Feed resistance replacing composition and application thereof
By encapsulating tea polyphenols and gallic acid in quaternized chitosan nanoparticles, and combining them with glucose oxidase and galactooligosaccharides, a multi-level linkage system was constructed, which solved the problem of poor efficacy of existing antibiotic alternatives and achieved safe and efficient growth promotion and disease control effects.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-01-16
- Publication Date
- 2026-04-10
AI Technical Summary
Existing antibiotic alternatives have limited effectiveness, poor stability, and high costs in animal husbandry, making it difficult to completely replace the growth-promoting and disease-controlling effects of antibiotics.
By encapsulating tea polyphenols and gallic acid in quaternized chitosan nanoparticles, combined with glucose oxidase and galactooligosaccharides, a three-level linkage system of "precise antibacterial-environmental regulation-ecological restoration" is constructed. The quaternized chitosan nanoparticles deliver tea polyphenols and gallic acid in a targeted manner, glucose oxidase regulates the intestinal environment, and galactooligosaccharides promote the proliferation of probiotics, thereby achieving a synergistic effect.
It significantly improves the efficiency of active substances, promotes animal growth, improves intestinal health, reduces diarrhea rate, and has an effect close to that of antibiotics, while being safe and easy to promote.
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Abstract
Description
TECHNICAL FIELD
[0001] The present application belongs to the technical field of animal feed additives, and particularly relates to a feed antibiotic replacement composition and application thereof. BACKGROUND
[0002] For a long time, antibiotics have been widely used in the breeding industry to promote animal growth and prevent diseases, but the resulting bacterial drug resistance, drug residues and environmental pollution have become increasingly serious public health problems. The implementation of the "ban" and "limit" policies worldwide has made it an urgent task for the livestock feed industry to find safe and efficient antibiotic substitutes.
[0003] In this context, a variety of types of antibiotic replacement products have emerged in the market, such as organic acids, plant extracts, enzyme preparations, probiotics, etc. However, many products have the disadvantages of single effect, poor stability, unclear mechanism of action or high cost, etc., and it is often difficult to completely replace the growth-promoting and disease-preventing effects of antibiotics in actual application. Therefore, it is of great industrial necessity and application value to develop a new type of composite antibiotic replacement scheme that is safe and healthy, has significant effect, controllable cost and easy to popularize. SUMMARY
[0004] In view of the deficiencies in the prior art, the present application provides a feed antibiotic replacement composition and application thereof, which uses quaternary ammonium salt chitosan nanoparticles to embed tea polyphenols and gallic acid, which can effectively protect the two active substances from damage by gastric acid and digestive enzymes, greatly improving the action efficiency of both.
[0005] To achieve the above-mentioned purposes, the following technical solutions are adopted in the present application: In a first aspect, the present application provides a feed antibiotic replacement composition, comprising the following raw materials in parts by weight per 100 parts: 10-30 parts of quaternary ammonium salt chitosan nanoparticles loaded with tea polyphenols and gallic acid, 5-20 parts of glucose oxidase preparation, 5-15 parts of oligogalactose, and the balance being carrier silicon dioxide.
[0006] In the above technical solution, quaternary ammonium salt chitosan, as a cationic polymer, has antibacterial activity and mucosal adhesion properties; tea polyphenols are a pure natural substance extracted from tea leaves, with broad-spectrum antibacterial activity; gallic acid, as a polyphenolic organic compound, has strong antioxidant properties, and is widely distributed in plants such as gallnut, tea leaves, and oak bark, and has properties such as antioxidant, antibacterial, and anti-inflammatory.
[0007] Glucose oxidase can quickly reduce the local pH value and oxygen partial pressure of the intestinal tract by catalyzing the consumption of oxygen, producing gluconic acid and hydrogen peroxide, and the biochemical action of “oxygen consumption and acid production” can effectively inhibit the proliferation of aerobic pathogenic bacteria (such as Escherichia coli and Salmonella), and create a favorable environment for intestinal microecological regulation. Galacto-oligosaccharides, as a kind of prebiotics, can specifically promote the colonization and proliferation of beneficial bacteria such as Bifidobacterium and Lactobacillus, and the short-chain fatty acids produced by the metabolism can further consolidate the intestinal barrier function and realize the long-term balance and repair of the microecology.
[0008] The core advantage of the present application lies in the construction of a “precision antibacterial-environmental regulation-ecological restoration” three-level linkage mechanism. First, the tea polyphenols and gallic acid released by the quaternary ammonium salt chitosan nanoparticles with antibacterial effect are targeted and delivered, which can directly inhibit pathogenic bacteria; at the same time, the glucose oxidase quickly reduces the local pH value and oxygen partial pressure of the intestinal tract through “oxygen consumption and acid production”, not only inhibits aerobic pathogenic bacteria, but also creates an optimal weak acid environment for the stronger antibacterial activity of polyphenols, producing a functional synergistic amplification effect. On this basis, galacto-oligosaccharides promote the proliferation of probiotics in the optimized microenvironment, and consolidate the replacement resistance effect.
[0009] Preferably, the enzyme activity of the glucose oxidase preparation is 10,000-20,000 U / g.
[0010] Preferably, the mass ratio of tea polyphenols to gallic acid is 1: (2-2.5).
[0011] Preferably, the mass ratio of the sum of tea polyphenols and gallic acid to quaternary ammonium salt chitosan nanoparticles is 1: (1-2.5).
[0012] In the second aspect, the present application provides a preparation method of the feed replacement composition, comprising the following steps: S1, mixing quaternary ammonium salt chitosan and acetic acid aqueous solution to obtain a carrier solution; S2, mixing tea polyphenols, gallic acid and water to obtain an active substance solution; S3, mixing the carrier solution, the active substance solution and the crosslinking agent, then performing solid-liquid separation to obtain a precipitate, and then washing, freeze-drying the precipitate to obtain quaternary ammonium salt chitosan nanoparticles loaded with tea polyphenols and gallic acid; S4, mixing the quaternary ammonium salt chitosan nanoparticles loaded with tea polyphenols and gallic acid, the glucose oxidase preparation, the galacto-oligosaccharides and the carrier silicon dioxide to obtain the feed replacement composition.
[0013] In some embodiments, the specific preparation method is as follows: Quaternized chitosan is dissolved in an aqueous acetic acid solution with a concentration of 0.5-2.0% (v / v) to prepare a carrier solution with a mass-volume concentration of 5-15 mg / mL; tea polyphenols and gallic acid are dissolved together in deionized water at a mass ratio of 1:(2-2.5) to prepare an active ingredient solution with a total mass-volume concentration of 6-10 mg / mL; under magnetic stirring at a speed of 300-600 rpm, the active ingredient solution is slowly added to the carrier solution and mixed; then... A sodium tripolyphosphate aqueous solution with a concentration of 1.0-2.0 mg / mL is slowly added dropwise to the carrier solution as a crosslinking agent. The volume of the crosslinking agent added is 1-1.5 times the volume of the carrier solution. After the addition is complete, the reaction mixture is stirred for 0.5-1 h. After stirring, the reaction mixture is placed in a centrifuge and centrifuged at 10000-15000 rpm for 10-20 min to separate the solid and liquid phases and collect the precipitate. The precipitate is washed 2 to 3 times with deionized water. The washed precipitate is then freeze-dried to obtain polyphenol-loaded nanoparticle powder. The obtained nanoparticle powder is mixed evenly with glucose oxidase preparation, galactooligosaccharides, and the carrier according to the proportions described in the technical solution to obtain the feed antibiotic alternative composition.
[0014] Preferably, in step S1, the concentration of the acetic acid aqueous solution is 0.5-2.0% (v / v); and / or, In step S1, the mass-volume concentration of the carrier solution is 5-15 mg / mL.
[0015] Preferably, in step S2, the mass-volume concentration of the active ingredient solution is 6-10 mg / mL.
[0016] Preferably, in step S3, the crosslinking agent is an aqueous solution of sodium tripolyphosphate with a concentration of 1.0-2.0 mg / mL; and / or, In step S3, the volume of the crosslinking agent added is 1-1.5 times the volume of the carrier solution.
[0017] Thirdly, the present invention provides the application of the aforementioned feed antibiotic alternative composition, or the feed antibiotic alternative composition prepared by the aforementioned preparation method, in the preparation of animal feed.
[0018] Preferably, the amount of the feed antibiotic alternative composition added to the animal feed is 0.01-0.05 wt%.
[0019] Compared with the prior art, the beneficial effects of the present invention are as follows: (1) This invention scientifically combines tea polyphenols and gallic acid, which are protected by being encapsulated in quaternized chitosan nanocarriers, with glucose oxidase, which can effectively change the intestinal microenvironment, and galactooligosaccharides, which can promote the proliferation of beneficial bacteria, to construct a linkage system of "precise antibacterial, environmental regulation and ecological restoration". This system can produce a synergistic amplification effect in antibacterial and intestinal health maintenance, and effectively replace antibiotics.
[0020] (2) The present invention uses quaternized chitosan nano-encapsulated tea polyphenols and gallic acid, which can effectively protect the two active substances from the destruction of gastric acid and digestive enzymes, and greatly improve their efficiency.
[0021] (3) The preparation process of the antibiotic alternative composition in this invention avoids the technical difficulties of complex co-encapsulation of polyphenols and enzymes. The core steps are simplified to the preparation of nanoparticles and the physical mixing of all components. This process route has low equipment requirements, is easy to operate, and has a short production cycle, which is conducive to large-scale production and its application in the feed industry.
[0022] (4) The functional ingredients selected in this invention have the advantages of being safe, compliant, healthy, and free of toxic side effects, providing a reliable guarantee for the production of green and safe animal products. Detailed Implementation
[0023] The present invention will now be described in further detail with reference to specific embodiments, so that those skilled in the art can more clearly understand the present invention.
[0024] Example 1 This embodiment provides a feed antibiotic alternative composition, which is uniformly mixed from the following components in parts by weight: 15 parts of quaternized chitosan nanoparticles loaded with tea polyphenols and gallic acid, 8 parts of glucose oxidase preparation (enzyme activity: 20000U / g), 7 parts of galactooligosaccharide, and 70 parts of feed-grade silica carrier.
[0025] The specific preparation method of the feed antibiotic alternative composition includes the following steps: (1) Preparation of nanocarrier particles: 1.0 g of quaternized chitosan was dissolved in 100 mL of 1.0% (v / v) acetic acid solution to obtain a carrier solution. 0.3 g of tea polyphenols and 0.6 g of gallic acid were dissolved together in 100 mL of deionized water to obtain an active solution. The active solution was added dropwise to the carrier solution under magnetic stirring (500 rpm), and after mixing, 100 mL of 1.0 mg / mL sodium tripolyphosphate crosslinking agent aqueous solution was slowly added, and the reaction was continued for 1 hour. The reaction solution was separated by centrifugation (12000 rpm, 15 min), and the precipitate was washed twice with deionized water and then freeze-dried to obtain nanoparticle powder.
[0026] (2) Composition mixing: Weigh each component according to the proportions in Example 1 above, mix them evenly, and the composition is obtained.
[0027] Example 2 This embodiment provides a feed antibiotic alternative composition, which is uniformly mixed from the following components in parts by weight: 20 parts of quaternized chitosan nanoparticles loaded with tea polyphenols and gallic acid, 12 parts of glucose oxidase preparation (enzyme activity: 20000U / g), 10 parts of galactooligosaccharide, and 58 parts of feed-grade silica carrier.
[0028] The specific preparation method of the feed antibiotic alternative composition is the same as that in Example 1.
[0029] Example 3 This embodiment provides a feed antibiotic alternative composition, which is uniformly mixed from the following components in parts by weight: 25 parts of quaternized chitosan nanoparticles loaded with tea polyphenols and gallic acid, 15 parts of glucose oxidase preparation (enzyme activity: 20000U / g), 8 parts of galactooligosaccharides, and 52 parts of feed-grade silica carrier.
[0030] The specific preparation method of the feed antibiotic alternative composition is the same as that in Example 1.
[0031] Comparative Example 1 This comparative example provides a feed antibiotic alternative composition, with the same formulation and preparation method as Example 1, except that the quaternary ammonium salted chitosan nanoparticles loaded with tea polyphenols and gallic acid are replaced with tea polyphenols.
[0032] Comparative Example 2 This comparative example provides a feed antibiotic alternative composition, with the same formulation and preparation method as Example 1, except that the quaternary ammonium salted chitosan nanoparticles loaded with tea polyphenols and gallic acid are replaced with gallic acid.
[0033] Comparative Example 3 This comparative example provides a feed antibiotic alternative composition. The formulation and preparation method are the same as those in Example 1. The difference is that the quaternary ammonium salted chitosan nanoparticles loaded with tea polyphenols and gallic acid are replaced with a mixture of tea polyphenols and gallic acid (mass ratio is the same as in Example 1).
[0034] Comparative Example 4 This comparative example provides a feed antibiotic alternative composition, with the same formulation and preparation method as Example 1, except that the quaternary ammonium salted chitosan nanoparticles loaded with tea polyphenols and gallic acid are replaced with oxytetracycline hydrochloride.
[0035] Performance Tests and Results The Oxford cup method and the double broth dilution method were used to determine the antibacterial activity (inhibition zone diameter) and minimum inhibitory concentration (MIC) of the feed antibiotic alternatives prepared in Examples 1-3 and Comparative Examples 1-4 against Escherichia coli and Staphylococcus aureus, respectively. The results are shown in Table 1.
[0036] Table 1. Evaluation results of antibacterial effects of Comparative Examples 1-4 and Examples 1-3
[0037] As shown in Table 1, the antibacterial properties of the feed antibiotic alternative compositions prepared in Examples 1-3 were significantly better than those in Comparative Examples 1-3, and the effects were close to those in Comparative Example 4 (antibiotic group). Among them, the antibiotic alternative composition in Example 3 had the strongest antibacterial activity, with a MIC value significantly lower than that in Comparative Examples 1-3 and no significant difference from the antibiotic group, indicating that the combination of polyphenol-chitosan nanoparticles with glucose oxidase and galactooligosaccharides produced a significant synergistic antibacterial effect.
[0038] Application Example 1 (Health Benefits of Routine Feeding) Five hundred and sixty healthy AA broilers of similar weight and good health at one day old were selected and randomly divided into seven groups, with five replicates per group and sixteen broilers (half male and half female) per replicate. The treatment design is shown in Table 2. The experimental period was 42 days. During the entire experimental period, each treatment group was fed the corresponding diet and the broilers were immunized and fed according to the conventional feeding program.
[0039] Table 2 Experimental Design
[0040] After the experiment, growth performance and cecal flora indicators were collected, and the results are shown in Table 3.
[0041] Table 3. Growth performance and gut microbiota results of broilers under conventional feeding (1-42 days old)
[0042] As shown in Table 3, compared with the blank control group, the growth performance (daily weight gain, feed conversion ratio) and intestinal flora (increased lactic acid bacteria, decreased Escherichia coli) of each treatment group were improved. The effects of Examples 1-3 were more significant, all of which were significantly higher than the blank control group, the enzyme preparation group, and the polyphenol mixture group. Moreover, the growth performance of Example 3 was comparable to that of the antibiotic control group, and it was even better than the antibiotic group in inhibiting harmful Escherichia coli and promoting beneficial lactic acid bacteria.
[0043] The above results demonstrate that the optimized formulation represented in Example 3, under conventional feeding conditions, not only replaces antibiotics in promoting growth and improving feed efficiency, but also exhibits a better "health-preserving" effect than antibiotics in optimizing the gut microbiota (significantly increasing beneficial bacteria and inhibiting harmful bacteria), a key health indicator. Furthermore, the effects of all example groups are significantly better than those of single-component or simple mixture groups, fully verifying the high efficiency of the "multi-component synergistic design" in this invention, providing an efficient and safe solution for antibiotic-free aquaculture.
[0044] Application Example 2 (Virus Attack Protection Effect) Avian colibacillosis is one of the most common and frequently occurring diseases in poultry farming. This series of diseases caused by *Escherichia coli* encompasses numerous symptoms, including granulomatosis, peritonitis, salpingitis, omphalitis, synovitis, air sacculitis, conjunctivitis, and yolk sac peritonitis, posing a significant threat to the poultry industry. Therefore, this experiment used *E. coli* challenge to evaluate the protective effect of the composition of this invention against viral infection.
[0045] Sixty-fourty healthy AA broiler chickens aged one day were randomly divided into eight treatment groups (one additional challenge control group plus a basal diet was added; the design of other groups was the same as in Application Example 1), with five replicates per group and 16 chickens per replicate. All groups were fed their corresponding experimental diets from day 1 to day 14. On day 15, all chickens except the blank control group were subjected to E. coli bacterial suspension (1×10⁻⁶). 9 CFU / feather was administered via intraperitoneal injection to challenge the virus. After challenge, the birds were fed the same diet until 42 days of age, at which point the diarrhea rate was recorded 15-21 days post-challenge. Growth performance indicators were calculated after the experiment, and cecal microbiota were measured. The results are shown in Table 4 below.
[0046] Table 4. Growth performance and cecal flora of broilers under challenge conditions (1-42 days old)
[0047] Table 4 shows that, compared with the challenge control group, the antibiotic group, enzyme group, polyphenol combination group, and groups 1-3 all improved the growth performance and intestinal flora of broilers after challenge. Among them, group 3 showed the best effect, with growth performance improvement comparable to the antibiotic group and intestinal flora improvement even better than the antibiotic group, and significantly better than the enzyme group and polyphenol combination group. Regarding diarrhea rate, the diarrhea rate of broilers increased significantly after challenge, while groups 1-3 significantly reduced the diarrhea rate, achieving an effect comparable to that of antibiotics. These results indicate that, under challenge conditions, the composition of the present invention can effectively alleviate growth inhibition, decreased feed efficiency, and severe diarrhea in broilers caused by challenge, and has a prominent role in improving intestinal flora and promoting the growth of beneficial bacteria.
[0048] This invention, from the perspective of "active antibacterial activity of composite polyphenol nanoparticles, synergistic effect of enzyme preparations altering the intestinal microenvironment, and prebiotics promoting the restoration of intestinal microecology," scientifically combines tea polyphenols and gallic acid encapsulated and protected by quaternized chitosan nanocarriers with glucose oxidase, which can specifically alter the intestinal microenvironment, and galactooligosaccharides, which promote the proliferation of beneficial bacteria. Its beneficial effects are comparable to those of antibiotics and are significantly superior to single enzyme preparations or physical mixtures of polyphenols. This indicates that the antibiotic alternative composition of this invention has a good synergistic effect and outstanding efficacy, and can provide strong data support and direction for the problem of antibiotic alternatives in the breeding and feed industries.
[0049] The above examples are merely illustrative of the present invention and do not constitute a limitation on the scope of protection of the present invention. All designs that are the same as or similar to the present invention are within the scope of protection of the present invention.
Claims
1. A feed antibiotic alternative composition, characterized in that, Based on 100 parts, the raw materials include the following parts by weight: 10-30 parts of quaternary ammonium salted chitosan nanoparticles loaded with tea polyphenols and gallic acid, 5-20 parts of glucose oxidase preparation, 5-15 parts of galactooligosaccharides, and the balance being carrier silica.
2. The feed antibiotic alternative composition according to claim 1, characterized in that, The enzyme activity of glucose oxidase preparations is 10,000-20,000 U / g.
3. The feed antibiotic alternative composition according to claim 1, characterized in that, The mass ratio of tea polyphenols to gallic acid is 1:(2-2.5).
4. The feed antibiotic alternative composition according to claim 1, characterized in that, The mass ratio of the sum of tea polyphenols and gallic acid to the mass of quaternized chitosan nanoparticles is 1:(1-2.5).
5. The method for preparing the feed antibiotic alternative composition according to any one of claims 1-4, characterized in that, Includes the following steps: S1. Mix quaternized chitosan and an aqueous acetic acid solution to obtain a carrier solution; S2. Mix tea polyphenols, gallic acid and water to obtain an active ingredient solution; S3. After mixing the carrier solution, active substance solution and crosslinking agent, the precipitate is separated by solid-liquid separation, washed and freeze-dried to obtain quaternary ammonium salted chitosan nanoparticles loaded with tea polyphenols and gallic acid. S4. Mix quaternized chitosan nanoparticles loaded with tea polyphenols and gallic acid, glucose oxidase preparation, galactooligosaccharides and carrier silica to obtain a feed antibiotic alternative composition.
6. The preparation method according to claim 5, characterized in that, In step S1, the concentration of the acetic acid aqueous solution is 0.5-2.0% (v / v); and / or, In step S1, the mass-volume concentration of the carrier solution is 5-15 mg / mL.
7. The preparation method according to claim 5, characterized in that, In step S2, the mass-volume concentration of the active ingredient solution is 6-10 mg / mL.
8. The preparation method according to claim 5, characterized in that, In step S3, the crosslinking agent is an aqueous solution of sodium tripolyphosphate with a concentration of 1.0-2.0 mg / mL; and / or, In step S3, the volume of the crosslinking agent added is 1-1.5 times the volume of the carrier solution.
9. The use of the feed antibiotic alternative composition according to claim 1, or the feed antibiotic alternative composition prepared by the preparation method according to any one of claims 2-8, in the preparation of animal feed.
10. The application according to claim 9, characterized in that, The amount of the feed antibiotic alternative composition added to animal feed is 0.01-0.05 wt%.