Proventriculitis molding agent as well as preparation method and application thereof

Abstract

The invention relates to the technical field of experimental animal pathological model modeling, in particular to a proventriculitis modeling agent and a preparation method and application thereof. The proventriculitis molding agent comprises a ZnO-OTA core and a mesoporous silica shell layer, wherein the surface of the ZnO-OTA core is coated with the mesoporous silica shell layer; the ZnO-OTA core is prepared from aminated zinc oxide nano particles and ochratoxin A; in the proventriculitis molding agent disclosed by the invention, after aminated zinc oxide nanoparticles and ochratoxin A are combined through an electrostatic adsorption effect, mesoporous silicon dioxide is adopted for coating to form a core-shell structure; the method is applied to construction of the broiler chicken proventriculitis pathological model, the stable proventriculitis model with the high pathological coincidence rate can be established, the required time is short, the modeling effect is good, and the stable proventriculitis model with the high pathological coincidence rate can be established within 21 days.

Description

Technical Field

[0001] The present invention relates to the technical field of experimental animal pathological model establishment, and particularly to a gizzard gastritis modeling agent, a preparation method thereof, and an application thereof. Background Art

[0002] Gizzard gastritis is a highly prevalent digestive system disease in broiler chicken farming. In recent years, with the expansion of intensive farming scale, its incidence rate has shown a significant upward trend, and it has become one of the important bottlenecks restricting the healthy development of the poultry industry. Gizzard gastritis mainly attacks the glandular stomach tissue of broiler chickens, resulting in inflammatory reactions in the mucosal layer, destruction of glandular structures, and digestive function disorders. The clinical manifestations include reduced appetite, growth stagnation, weight loss, fluffy and dull feathers, diarrhea, and a significant reduction in feed conversion rate. In severe cases, even a relatively high mortality rate may occur. Gizzard gastritis not only directly causes a decline in the production performance of broiler chickens but also reduces the body's immunity, secondary bacterial or viral infections (such as Escherichia coli, Salmonella, infectious bronchitis virus, etc.), forming a vicious cycle and exacerbating the economic losses of the farm. Therefore, constructing a reliable disease model to deeply analyze its pathogenesis and developing new prevention and control technologies based on this have become an urgent need in current research.

[0003] The construction of gizzard gastritis experimental models mainly relies on chemical induction methods, pathogenic microorganism infection methods, and feed factor induction methods. However, all current methods have significant limitations. Chemical induction methods usually use irritating substances such as strong acids, strong bases, or alcohol for gavage to simulate inflammatory damage by directly corroding the glandular stomach mucosa. This method is simple to operate and low in cost, but the modeling effect is quite different from natural disease occurrence. Pathogenic microorganism infection methods induce diseases by inoculating specific pathogens, which can partially simulate the natural infection process. The success rate of this modeling method is greatly affected by virus virulence, inoculation dose, and host immune status. Moreover, the requirements for virus amplification and preservation technology are high, and the experimental cycle is long, making it difficult to meet the needs of high-throughput drug screening. Feed factor induction methods simulate feed-derived gizzard gastritis by adding mycotoxins or high-salt / high-fiber feeds to the diet. Such methods can better reflect the chronic lesions caused by feed contamination in actual production, but the modeling cycle is long, and it is difficult to precisely control the toxin dose. Therefore, developing a highly efficient and stable gizzard gastritis modeling agent has become the key to breaking through the current research bottleneck.

[0004] In view of this, this invention is specifically proposed. Summary of the Invention

[0005] The first object of the present invention is to provide a gizzard gastritis modeling agent, which can establish a stable gizzard gastritis model with a high pathological compliance rate, requires a short time, and has a good modeling effect.

[0006] The second object of the present invention is to provide a preparation method of a gizzard gastritis modeling agent.

[0007] The third object of the present invention is to provide the application of the gizzard gastritis modeling agent in the preparation of products for constructing a pathological model of gizzard gastritis in broiler chickens.

[0008] In order to achieve the above objects of the present invention, the following technical solutions are specifically adopted:

[0009] In the first aspect, the present invention provides a gizzard gastritis modeling agent, which includes a ZnO-OTA core and a mesoporous silica shell layer coated on the surface of the ZnO-OTA core;

[0010] The ZnO-OTA core includes amino-functionalized zinc oxide nanoparticles and ochratoxin A.

[0011] Further, in the gizzard gastritis modeling agent, the content of the mesoporous silica shell layer is 60 wt% to 70 wt%.

[0012] Further, in the gizzard gastritis modeling agent, the content of the amino-functionalized zinc oxide nanoparticles is 29 wt% to 39.5 wt%, and the content of ochratoxin A is 0.5 wt% to 1 wt%.

[0013] In the second aspect, the present invention also provides a preparation method of the gizzard gastritis modeling agent as described above, including the following steps:

[0014] S1. Amino-functionalize zinc oxide nanoparticles with an amino modifier to obtain amino-functionalized zinc oxide nanoparticles;

[0015] S2. Mix the amino-functionalized zinc oxide nanoparticles and ochratoxin A to obtain a ZnO-OTA core;

[0016] S3. React the solution of the ZnO-OTA core, cetyltrimethylammonium bromide, ammonia water and tetraethyl orthosilicate to obtain the gizzard gastritis modeling agent.

[0017] Further, in step S1, at least one of the following features (1) to (4) is included;

[0018] (1) The amino modifier includes 3-aminopropyltriethoxysilane;

[0019] (2) The particle size of the zinc oxide nanoparticles is 50 to 80 nm;

[0020] (3) The dosage ratio of the zinc oxide nanoparticles to the amino modifier is 1 g: 1.9 to 2.1 mL;

[0021] (4) The amino-functionalization modification includes: reacting zinc oxide nanoparticles, an amino modifier and a solvent at 63 to 67 °C for 5 to 7 h to obtain the amino-functionalized zinc oxide nanoparticles.

[0022] Further, in step S2, the mass ratio of the amino-functionalized zinc oxide nanoparticles to the ochratoxin A is 4-6:1;

[0023] and / or, after mixing the amino-functionalized zinc oxide nanoparticles, the ochratoxin A and PBS buffer solution and stirring in the dark for 11-13 h, the ZnO-OTA core is obtained.

[0024] Further, in step S3, it includes at least one of the following features (1) to (3);

[0025] (1) The dosage ratio of the ZnO-OTA core in the ZnO-OTA core solution to the tetraethyl orthosilicate is 0.08-0.15 g:1.5 mL;

[0026] (2) The dosage ratio of the ZnO-OTA core, cetyltrimethylammonium bromide and ammonia water in the ZnO-OTA core solution is 0.08-0.15 g:0.3-0.5 g:1.5-3 mL;

[0027] (3) The temperature of the reaction is 34-36 °C, and the time of the reaction is 23-25 h.

[0028] In the third aspect, the present invention also provides the application of the adenomyogastritis modeling agent as described above in the preparation of a product for constructing a pathological model of broiler adenomyogastritis.

[0029] Further, the product for constructing a pathological model of broiler adenomyogastritis includes feed for inducing broiler adenomyogastritis.

[0030] Further, in the feed for inducing broiler adenomyogastritis, the content of the adenomyogastritis modeling agent is 3-5 mg / kg.

[0031] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0032] In the adenomyogastritis modeling agent of the present invention, after the amino-functionalized zinc oxide nanoparticles and ochratoxin A are combined by electrostatic adsorption, a core-shell structure is formed by coating with mesoporous silica; when it is applied to the construction of a pathological model of broiler adenomyogastritis, through the pH-responsive nanocarrier, the specific release of pathogenic factors in the glandular stomach area is realized, the OTA dose is controlled below 50% of the EU limit value, and the total amount of ZnO is lower than the broiler nutrition addition standard, with low systemic toxicity; a stable adenomyogastritis model with a high pathological coincidence rate can be established within 21 days, which is shortened by more than 30% compared with the traditional method (28-35 days). Detailed implementation manners

[0033] The technical solution of the present invention will be clearly and completely described below in conjunction with specific embodiments. However, those skilled in the art will understand that the following described embodiments are some embodiments of the present invention, rather than 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 of ordinary skill in the art without making creative efforts belong to the scope of protection of the present invention. For those conditions not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. Those reagents or instruments not specified by the manufacturer can be obtained as conventional products through commercial purchase.

[0034] The following specifically describes a gizzard gastritis modeling agent, its preparation method and application according to an embodiment of the present invention.

[0035] In some embodiments of the present invention, a gizzard gastritis modeling agent is provided, which includes a ZnO-OTA core and a mesoporous silica (mSiO) shell layer coated on the surface of the ZnO-OTA core;

[0036] The ZnO-OTA core includes amino-functionalized zinc oxide nanoparticles (ZnO-NH) and ochratoxin A (OTA).

[0037] In the gizzard gastritis modeling agent of the present invention, after the amino-functionalized zinc oxide nanoparticles and ochratoxin A are combined through electrostatic adsorption, ochratoxin A is loaded on the amino-functionalized zinc oxide nanoparticles, and then mesoporous silica is used for coating to form a core-shell structure; wherein, the mesoporous silica shell layer dissolves in the acidic environment (pH≈2.5) of the gizzard stomach, exposing the ZnO-OTA core, and the amino-functionalized zinc oxide nanoparticles generate Zn2 + , inhibiting the activity of mitochondrial complex III in gastric mucosal cells. At the same time, ochratoxin A improves the bioavailability through the nanocarrier, enhancing the inhibitory effect on ribosomal peptidyl transferase, thereby inducing gizzard gastritis.

[0038] The present invention applies mesoporous silica-coated ZnO-OTA to the construction of a pathological model of gizzard gastritis in broilers. Through a pH-responsive nanocarrier, the specific release of pathogenic factors in the gizzard stomach area is achieved, the OTA dose is controlled below 50% of the EU limit, and the total amount of ZnO is lower than the broiler nutrition addition standard, with low systemic toxicity; a stable gizzard gastritis model with a high pathological coincidence rate can be established within 21 days, which is shortened by more than 30% compared with the traditional method (28-35 days).

[0039] In some embodiments of the present invention, in the agent for inducing gizzard gastritis, the content of the mesoporous silica shell layer is 60 wt% to 70 wt%; typically but not restrictively, for example, in the agent for inducing gizzard gastritis, the content of the mesoporous silica shell layer can be 60 wt%, 62 wt%, 64 wt%, 66 wt%, 68 wt%, 70 wt% or a range value composed of any two of them.

[0040] In some embodiments of the present invention, in the agent for inducing gizzard gastritis, the content of the amino-functionalized zinc oxide nanoparticles is 29 wt% to 39.5 wt%, and the content of ochratoxin A is 0.5 wt% to 1 wt%; typically but not restrictively, for example, in the agent for inducing gizzard gastritis, the content of the amino-functionalized zinc oxide nanoparticles can be 29 wt%, 32 wt%, 34 wt%, 36 wt%, 38 wt%, 39.5 wt% or a range value composed of any two of them, and the content of ochratoxin A can be 0.5 wt%, 0.6 wt%, 0.7 wt%, 0.8 wt%, 0.9 wt%, 1 wt% or a range value composed of any two of them.

[0041] In the agent for inducing gizzard gastritis of the present invention, if the content of the amino-functionalized zinc oxide nanoparticles is too high, the toxicity will increase, and Zn2 + excessively inhibits mitochondrial respiratory chain complex IV, leading to metabolic acidosis; when the blood zinc concentration > 5 ppm, it inhibits the activity of NK cells; if the content of the amino-functionalized zinc oxide nanoparticles is too low, the mucosal damage is insufficient, and the amount of ROS generated is not enough to destroy the tight junction proteins; too low nanoparticle concentration leads to a decrease in the OTA loading rate.

[0042] In the agent for inducing gizzard gastritis of the present invention, if the content of ochratoxin A is too high, the acute toxicity and the mortality rate will increase; it inhibits the activity of systemic RNA polymerase III, resulting in non-specific damage; if the content of ochratoxin A is too low, the pathogenic effect is weak.

[0043] In the agent for inducing gizzard gastritis of the present invention, if the content of the mesoporous silica is too high, excessive SiO particles will cause mechanical wear of the gizzard; if the content of the mesoporous silica is too low, the stability is poor, and the degradation rate of OTA during storage increases.

[0044] In some embodiments of the present invention, a preparation method of the above agent for inducing gizzard gastritis is also provided, including the following steps:

[0045] S1. Amino-functionalize zinc oxide nanoparticles with an amino modifier to obtain amino-functionalized zinc oxide nanoparticles;

[0046] S2. Mix the amino-functionalized zinc oxide nanoparticles and ochratoxin A to obtain a ZnO-OTA core;

[0047] S3. React the solution of ZnO-OTA core, cetyltrimethylammonium bromide, ammonia water and tetraethyl orthosilicate to obtain the agent for inducing gizzard erosion.

[0048] In the preparation method of the agent for inducing gizzard erosion of the present invention, after the zinc oxide (ZnO) nanoparticles are aminated, they are combined with ochratoxin A (OTA) through electrostatic adsorption. Ochratoxin A is loaded on the aminated zinc oxide nanoparticles, and then a layer of mesoporous silica (mSiO₂) is coated to obtain the ZnO-OTA-mSiO₂ composite, that is, the agent for inducing gizzard erosion.

[0049] In some embodiments of the present invention, in step S1, the amino modifier includes 3-aminopropyltriethoxysilane.

[0050] In some embodiments of the present invention, in step S1, the particle size of the zinc oxide nanoparticles is 50-80 nm.

[0051] In some embodiments of the present invention, in step S1, the dosage ratio of the zinc oxide nanoparticles to the amino modifier is 1 g: 1.9-2.1 mL.

[0052] In some embodiments of the present invention, in step S1, the amination modification includes: reacting the zinc oxide nanoparticles, the amino modifier and the solvent at 63-67 °C for 5-7 h to obtain the aminated zinc oxide nanoparticles; preferably, the solvent includes anhydrous ethanol.

[0053] In some specific embodiments of the present invention, in step S1, zinc oxide nanoparticles (ZnO NPs) with a particle size of 50-80 nm are dispersed in anhydrous ethanol. After ultrasonic treatment, 3-aminopropyltriethoxysilane (APTES) is added, and the mixture is magnetically stirred and reacted at 63-67 °C for 5-7 h; then centrifuged to collect the precipitate; the precipitate is washed with anhydrous ethanol and vacuum dried to obtain the aminated nano-zinc oxide particles (ZnO-NH₂).

[0054] In some embodiments of the present invention, in step S2, the mass ratio of the aminated zinc oxide nanoparticles to ochratoxin A is 4-6: 1; typically but not limited to, for example, the mass ratio of the aminated zinc oxide nanoparticles to ochratoxin A can be 4: 1, 5: 1, 6: 1 or the range value between any two of them.

[0055] In some embodiments of the present invention, in step S2, the aminated zinc oxide nanoparticles, ochratoxin A and PBS buffer solution are mixed and stirred in the dark for 11-13 h to obtain the ZnO-OTA core.

[0056] In some embodiments of the present invention, in step S2, the dosage ratio of ochratoxin A to PBS buffer is 0.8 - 1 mg: 1 mL; preferably, the pH of the PBS buffer is 7.3 - 7.5.

[0057] In some specific embodiments of the present invention, in step S2, ochratoxin A (OTA) is dissolved in PBS buffer with a pH of 7.3 - 7.5; amino-functionalized zinc oxide nanoparticles are added and ultrasonically dispersed; stirring is carried out in the dark at 25 °C for 11 - 13 h, so that OTA is loaded on the amino-functionalized zinc oxide nanoparticles through electrostatic adsorption; then centrifugation is carried out to collect the precipitate; the precipitate is washed with PBS buffer to remove free OTA, and a ZnO-OTA core is obtained.

[0058] In some embodiments of the present invention, in step S3, the solution of the ZnO-OTA core includes an aqueous solution of the ZnO-OTA core.

[0059] In some embodiments of the present invention, in step S3, the dosage ratio of the ZnO-OTA core to tetraethyl orthosilicate in the solution of the ZnO-OTA core is 0.08 - 0.15 g: 1.5 mL.

[0060] In some embodiments of the present invention, in step S3, the dosage ratio of the ZnO-OTA core, cetyltrimethylammonium bromide, and ammonia water in the solution of the ZnO-OTA core is 0.08 - 0.15 g: 0.3 - 0.5 g: 1.5 - 3 mL.

[0061] In some embodiments of the present invention, in step S3, the reaction temperature is 34 - 36 °C, and the reaction time is 23 - 25 h.

[0062] In some specific embodiments of the present invention, in step S3, the ZnO-OTA core is dispersed in deionized water; cetyltrimethylammonium bromide (CTAB) is added and ultrasonically dispersed; ammonia water with a concentration of 25% - 28% is added and stirred; tetraethyl orthosilicate (TEOS) is added dropwise at a dropping rate of 0.08 - 0.12 mL / min, and stirring reaction is carried out at 34 - 36 °C for 23 - 25 h; then centrifugation is carried out to collect the precipitate; the precipitate is washed with ethanol to remove the CTAB template and vacuum dried to obtain an adenomyogastritis modeling agent (ZnO-OTA-mSiO composite).

[0063] Some embodiments of the present invention also provide the application of the above-mentioned adenomyogastritis modeling agent in the preparation of a product for constructing a pathological model of broiler adenomyogastritis.

[0064] In some embodiments of the present invention, the product for constructing a pathological model of broiler adenomyogastritis includes feed for inducing broiler adenomyogastritis.

[0065] In some embodiments of the present invention, in the feed for inducing gizzard gastritis in broiler chickens, the content of the gizzard gastritis modeling agent is 3-5 mg / kg.

[0066] In some embodiments of the present invention, the feed for inducing gizzard gastritis in broiler chickens further includes corn starch, and the mass ratio of the gizzard gastritis modeling agent to corn starch is 1:8-10. Corn starch is used as a carrier and a diluent.

[0067] In some embodiments of the present invention, the feed for inducing gizzard gastritis in broiler chickens further includes broiler chicken feed.

[0068] In some embodiments of the present invention, the method for preparing the feed for inducing gizzard gastritis in broiler chickens includes the following steps:

[0069] Mix the gizzard gastritis modeling agent and corn starch evenly, and mix them step by step to obtain a composite; then mix the composite and broiler chick feed step by step to the required amount to obtain the feed for inducing gizzard gastritis in broiler chickens.

[0070] In some embodiments of the present invention, a method for constructing a pathological model of gizzard gastritis in broiler chickens is also provided, including the following steps:

[0071] Feed the above-mentioned feed for inducing gizzard gastritis in broiler chickens to the broiler chickens; preferably, the feeding time ≤ 21 days; free feeding is sufficient.

[0072] Using the gizzard gastritis modeling agent of the present invention to construct a pathological model of gizzard gastritis in broiler chickens, based on the technology of "mycotoxin-nanocarrier complex" inducing gizzard gastritis in broiler chickens, a method for constructing an efficient, controllable and repeatable experimental model of gizzard gastritis in broiler chickens is provided, which requires a short time and has a good modeling effect.

[0073] Example 1

[0074] The method for preparing the gizzard gastritis modeling agent provided in this example includes the following steps:

[0075] S1. Disperse 1.0 g of zinc oxide nanoparticles (ZnO NPs) with a particle size of 50-80 nm in 200 mL of absolute ethanol. After ultrasonic treatment for 30 min (power 400 W, pulse mode), add 2.0 mL of 3-aminopropyltriethoxysilane (APTES), and magnetically stir and react at 65 °C for 6 h; then centrifuge at a speed of 12,000 rpm for 10 min to collect the precipitate; wash the precipitate 3 times with absolute ethanol and vacuum dry at 60 °C for 12 h to obtain amino-functionalized zinc oxide nanoparticles (ZnO-NH);

[0076] S2. Dissolve 50 mg of ochratoxin A (OTA) in 50 mL of PBS buffer with a pH of 7.4; add 200 mg of the above-mentioned amino-functionalized zinc oxide nanoparticles, and ultrasonically disperse for 15 min (power 300 W); stir in the dark at 25 °C for 12 h; then centrifuge at 12,000 rpm for 10 min to collect the precipitate; wash the precipitate twice with PBS buffer to obtain the ZnO-OTA core;

[0077] S3. Disperse 100 mg of the above ZnO-OTA core in 100 mL of deionized water; add 0.5 g of cetyltrimethylammonium bromide (CTAB), and ultrasonically disperse for 20 min (power 350 W); add 2 mL of ammonia water (concentration 25% - 28%), and stir for 15 min; dropwise add 1.5 mL of tetraethyl orthosilicate (TEOS) at a dropping rate of 0.1 mL / min, and stir and react at 35 °C for 24 h; then centrifuge at 12,000 rpm for 10 min to collect the precipitate; wash the precipitate three times with ethanol and vacuum dry at 60 °C for 12 h to obtain the gizzard-adenitis modeling agent (ZnO-OTA-mSiO composite).

[0078] The preparation method of the feed for inducing gizzard-adenitis in broilers provided by this example includes the following steps:

[0079] Mix 500 mg of the gizzard-adenitis modeling agent of this example evenly with 4500 mg of corn starch, and mix three times step by step to obtain 500 g of the composite; then mix the composite with broiler chick feed step by step to 100 kg, and the feed for inducing gizzard-adenitis in broilers is obtained.

[0080] Example 2

[0081] The preparation method of the gizzard-adenitis modeling agent provided by this example includes the following steps:

[0082] S1. Disperse 1.0 g of zinc oxide nanoparticles (ZnO NPs) with a particle size of 50 - 80 nm in 200 mL of absolute ethanol. After ultrasonic treatment for 30 min (power 400 W, pulse mode), add 2.1 mL of 3-aminopropyltriethoxysilane (APTES), and magnetically stir and react at 67 °C for 7 h; then centrifuge at 12,000 rpm for 10 min to collect the precipitate; wash the precipitate three times with absolute ethanol and vacuum dry at 60 °C for 12 h to obtain amino-functionalized zinc oxide nanoparticles (ZnO-NH);

[0083] S2. Dissolve 50 mg of ochratoxin A (OTA) in 50 mL of PBS buffer with a pH of 7.45; add 300 mg of the above-mentioned amino-functionalized zinc oxide nanoparticles, and ultrasonically disperse for 30 min (power 300 W); stir in the dark at 25 °C for 13 h; then centrifuge at 12,000 rpm for 10 min to collect the precipitate; wash the precipitate twice with PBS buffer to obtain the ZnO-OTA core;

[0084] S3. Disperse 120 mg of the above ZnO-OTA core in 100 mL of deionized water; add 0.4 g of cetyltrimethylammonium bromide (CTAB), and ultrasonically disperse for 20 min (power 350 W); add 3 mL of ammonia water (concentration 25% - 28%), and stir for 15 min; dropwise add 1.5 mL of tetraethyl orthosilicate (TEOS) at a dropping rate of 0.12 mL / min, and stir and react at 36 °C for 25 h; then centrifuge at 12,000 rpm for 10 min to collect the precipitate; wash the precipitate three times with ethanol and vacuum dry at 60 °C for 12 h to obtain the gizzard-adenitis modeling agent (ZnO-OTA-mSiO composite).

[0085] The preparation method of the feed for inducing gizzard-adenitis in broilers provided in this example includes the following steps:

[0086] Mix 500 mg of the gizzard-adenitis modeling agent of this example evenly with 4500 mg of corn starch, and mix three times step by step to obtain 500 g of the composite; then mix the composite with broiler starter feed step by step to 100 kg to obtain the feed for inducing gizzard-adenitis in broilers.

[0087] Example 3

[0088] The preparation method of the gizzard-adenitis modeling agent provided in this example includes the following steps:

[0089] S1. Disperse 1.0 g of zinc oxide nanoparticles (ZnO NPs) with a particle size of 50 - 80 nm in 200 mL of absolute ethanol. After ultrasonic treatment for 30 min (power 400 W, pulse mode), add 2.0 mL of 3-aminopropyltriethoxysilane (APTES), and magnetically stir and react at 65 °C for 6 h; then centrifuge at 12,000 rpm for 10 min to collect the precipitate; wash the precipitate three times with absolute ethanol and vacuum dry at 60 °C for 12 h to obtain amino-functionalized zinc oxide nanoparticles (ZnO-NH);

[0090] S2. Dissolve 50 mg of ochratoxin A (OTA) in 50 mL of PBS buffer with a pH of 7.4; add 200 mg of the above-mentioned amino-functionalized nano-zinc oxide particles, and ultrasonically disperse for 15 min (power 300 W); stir in the dark at 25 °C for 12 h; then centrifuge at a speed of 12,000 rpm for 10 min to collect the precipitate; wash the precipitate twice with PBS buffer to obtain the ZnO-OTA core;

[0091] S3. Disperse 80 mg of the above-mentioned ZnO-OTA core in 100 mL of deionized water; add 0.5 g of cetyltrimethylammonium bromide (CTAB), and ultrasonically disperse for 20 min (power 350 W); add 3 mL of ammonia water (concentration 25% - 28%), and stir for 15 min; dropwise add 1.5 mL of tetraethyl orthosilicate (TEOS) at a dropping rate of 0.1 mL / min, and stir and react at 36 °C for 12 h; then centrifuge at a speed of 12,000 rpm for 10 min to collect the precipitate; wash the precipitate three times with ethanol, and vacuum dry at 60 °C for 12 h to obtain the agent for inducing gizzard gastritis in broilers (ZnO-OTA-mSiO composite).

[0092] The preparation method of the feed for inducing gizzard gastritis in broilers provided by this example includes the following steps:

[0093] Mix 500 mg of the agent for inducing gizzard gastritis in broilers of this example with 4,500 mg of corn starch evenly, and mix three times step by step to obtain 500 g of the composite; then mix the composite with broiler starter feed step by step to 100 kg, and the feed for inducing gizzard gastritis in broilers is obtained.

[0094] Comparative Example 1

[0095] The agent for inducing gizzard gastritis provided by this comparative example is deoxynivalenol (DON).

[0096] The preparation method of the feed for inducing gizzard gastritis in broilers provided by this comparative example includes the following steps:

[0097] Mix 500 mg of deoxynivalenol (DON) with 4,500 mg of corn starch evenly, and mix three times step by step to obtain 500 g of the composite; then mix the composite with broiler starter feed step by step to 100 kg, and the feed for inducing gizzard gastritis in broilers is obtained.

[0098] Test Example 1

[0099] Select 1-day-old broilers, and divide them into a control group, model group A, model group B, model group C, and model group D, with 5 replicates in each group and 12 chickens in each replicate. Among them, the control group is fed with broiler starter feed, and model groups A - D are fed with the feeds for inducing gizzard gastritis in broilers of Example 1, Example 2, Example 3, and Comparative Example 1 respectively; continuously feed (free access to food) for 21 days.

[0100] During the experiment, the growth performance and clinical symptoms of broilers in each group were observed and recorded. After the experiment, 10 chickens were selected from each group to collect glandular stomach mucosa, and the contents of IL-6, TNF-α and IL-1β were detected by qPCR. The results are shown in Table 1 and Table 2.

[0101] In Table 1, the feed-to-gain ratio is: total feed intake / total weight gain.

[0102] Table 1

[0103] Average weight at 21 days of age (g) Average daily weight gain (g / d) Feed to meat ratio Control group 735.34±20.77 34.19±2.56 1.51±0.03 Model group A <![CDATA[618.28±26.21 * > <![CDATA[28.56±2.07 * > <![CDATA[1.62±0.01 * > Model group B 631.56±16.68 29.02±1.35 1.62±0.02 Model group C 672.77±18.14 31.21±1.24 1.58±0.01 Model group D 681.64±23.42 31.56±1.57 1.56±0.02

[0104] In Table 1, * indicates that there is a significant difference between model group A and the control group, P < 0.05.

[0105] As can be seen from Table 1, the broilers in model group A of glandular gastritis were listless, with reduced activity, fluffy and dull feathers, undigested particles in the feces, and severe loose stools.

[0106] The average weight and average daily weight gain of 21-day-old broilers in model group A of glandular gastritis were significantly lower than those of the control group, and the feed-to-gain ratio was significantly higher than that of the control group.

[0107] Table 2

[0108] IL-6 (pg / mL) TNF-α (pg / mL) IFN-γ (pg / mL) Control group 26.43±0.44 37.68±0.78 89.73±3.22 Model group A <![CDATA[48.78±0.97 * > <![CDATA[87.35±2.71 * > <![CDATA[194.39±2.64 * > Model group B 46.26±1.15 92.34±2.34 177.53±1.98 Model group C 37.74±0.38 61.29±1.89 143.26±2.95 Model group D 35.16±1.51 53.26±0.76 126.58±1.33

[0109] In Table 2, * indicates that there is a significant difference between model group A and the control group, P < 0.05.

[0110] As can be seen from Table 2, by detecting the contents of IL-6, TNF-α and IL-1β in the glandular stomach mucosa of each group by qPCR, the levels of inflammatory factors in the glandular stomach mucosa of model group A were significantly higher than those of the control group.

[0111] Using the glandular gastritis modeling agent of the present invention to construct a pathological model of glandular gastritis in broilers has a good modeling effect.

[0112] Finally, it should be noted that: the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements on some or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A gizzard erosion modeling agent, characterized in that, It includes a ZnO-OTA core and a mesoporous silica shell layer coated on the surface of the ZnO-OTA core; The ZnO-OTA core includes amino-functionalized zinc oxide nanoparticles and ochratoxin A.

2. The gizzard gastritis modeling agent according to claim 1, wherein In the adenomyogastritis modeling agent, the content of the mesoporous silica shell layer is 60wt% - 70wt%.

3. The gizzard gastritis modeling agent according to claim 1, wherein In the adenomyogastritis modeling agent, the content of the amino-functionalized zinc oxide nanoparticles is 29wt% - 39.5wt%, and the content of ochratoxin A is 0.5wt% - 1wt%.

4. The preparation method of the gizzard mycosis modeling agent according to any one of claims 1 to 3, characterized in that, It includes the following steps: S1. Amino-functionalize zinc oxide nanoparticles with an amino modifier to obtain amino-functionalized zinc oxide nanoparticles; S2. Mix the amino-functionalized zinc oxide nanoparticles and ochratoxin A to obtain a ZnO-OTA core; S3. React the solution of the ZnO-OTA core, cetyltrimethylammonium bromide, ammonia water and tetraethyl orthosilicate to obtain the adenomyogastritis modeling agent.

5. The preparation method of the gizzard gastritis modeling agent according to claim 4, characterized in that, In step S1, it includes at least one of the following features (1) to (4); (1) The amino modifier includes 3-aminopropyltriethoxysilane; (2) The particle size of the zinc oxide nanoparticles is 50 - 80nm; (3) The dosage ratio of the zinc oxide nanoparticles to the amino modifier is 1g: 1.9 - 2.1mL; (4) The amino-functionalization modification includes: reacting zinc oxide nanoparticles, an amino modifier and a solvent at 63 - 67°C for 5 - 7h to obtain the amino-functionalized zinc oxide nanoparticles.

6. The preparation method of the gizzard mycosis modeling agent according to claim 4, wherein, In step S2, the mass ratio of the amino-functionalized zinc oxide nanoparticles to the ochratoxin A is 4 - 6: 1; And / or, mix the amino-functionalized zinc oxide nanoparticles, the ochratoxin A and PBS buffer solution and stir in the dark for 11 - 13h to obtain the ZnO-OTA core.

7. The preparation method of the gizzard gastritis modeling agent according to claim 4, wherein, In step S3, it includes at least one of the following features (1) to (3); (1) The dosage ratio of the ZnO-OTA core in the solution of the ZnO-OTA core to the tetraethyl orthosilicate is 0.08 - 0.15g: 1.5mL; (2) The dosage ratio of the ZnO-OTA core in the solution of the ZnO-OTA core, the cetyltrimethylammonium bromide and the ammonia water is 0.08 - 0.15g: 0.3 - 0.5g: 1.5 - 3mL; (3) The reaction temperature is 34 - 36°C, and the reaction time is 23 - 25h.

8. The application of the adenomyogastritis modeling agent according to any one of claims 1 - 3 in the preparation of a product for constructing a pathological model of broiler adenomyogastritis.

9. The application according to claim 8, wherein The product for constructing a pathological model of broiler adenomyogastritis includes feed for inducing broiler adenomyogastritis.

10. The application according to claim 9, characterized in that, In the feed for inducing broiler adenomyogastritis, the content of the adenomyogastritis modeling agent is 3 - 5mg / kg.