31. First, premix xylooligosaccharides and β-glucan for 5-10 minutes; A32. Add the plant extract complex carrier and mix for 10-15 minutes. A33. Add the double-layer wall material microcapsules and mix for 5-10 minutes.

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