A premix for preventing and treating chicken gastric adenitis, a preparation method and application thereof

Abstract

The application provides a premix for preventing and treating chicken gastritis, a preparation method and application thereof, and relates to the technical field of animal feed. Specifically, the premix comprises a composite probiotic microcapsule, a composite enzyme preparation, a functional excipient and a carrier; the functional excipient comprises a nano-modified silico-aluminate, a jerusalem artichoke powder, a curcumin microcapsule and vitamin E succinate; the composite probiotic microcapsule is prepared by drying a W / O / W emulsion, and the inner water phase of the W / O / W emulsion comprises a probiotic composite agent. The application realizes the prevention and treatment of chicken gastritis through a multi-target intervention strategy system based on the synergistic effect of bacteria and enzymes, and provides an efficient, safe and economical gastritis prevention and treatment premix through a three-fold synergistic mechanism of "toxin biodegradation-pathogenic bacteria inhibition-mucosal barrier repair", a three-section microcapsule embedding technology and a low-temperature solid-state activation process in the preparation method, and material modification and preparation engineering technology, and solves the compatibility problem of components innovatively.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of animal feed, in particular to a premix for preventing and treating chicken gastritis and a preparation method and application thereof. BACKGROUND

[0002] Chicken gastritis is a common digestive tract disease in intensive chicken farming, which is characterized by hyperemia, edema, erosion and functional disorder of the gastric mucosa. In the middle and late stages of the disease, the sick chickens are extremely emaciated and die due to exhaustion. With the increase of breeding density and the fluctuation of feed raw materials, the incidence of the disease has shown a significant upward trend. The pathogenic factors of gastritis are complex, including mycotoxin pollution, pathogenic microorganism infection and feeding management factors, etc. In recent years, the synergistic pathogenic effect of mycotoxin and pathogenic microorganism has been widely concerned, which forms a vicious cycle of "toxin-microorganism-inflammation".

[0003] At present, the prevention and treatment methods for broiler chicken gastritis mainly include chemical drug prevention and treatment (antibiotic treatment, antifungal drugs, proton pump inhibitors, etc.), biological agent application (single probiotic preparation, enzyme preparation, toxin adsorbent, etc.), nutritional regulation means (acidifying agent, mucosa repair agent, immune enhancer, etc.). However, the existing technology at least has the following defects: (1) single mechanism: unable to simultaneously solve the three core problems of toxin removal, microbial balance and mucosa repair; (2) loss of biological activity: gastric acid environment and feed processing cause the inactivation of probiotics and enzyme preparations; (3) lack of synergistic effect: physical or chemical antagonism exists between toxin adsorbent and biological agent, such as montmorillonite adsorbing probiotics to reduce their colonization efficiency. In view of the above defects, it is urgent to propose a new method for preventing and treating broiler chicken gastritis, which provides a reference for clinical and production practice; and feed is an indispensable component in broiler farming, and if the feed or its derivatives (such as premix) can be reasonably used to prevent and treat gastritis, the production performance and economic benefits of chickens will be effectively improved.

[0004] Therefore, the present application is proposed. SUMMARY

[0005] The first object of the present application is to provide a premix for preventing and treating chicken gastritis, which is mainly used to solve the technical defects that the existing prevention and treatment methods for chicken gastritis cannot simultaneously solve the single mechanism, loss of biological activity of the preparation and lack of synergistic effect.

[0006] The second object of the present application is to provide a preparation method of the premix for preventing and treating chicken gastritis.

[0007] The third object of the present application is to provide an animal feed for preventing and treating gastritis.

[0008] Research has shown that the bacterial-enzyme synergistic system has promising technical prospects for preventing proventriculitis in chickens. Specifically, the bacterial-enzyme synergistic system of this invention comprises three parts: targeted degradation, microecological remodeling, and mucosal repair. First, targeted degradation uses specific enzymes to convert mycotoxin contamination, one of the pathogenic factors, into low-toxicity metabolites (e.g., aflatoxin B1 is converted by aflatoxin-degrading enzyme in this invention). Targeted degradation also further neutralizes the toxin activity through reduced glutathione produced by probiotic metabolism. Second, microecological remodeling uses specific probiotic combinations to inhibit or eliminate pathogens (e.g., extracellular polysaccharides secreted by *Saccharomyces boulardii* can inhibit *Helicobacter pylori* biofilm formation, and its combination with *Bacillus subtilis* can increase the pathogen clearance rate by 3 times). Third, mucosal repair uses microorganisms to regenerate or repair the proventricular mucosa of chicken lesions (e.g., γ-aminobutyric acid produced by *Pediococcus lactis* promotes mucus layer regeneration in this invention, and its synergy with curcumin can increase the ulcer healing speed by 50%). However, the efficient synergy of bacteria and enzymes in the aforementioned bacterial-enzyme synergistic system still faces other key technical challenges, such as: the optimal pH of enzymes is usually neutral, which does not match the pH of 2.0 to 3.5 in the acidic environment of chicken stomach; high temperatures (75℃ to 90℃) and high shear forces in feed processing can easily lead to the inactivation of bioactive components; and there are compatibility issues between toxin adsorbents and bacterial enzymes, as traditional adsorbents may non-specifically adsorb beneficial components.

[0009] In order to achieve the above-mentioned objectives of the present invention, the following technical solution is adopted:

[0010] A premix for preventing proventriculitis in chickens includes: compound probiotic microcapsules, compound enzyme preparation, functional excipients, and carrier;

[0011] The functional excipients include nano-modified aluminosilicate, Jerusalem artichoke powder, curcumin microcapsules, and vitamin E succinate.

[0012] The compound probiotic microcapsules are prepared by drying a W / O / W emulsion, wherein the W / O / W emulsion comprises an outer aqueous phase, an intermediate oil phase, and an inner aqueous phase, and the inner aqueous phase comprises a probiotic compound.

[0013] A method for preparing a premix for preventing proventriculitis in chickens includes the following steps:

[0014] The components of the intermediate oil phase and the inner aqueous phase are mixed and homogenized at 8000 rpm to 15000 rpm until homogeneous. Then, the outer aqueous phase is added and homogenized at 4000 rpm to 8000 rpm until homogeneous to obtain a W / O / W emulsion. The W / O / W emulsion is then subjected to low-temperature spray drying to obtain composite probiotic microcapsules.

[0015] The premix precursor is obtained by mixing compound probiotic microcapsules, compound enzyme preparation, functional excipients and carrier through four-stage amplification. The premix precursor is then sealed and activated at 30℃~40℃ to obtain a premix for the prevention and treatment of proventriculitis in chickens.

[0016] An animal feed for preventing proventriculitis, comprising the aforementioned premix for preventing proventriculitis in chickens.

[0017] Compared with existing technologies, the beneficial effects of this invention are as follows: This invention provides a premix for the prevention and treatment of proventriculitis in chickens based on the binding effect of bacteria and enzymes, especially showing excellent preventive and therapeutic effects for proventriculitis in broilers; this invention achieves a three-level technical effect of "toxin degradation - microbial regulation - mucosal repair" through the spatiotemporal synergy of aflatoxin-degrading enzymes and triple probiotics; and it uses nano-modified aluminosilicate as a directional adsorption carrier, whose surface-grafted amino functional groups can specifically bind to the carbonyl structure of aflatoxin B1; through a three-stage microencapsulation process, it ensures that the survival rate of bacteria and enzymes in the gastric acid environment is >85% and that they are released in the intestine in a targeted manner. This invention breaks through the single-action mode of traditional prevention and treatment methods, and provides a brand-new solution for the prevention and control of proventriculitis in chickens through the interdisciplinary integration of materials science, microbial engineering, and enzyme engineering, with significant economic and social benefits. Detailed Implementation

[0018] The technical solution of the present invention will be clearly and completely described below with reference to specific embodiments. However, those skilled in the art will understand that the embodiments described below are some embodiments of the present invention, but not all embodiments, and are only used to illustrate the present invention, and should not be regarded as limiting the scope of the present invention. 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. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall be followed. Where the manufacturers of reagents or instruments are not specified, they are all conventional products that can be purchased commercially. In addition, the terms "first," "second," and "third" are used for descriptive purposes only and should not be construed as indicating or implying relative importance.

[0019] The first aspect of the present invention is to provide a premix for preventing proventriculitis in chickens.

[0020] The premix comprises the following components: compound probiotic microcapsules, compound enzyme preparation, functional excipients, and carrier; wherein, the functional excipients include nano-modified aluminosilicate, Jerusalem artichoke powder, curcumin microcapsules, and vitamin E succinate; the compound probiotic microcapsules are prepared by drying a W / O / W emulsion, wherein the W / O / W emulsion comprises an outer aqueous phase, an intermediate oil phase, and an inner aqueous phase, wherein the inner aqueous phase comprises a probiotic compound.

[0021] In a preferred embodiment, the premix comprises the following components by weight: 120-180 parts of compound probiotic microcapsules, 60-100 parts of compound enzyme preparation, 250-330 parts of functional excipients, and 390-570 parts of carrier. In some optional embodiments, the mass fractions of the premixed ingredients include, but are not limited to: compound probiotic microcapsules 120, 125, 130, 140, 150, 160, 170, 175, 180; compound enzyme preparations 60, 65, 70, 80, 90, 95, 100; functional excipients 250, 260, 270, 280, 290, 300, 310, 320, 330; and carriers 390, 400, 425, 450, 475, 500, 525, 550, 570. The above mass fractions can be any of the listed point values ​​or any range of values ​​formed by any two point values.

[0022] In a preferred embodiment, the compound probiotic microcapsules, the compound enzyme preparation, and the functional excipients are loaded onto the pores or surface of the carrier.

[0023] In one preferred embodiment, the probiotic complex includes Bacillus subtilis, Saccharomyces boulardii, and Pediococcus lactis; in some more preferred embodiments, the number of live bacteria in the probiotic complex (or in the probiotic microcapsules) includes: Bacillus subtilis ≥ 5.0 * 10⁻⁶. 10 CFU / g, *Saccharomyces boulardii* ≥ 2.0*10 10 CFU / g, Pediococcus lactis ≥1.0*10 11 CFU / g.

[0024] In the compound probiotic microcapsules, the W / O / W emulsion conforms to a well-known definition, namely a water-in-oil-in-water emulsion, satisfying the structural characteristics of an outer continuous aqueous phase, an intermediate dispersed oil phase, and an inner dispersed aqueous phase.

[0025] In a preferred embodiment, the outer aqueous phase includes chitosan, which contributes to the stability of the entire emulsion system. In some more preferred embodiments, the chitosan content in the outer aqueous phase is 0.08 wt.% to 0.3 wt.%, and the solvent of the outer aqueous phase is an aqueous solution of acetic acid with a concentration of 0.8 wt.% to 3 wt.%.

[0026] In a preferred embodiment, the intermediate oil phase includes at least one of monoglyceride, polyglycerol ester, or lecithin, and one of beeswax or shellac; it should be noted that when shellac is used, a plasticizer (such as triethyl citrate) needs to be added as an adjunct. The intermediate oil phase encapsulates the inner aqueous phase, forming an encapsulating protection for the intermediate oil phase and forming oil droplets dispersed in the outer aqueous phase; in some more preferred embodiments, the mass ratio of the monoglyceride to the beeswax is (2-5):1.

[0027] In a preferred embodiment, the inner aqueous phase includes a probiotic complex, and further includes at least one of trehalose or mannitol, and at least one of glutamine or arginine; in some more preferred embodiments, when the following components are used in the inner aqueous phase: the content of trehalose is 8 wt.% to 15 wt.%, the content of mannitol is 28 wt.% to 35 wt.%, the content of glutamine and / or the content of arginine is 3 wt.% to 8 wt.%, and the amount of probiotic complex can be adjusted adaptively according to the bacterial activity provided above.

[0028] In a preferred embodiment, the volume ratio of the outer aqueous phase, the intermediate oil phase, and the inner aqueous phase in the W / O / W emulsion is (8-12):(3-5):1.

[0029] In a preferred embodiment, the compound enzyme preparation includes aflatoxin-degrading enzyme, acidic protease, and β-glucanase, wherein the β-glucanase is a heat-resistant variety that can maintain high catalytic activity under high temperature conditions (≥50℃).

[0030] In a more preferred embodiment, the enzyme activity of the compound enzyme preparation includes: aflatoxin degrading enzyme ≥15000U / g, acidic protease ≥25000U / g, and β-glucanase ≥18000U / g.

[0031] In a preferred embodiment, the functional excipient comprises the following components by weight: 18-25 parts of nano-modified aluminosilicate, 4-7 parts of Jerusalem artichoke powder, 2.5-4 parts of curcumin microcapsules, and 0.8-1.5 parts of vitamin E succinate. In some optional embodiments, the weight parts of each component in the functional excipient include, but are not limited to: 18, 19, 20, 21, 22, 23, 24, 25 parts of nano-modified aluminosilicate; 4, 4.5, 5, 5.5, 6, 6.5, 7 parts of Jerusalem artichoke powder; 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4 parts of curcumin microcapsules; and 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5 parts of vitamin E succinate. The above weight parts can be any of the listed point values ​​or any range of values ​​formed by any two point values.

[0032] It is worth noting that the weight parts of the functional excipients and the premix are measured independently, and the two do not use the same mass part ratio dimension; that is, those skilled in the art should not assume that when the weight of the compound probiotic microcapsules is 120 to 180 parts, the weight of the nano-modified aluminosilicate is 18 to 25 parts.

[0033] In a preferred embodiment, the nano-modified aluminosilicate is obtained by surface amylation modification of aluminosilicate. In this invention, the specific surface area of ​​conventional aluminosilicate can be significantly increased by modifying it, and a sub-nanometer mesoporous structure can be formed, which enhances the specific adsorption of aflatoxin by aluminosilicate. In some more preferred embodiments, the particle size of the nano-modified aluminosilicate is 80 nm to 200 nm.

[0034] In some optional embodiments, the preparation method of the nano-modified aluminosilicate can be as follows: Aluminosilicate or a raw material with aluminosilicate as the main component (such as ZSM-5, Y-type zeolite, etc.) is thoroughly mixed with a silane coupling agent (such as KH-550, APTES, AEAPTMS, etc.) in an organic solvent, and then separated to obtain the nano-modified aluminosilicate. As a more preferred embodiment, the preparation method of the nano-modified aluminosilicate includes: mixing aluminosilicate with an ethanol solution of γ-aminopropyltriethoxysilane, controlling the solid-liquid ratio at 1:8–12 (g / mL), performing ultrasonic treatment, washing, and drying to obtain the nano-modified aluminosilicate.

[0035] In a preferred embodiment, the curcumin microcapsules are a formulation in which curcumin is encapsulated in tiny capsules using microencapsulation technology, which can effectively improve the stability, solubility, bioavailability, and targeting of curcumin; the curcumin content in the curcumin microcapsules is 20 wt.% to 25 wt.%. In some more preferred embodiments, the capsule membrane of the curcumin microcapsules includes hydrogenated palm oil, ethyl cellulose, gum arabic, or gelatin, but hydrogenated palm oil is more preferably used as the capsule shell material.

[0036] In a preferred embodiment, the carrier comprises at least one of defatted rice bran, wheat bran, or corn cob powder; the particle size of the carrier is 60-100 mesh, the porosity of the carrier is 50%-65%, and the specific surface area of ​​the carrier is 3 m². 2 / g~5m 2 / g.

[0037] A second aspect of the present invention is to provide a method for preparing a premix for preventing proventriculitis in chickens as described in the first aspect. The preparation method includes the following steps:

[0038] (1) The components of the intermediate oil phase and the inner aqueous phase are mixed and homogenized at 8000 rpm to 15000 rpm until uniform. Then the outer aqueous phase is added and homogenized at 4000 rpm to 8000 rpm until uniform to obtain a W / O / W emulsion. The W / O / W emulsion is spray-dried at low temperature to obtain composite probiotic microcapsules.

[0039] (2) The compound probiotic microcapsules, compound enzyme preparation, functional excipients and carrier are mixed through four-stage amplification to obtain a premix precursor. The premix precursor is sealed and activated at 30℃~40℃ to obtain a premix for preventing and treating proventriculitis in chickens.

[0040] In a preferred embodiment, the inlet air temperature of the low-temperature spray drying is 40℃~60℃, the outlet air temperature of the low-temperature spray drying is 20℃~30℃, and the atomization pressure of the low-temperature spray drying is 0.7MPa~0.85MPa.

[0041] In a preferred embodiment, the preparation method further includes a pretreatment of the carrier, comprising the following steps: thoroughly mixing the carrier with the oil to reduce the dust of the carrier; in some more preferred embodiments, the oil includes, but is not limited to, soybean oil, corn oil, sunflower seed oil, wheat germ oil, etc.; in some more preferred embodiments, the mass ratio of the oil to the carrier is 0.5% to 3%.

[0042] In a preferred embodiment, the four-stage amplification mixing includes the following steps: S1, thoroughly mixing nano-modified aluminosilicate, Jerusalem artichoke powder and carrier; S2, thoroughly mixing the mixture from S1 with the compound enzyme preparation; S3, thoroughly mixing the mixture from S2, the compound probiotic microcapsules and the curcumin microcapsules; S4, spraying vitamin E succinate onto the mixture from S3.

[0043] In a preferred embodiment, the relative humidity of the activation environment is 80% to 90%.

[0044] In a preferred embodiment, the activation time is 10h to 15h.

[0045] A third aspect of the present invention is to provide an animal feed for preventing proventriculitis, comprising the premix for preventing proventriculitis in chickens as described in the first aspect. It is understood that the animal feed should also include nutrients to meet the specific needs of animal growth, production, and maintenance of vital activities, including but not limited to energy feeds, protein feeds, and roughage. The present invention does not impose any restrictions on the specific composition of the animal feed. In some preferred embodiments, the mass ratio of the premix to the feed is 1.2% to 1.5%.

[0046] Example 1

[0047] (1) Preparation of nano-modified hydrated aluminosilicate:

[0048] Hydrated aluminosilicate was immersed in an ethanol solution of 3 wt.% APTES (γ-aminopropyltriethoxysilane) with a solid-liquid ratio of 1:10 (g / mL); it was ultrasonically treated at 60℃ for 2 h, then centrifuged and washed until neutral, and dried at 120℃ to obtain nano-modified hydrated aluminosilicate.

[0049] (2) Preparation of compound probiotic microcapsules:

[0050] (2-1) Preparation of W / O / W double emulsion: Formulate a probiotic complex (including: Bacillus subtilis 6.0*10) 10 CFU / g, Saccharomyces boulardii 2.5*10 10 CFU / g, Pediococcus lactis 2.0*10 11 Take 200 g / L of probiotic complex, add 10 wt.% trehalose and 5 wt.% glutamine to obtain the inner aqueous phase; prepare monoglycerides and beeswax (mass ratio 3:1), melt them and cool to 45°C to obtain the oil phase; prepare an aqueous solution of 1 wt.% acetic acid, add 0.1 wt.% chitosan to obtain the outer aqueous phase;

[0051] During primary emulsification, the internal aqueous phase and oil phase are mixed at a volume ratio of 1:4 and homogenized at 10,000 rpm for 3 minutes. During secondary emulsification, the primary emulsion and external aqueous phase are mixed at a volume ratio of 1:2 and homogenized at 6,000 rpm for 2 minutes.

[0052] (2-2) Low-temperature spray drying: The homogenized emulsion is placed in a spray drying device, and the inlet air temperature is set to 45℃, the outlet air temperature to 28℃, and the atomization pressure to 0.8MPa.

[0053] (3) Raw material preparation:

[0054] Compound enzyme preparation: aflatoxin degrading enzyme 16000U / g, acidic protease 25000U / g, thermostable β-glucanase 18500U / g;

[0055] Functional excipients: 200g of nano-modified hydrated aluminosilicate, 50g of Jerusalem artichoke powder, 30g of curcumin microcapsules, and 10g of vitamin E succinate;

[0056] 150g of compound probiotic microcapsules, 90g of compound enzyme preparation, 290g of functional excipients, and 470g of defatted rice bran (80 mesh) as carrier.

[0057] (4) Preparation of premix:

[0058] (4-1) Carrier pretreatment: defatted rice bran that has passed through an 80-mesh sieve is premixed with 1 wt.% soybean oil.

[0059] (4-2) Stepwise scaling up and mixing: First, mix nano-modified hydrated aluminosilicate and Jerusalem artichoke powder evenly; then add compound enzyme preparation and mix evenly; then add compound probiotic microcapsules and curcumin microcapsules and mix evenly; finally add vitamin E succinate for spraying.

[0060] (4-3) Solid activation: The mixture obtained in the previous step is treated in a closed environment at 35°C and 85% relative humidity for 12 hours to obtain the premix of this embodiment.

[0061] Example 2

[0062] It is basically the same as Example 1, except that some features are replaced in step (3): 180g of compound probiotic microcapsules, 60g of compound enzyme preparation, 290g of functional excipients, and 470g of carrier defatted rice bran.

[0063] Example 3

[0064] The same as in Example 1, except that some features are replaced in step (3): 120g of compound probiotic microcapsules, 100g of compound enzyme preparation, 290g of functional excipients, and 490g of carrier defatted rice bran.

[0065] Example 4

[0066] It is basically the same as Example 1, except that some features are replaced in step (3): the functional excipients include: 190g of nano-modified hydrated aluminosilicate, 60g of Jerusalem artichoke powder, 28g of curcumin microcapsules, and 12g of vitamin E succinate.

[0067] Example 5

[0068] It is basically the same as Example 1, except that in step (4-3), some features are replaced with: treatment in a closed environment at 40°C and 88% relative humidity for 11 hours.

[0069] Comparative Example 1

[0070] It is basically the same as Example 1, except that step (1) is cancelled and conventional hydrated aluminosilicate is used instead of nano-modified aluminosilicate.

[0071] Comparative Example 2

[0072] The results are basically the same as in Example 1, except that Jerusalem artichoke powder is removed and the mass ratio of the functional excipients is replaced with: 240g of nano-modified hydrated aluminosilicate, 38g of curcumin microcapsules, and 12g of vitamin E succinate.

[0073] Comparative Example 3

[0074] The results are basically the same as in Example 1, except that: the addition of curcumin microcapsules is omitted, and the mass ratio of the functional excipients is replaced with: 220g of nano-modified hydrated aluminosilicate, 58g of Jerusalem artichoke powder, and 12g of vitamin E succinate.

[0075] Comparative Example 4

[0076] The results are basically the same as in Example 1, except that: the addition of vitamin E succinate is omitted, and the mass ratio of the functional excipients is replaced with: 208g of nano-modified hydrated aluminosilicate, 51g of Jerusalem artichoke powder, and 31g of curcumin microcapsules.

[0077] Comparative Example 5

[0078] The process is basically the same as in Example 1, except that step (2) is omitted and the probiotic complex in (2-1) is used to replace the compound probiotic microcapsules.

[0079] Test case

[0080] The therapeutic effects of the premixes in each example and comparative example were evaluated through animal experiments. 300 one-day-old AA broilers were used in the experiment, and a negative control group, a positive control group, an antibiotic treatment group, and a traditional probiotic treatment group were also set up, with 5 replicates per group and 12 chickens per replicate, half male and half female.

[0081] In this study, the negative control group did not undergo proventriculitis modeling; the positive control group underwent proventriculitis modeling but did not receive any treatment; the antibiotic treatment group had 0.2 wt.% enrofloxacin added to the complete feed; and the traditional probiotic treatment group had 0.5 wt.% of a conventional compound bacteria (Bacillus subtilis + Lactobacillus acidophilus + Saccharomyces cerevisiae, in a mass ratio of 1:1:1) added to the complete feed. Premixed treatment groups corresponding to the examples and comparative examples were prepared by adding 1.2 wt.% of the compound to the complete feed.

[0082] The complete feed used in this experiment consists of the following components by weight percentage: corn 60%, soybean meal 30.4%, corn gluten meal 3%, soybean oil 2%, limestone powder 1.2%, dicalcium phosphate 1.8%, salt 0.36%, methionine 0.12%, lysine 0.12%, and multivitamin and mineral premix 1%.

[0083] Except for the negative control group, each treatment group was orally administered a suspension containing AFB1 (200 μg / kg) and avian adenovirus (105 TCID50) for 3 consecutive days starting from day 7 to establish a proventriculitis model. The modeling status was assessed at day 10, and each group was fed a treatment diet for 5 consecutive days. The growth performance of each group was recorded at day 15, and necropsy was performed to collect relevant indicators.

[0084] At 10 days of age, two chickens from each replicate in each group were necropsized to observe proventricular lesions. Serum and proventricular mucosa were collected to detect inflammatory factors, and the results are shown in Table 1. The results showed that the broilers in the negative control group exhibited normal behavior and normal proventricular morphology; while the broilers in other groups showed decreased feed intake and more severe proventricular swelling, manifested as white patches on the surface of the proventriculus, isthmic edema, and significantly higher levels of TNF-α in the mucosa and serum IL-6 than in the negative control group.

[0085] Table 1

[0086] Mucosal TNF-α (ng / mg) Serum IL-6 (pg / mL) Negative control group 0.33±0.05 16.22±1.69 Positive control group 3.61±0.33 69.26±3.64 Antibiotic prophylaxis group 3.84±0.16 58.43±5.88 Traditional probiotic prophylaxis group 3.69±0.24 61.75±8.23 Example / Comparative Example prophylaxis group 3.87±0.29 64.64±4.59

[0087] Clinical manifestations of broilers in each group were observed at 15 days of age, growth performance was statistically analyzed, and two chickens from each replicate of each group were necropsydid to observe proventricular lesions. Serum and proventricular mucosa were collected to detect inflammatory factors, and aflatoxin B1 residues in the proventricular stomach were detected and recorded in Table 2.

[0088] Table 2

[0089]

[0090]

[0091] The results showed that the positive control group broilers continued to show a decrease in feed intake and reduced activity. The feed intake of the premix treatment group, antibiotic treatment group and traditional probiotic treatment group in each comparison ratio increased. The feed intake of the premix treatment group in each example increased significantly. At 15 days of age, the average weight of the premix treatment group was close to that of the negative control group.

[0092] Necropsy revealed significant improvement in proventricular lesions in the premix treatment groups of each embodiment. The levels of TNF-α in the proventricular mucosa and serum IL-6 were significantly reduced, and the residual AFB1 in the proventriculus was also significantly lower than in the positive control group, antibiotic treatment group, traditional probiotic treatment group, and comparative premix treatment group. These results indicate that the premix prepared according to this invention, based on the interaction between bacteria and enzymes to prevent and treat proventricular inflammation in broilers, has excellent therapeutic effects.

[0093] Although the present invention has been illustrated and described with specific embodiments, it should be understood that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; those skilled in the art should understand that modifications can be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein, without departing from the spirit and scope of the present invention; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention; therefore, this means that all such substitutions and modifications that fall within the scope of the present invention are included in the appended claims.

Claims

1. A premix for preventing proventriculitis in chickens, characterized in that, The premix comprises the following components by weight: 120-180 parts of compound probiotic microcapsules, 60-100 parts of compound enzyme preparation, 250-330 parts of functional excipients, and 390-570 parts of carrier. The functional excipients include the following components by weight: 18-25 parts of nano-modified aluminosilicate, 4-7 parts of Jerusalem artichoke powder, 2.5-4 parts of curcumin microcapsules, and 0.8-1.5 parts of vitamin E succinate. The compound probiotic microcapsules are prepared by drying a W / O / W emulsion, wherein the W / O / W emulsion comprises an outer aqueous phase, an intermediate oil phase, and an inner aqueous phase, and the inner aqueous phase comprises a probiotic compound agent. The probiotic complex includes Bacillus subtilis, Saccharomyces boulardii, and Pediococcus lactis; the complex enzyme preparation includes aflatoxin-degrading enzyme, acidic protease, and β-glucanase; the nano-modified aluminosilicate is obtained by surface amylation modification of aluminosilicate.

2. The premix according to claim 1, characterized in that, The number of live bacteria in the compound probiotic microcapsules includes: Bacillus subtilis ≥ 5.0 * 10⁻⁶. 10 CFU / g, *Saccharomyces boulardii* ≥ 2.0*10 10 CFU / g, Pediococcus lactis ≥1.0*10 11 CFU / g.

3. The premix according to claim 1, characterized in that, The enzyme activity of the compound enzyme preparation includes: aflatoxin-degrading enzyme ≥15000U / g, acidic protease ≥25000U / g, and β-glucanase ≥18000U / g.

4. The premix according to claim 1, characterized in that, The particle size of the nano-modified aluminosilicate is 80nm~200nm.

5. The premix according to claim 1, characterized in that, The premix satisfies at least one of the following characteristics (a) to (d): (a) The volume ratio of the outer aqueous phase, the intermediate oil phase, and the inner aqueous phase is (8~12):(3~5):1; (b) The outer aqueous phase comprises an aqueous solution of chitosan and acetic acid, wherein the content of chitosan is 0.08 wt.% to 0.3 wt.%. (c) The intermediate oil phase comprises at least one of monoglycerides, polyglycerides, or lecithin; the intermediate oil phase further comprises one of beeswax or shellac; (d) The inner aqueous phase further includes at least one of trehalose or mannitol; the inner aqueous phase further includes at least one of glutamine or arginine.

6. The method for preparing the premix for preventing proventriculitis in chickens as described in any one of claims 1 to 5, characterized in that, Includes the following steps: The components of the intermediate oil phase and the inner aqueous phase are mixed and homogenized at 8000 rpm to 15000 rpm until homogeneous. Then, the outer aqueous phase is added and homogenized at 4000 rpm to obtain a W / O / W emulsion. The W / O / W emulsion is then spray-dried at low temperature to obtain composite probiotic microcapsules. The compound probiotic microcapsules, compound enzyme preparation, functional excipients and carrier are mixed through four-stage amplification to obtain a premix precursor. The premix precursor is then sealed and activated at 30℃~40℃ to obtain a premix for preventing and treating proventriculitis in chickens. The four-stage amplification and mixing process includes the following steps: S1. Thoroughly mix the nano-modified aluminosilicate, Jerusalem artichoke powder, and the carrier; S2. Thoroughly mix the mixture from S1 with the compound enzyme preparation; S3. Thoroughly mix the mixture from S2, the compound probiotic microcapsules, and the curcumin microcapsules; S4. Spray vitamin E succinate into the mixture in S3.

7. The preparation method according to claim 6, characterized in that, The inlet air temperature of the low-temperature spray dryer is 40℃~60℃, the outlet air temperature of the low-temperature spray dryer is 20℃~30℃, and the atomization pressure of the low-temperature spray dryer is 0.7MPa~0.85MPa.

8. An animal feed for preventing proventriculitis, characterized in that, Including the premix for preventing proventriculitis in chickens as described in any one of claims 1 to 5.

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