Preparation for salpingitis of laying hens and preparation method thereof

Abstract

The invention is suitable for the technical field of veterinary drugs, and provides a preparation for salpingitis of laying hens and a preparation method thereof.The preparation is prepared from thyme essential oil, modified resveratrol, modified chitosan, sodium alginate-zinc ion gel microspheres, modified epigallocatechin gallate and mussel mucin. According to the invention, the sodium alginate-zinc ion gel microspheres are intelligently swelled in an acidic inflammation environment of the fallopian tube, and zinc ions are released to inhibit a pathogenic bacteria biological membrane; the zinc oxide nanorods directionally arranged in the magnetic field enhance the tissue permeability and improve the drug concentration of the focus; mussel mucoprotein is covalently crosslinked with tissue protein through catechol groups, so that the residence time is prolonged; activated amino groups of the modified chitosan are combined with negative charges of mucous membranes to form a physical barrier to block pathogen adsorption; the thyme essential oil destroys bacterial cell membranes, so that the permeation efficiency of subsequent antibacterial components is improved; the free radical scavenging capacity of the modified resveratrol-quercetin eutectic is improved compared with that of a monomer.

Description

Technical Field

[0001] The invention belongs to the technical field of veterinary medicines, and in particular relates to a preparation for laying hen salpingitis and a preparation method thereof. Background Art

[0002] Salpingitis in laying hens is a common disease in intensive poultry farming, primarily caused by infection with pathogens such as Escherichia coli and Salmonella. Current prevention and treatment options primarily rely on antibiotics (such as enrofloxacin and doxycycline), but these are associated with issues such as drug residues, increased drug resistance, and contraindications during egg production. Natural plant extracts (such as essential oils and flavonoids), while relatively safe, have the following drawbacks: Low bioavailability: Resveratrol, epigallocatechin gallate (EGCG) and other ingredients are poorly water-soluble, with intestinal absorption rates of less than 10%; Weak targeting: Conventional preparations are difficult to accumulate at the site of inflammation, and the effective concentration is maintained for a short time. Summary of the Invention

[0003] The present invention provides a preparation for laying hen salpingitis, aiming to solve the above problems.

[0004] The present invention is achieved by providing a preparation for laying hen salpingitis, comprising the following raw materials in parts by weight: 5-8 parts of thyme essential oil, 3-6 parts of modified resveratrol, 8-12 parts of modified chitosan, 10-15 parts of sodium alginate-zinc ion gel microspheres, 4-7 parts of modified epigallocatechin gallate, and 5-10 parts of mussel mucin.

[0005] Preferably, the following raw materials are included in parts by weight: 6-7 parts of thyme essential oil, 4-5 parts of modified resveratrol, 9-11 parts of modified chitosan, 11-14 parts of sodium alginate-zinc ion gel microspheres, 5-6 parts of modified epigallocatechin gallate, and 6-9 parts of mussel mucin.

[0006] Preferably, the following raw materials are included in parts by weight: 6.5 parts of thyme essential oil, 4.5 parts of modified resveratrol, 10 parts of modified chitosan, 12.5 parts of sodium alginate-zinc ion gel microspheres, 5.5 parts of modified epigallocatechin gallate, and 7.5 parts of mussel mucin.

[0007] Preferably, the preparation method of the modified resveratrol is as follows: Resveratrol and quercetin were added to a ball mill at a molar ratio of 1:1.1-1.3, and 5-15 wt% anhydrous calcium sulfate was added; Under nitrogen protection, ball mill at 400-600 rpm for 3-5 hours, controlling the temperature ≤40°C; The eutectic powder was placed in an argon / oxygen (8-9:1) low-temperature plasma for 2-4 minutes (power 40-60W); The template is eluted with supercritical CO2 (pressure 20-30 MPa, temperature 35-45°C).

[0008] By forming a hydrogen bond co-crystal between resveratrol and quercetin, the solubility is improved; argon-oxygen plasma etches micropores on the co-crystal surface, expanding the specific surface area and increasing the drug loading capacity; supercritical CO2 elutes the template to form a mesoporous structure, accelerating drug dissolution. The co-crystal inhibits NF-κB inflammatory signaling by activating the Nrf2 / ARE pathway.

[0009] Preferably, the preparation method of the modified chitosan is as follows: Dissolve chitosan with a deacetylation degree of ≥95% in 0.5-1.5% formic acid solution to prepare a 2-4wt% solution; Add sodium tripolyphosphate (the molar ratio of chitosan: sodium tripolyphosphate is 1:0.1-0.3) and stir at 45-55°C for 30-50 minutes to activate the amino group.

[0010] Chitosan was protonated by 0.5% formic acid to enhance electrostatic adsorption with bacterial membranes; sodium tripolyphosphate was cross-linked to form a quaternary ammonium salt structure, which reduced the minimum inhibitory concentration for Escherichia coli and activated the amino group to form a Schiff base bond with the catechol group of mussel mucin, thereby improving the mucosal adhesion strength.

[0011] Preferably, the preparation method of the sodium alginate-zinc ion gel microspheres is as follows: Internal phase: sodium alginate solution (2-4%) and zinc oxide nanorods (aspect ratio 15-25:1, 3-7%); External phase: soybean oil containing 1-3% sorbitan oleate and 0.3-0.7% ferroferric oxide nanoparticles (8-12 nm); dripping through a coaxial microfluidic chip (inner phase flow rate 0.4-0.6 mL / h, outer phase flow rate 4-6 mL / h); The droplets are oriented and aligned in a rotating magnetic field (1500-2500 Gs, 80-120 rpm), forming a vertical channel structure to increase drug loading; Cross-link and cure in 1-3% calcium chloride solution for 20-40 minutes.

[0012] Zinc oxide nanorods aligned in a magnetic field enhance tissue permeability and increase drug concentration in lesions. Sodium alginate-zinc ion gel microspheres intelligently swell in the acidic inflammatory environment of the fallopian tube (pH≈6.2), releasing zinc ions to inhibit pathogenic biofilms, promote epithelial cell proliferation, and repair the mucosa.

[0013] Preferably, the preparation method of the modified epigallocatechin gallate is as follows: Epigallocatechin gallate, L-arginine, and zinc chloride are reacted in a pH 7.2-7.6 buffer solution at a molar ratio of 1:0.9-1.1:0.4-0.6; Salmonella lipopolysaccharide (template molecule, 0.5-1.5% by mass) was added, and ethyl orthosilicate was added dropwise for sol-gel embedding; The template was eluted with a 0.05-0.15 M sodium citrate / ethylenediaminetetraacetic acid mixture; Modified thiol polyethylene glycol (SH-PEG, molecular weight 2000-5000Da, grafting amount 10-20%).

[0014] Through the ternary complexation of zinc ion, arginine and EGCG, the zinc ion is coordinated with the phenolic hydroxyl group of EGCG, and the arginine carboxyl group participates in the formation of an octahedral structure. Using Salmonella lipopolysaccharide as a template, a specific recognition cavity is formed to target and capture Salmonella. Thiol polyethylene glycol blocks the oxidation site to form a three-dimensional protective layer, avoiding clearance by the reticuloendothelial system and prolonging the plasma half-life.

[0015] The present invention also provides a method for preparing a preparation for laying hen salpingitis, comprising the following steps: Prepare the raw materials according to the ratio; Thyme essential oil, modified resveratrol, and modified epigallocatechin gallate were mixed and subjected to ultrasonic emulsification (power 300-500W, time 10-30min); Add modified chitosan, sodium alginate-zinc ion gel microspheres, and mussel mucin, and stir to mix at 4-6°C; The mixture was treated with a pulsed electric field (field strength 5-8 kV / cm, time 10-20 ms).

[0016] Preferably, after the mixture is treated with a pulsed electric field, it is frozen at -50 to -40°C for 2-4 hours, then thawed at 25 to 30°C for 1 hour, and the cycle is repeated 2-4 times to form a porous network structure. The pores are expanded through the ice crystal growth-dissolution effect, the porosity of the microspheres is increased, and the drug release at the inflammatory site is accelerated.

[0017] Compared with the prior art, the embodiments of the present application have the following beneficial effects: The preparation for laying hen salpingitis provided by the present invention comprises sodium alginate-zinc ion gel microspheres that intelligently swell in the acidic inflammatory environment of the oviduct, releasing zinc ions to inhibit pathogenic bacterial biofilms; zinc oxide nanorods arranged in a magnetic field enhance tissue permeability and increase drug concentration in lesions; mussel mucin is covalently cross-linked with tissue proteins via catechol groups, extending retention time and achieving a long-lasting effect; the activated amino groups of modified chitosan combine with the negative charge of the mucosa to form a physical barrier to block pathogen adsorption; thyme essential oil destroys bacterial cell membranes, thereby improving the penetration efficiency of subsequent antibacterial ingredients; and the free radical scavenging ability of the modified resveratrol-quercetin co-crystal is improved compared to that of the monomer. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a flow chart of a method for preparing a preparation for laying hen salpingitis provided in Examples 1-5 of the present invention.

[0019] Figure 2 This is a flow chart of a preparation method for laying hen salpingitis provided in Examples 6-8 of the present invention. DETAILED DESCRIPTION

[0020] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as commonly understood by those skilled in the art to which this application belongs. The terms used in the specification of the application are only for the purpose of describing specific embodiments and are not intended to limit this application. The terms "including" and "having" and any variations thereof in the specification and claims of this application and the above-mentioned drawings are intended to cover non-exclusive inclusions. The terms "first", "second", etc. in the specification and claims of this application or the above-mentioned drawings are used to distinguish different objects, not to describe a specific order.

[0021] References herein to "embodiments" mean that a particular feature, structure, or characteristic described in connection with the embodiments may be included in at least one embodiment of the present application. The appearance of this phrase in various places in the specification does not necessarily refer to the same embodiment, nor does it constitute an independent or alternative embodiment that is mutually exclusive of other embodiments. It is understood, both explicitly and implicitly, by those skilled in the art that the embodiments described herein may be combined with other embodiments.

[0022] Example 1 The embodiment of the present invention provides a preparation for laying hen salpingitis, comprising the following raw materials in parts by weight: 5 parts of thyme essential oil, 3 parts of modified resveratrol, 8 parts of modified chitosan, 10 parts of sodium alginate zinc ion gel microspheres, 4 parts of modified epigallocatechin gallate, and 5 parts of mussel mucin. The preparation method of the preparation for laying hen salpingitis is as follows: Figure 1 As shown, the following steps are included: Prepare the raw materials according to the ratio; Thyme essential oil, modified resveratrol, and modified epigallocatechin gallate were mixed and subjected to ultrasonic emulsification (power 300 W, time 10 min); Add modified chitosan, sodium alginate-zinc ion gel microspheres, and mussel mucin and stir to mix at 4°C; The mixture was treated with a pulsed electric field (field strength 5 kV / cm, time 10 ms).

[0023] Wherein, the preparation method of the modified resveratrol is as follows: Resveratrol and quercetin were added to a ball mill at a molar ratio of 1:1.1, and 5 wt% anhydrous calcium sulfate was added; Under nitrogen protection, ball mill at 400 rpm for 3 h, controlling the temperature ≤ 40 °C; The eutectic powder was placed in an argon / oxygen (8:1) low-temperature plasma for 2 min (power 40 W); The template was eluted with supercritical CO2 (pressure 20 MPa, temperature 35°C).

[0024] Furthermore, the preparation method of the modified chitosan is as follows: Chitosan with a deacetylation degree of ≥95% was dissolved in 0.5% formic acid solution to prepare a 2 wt% solution; Sodium tripolyphosphate (the molar ratio of chitosan: sodium tripolyphosphate is 1:0.1) was added and stirred at 45 °C for 30 min to activate the amino groups.

[0025] Furthermore, the preparation method of the sodium alginate-zinc ion gel microspheres is as follows: Internal phase: sodium alginate solution (2%) and zinc oxide nanorods (aspect ratio 15:1, 3%); External phase: soybean oil containing 1% sorbitan oleate and 0.3% ferrosoferric oxide nanoparticles (8 nm); dripping through a coaxial microfluidic chip (inner phase flow rate 0.4 mL / h, outer phase flow rate 4 mL / h); The droplets are oriented and aligned in a rotating magnetic field (1500 Gs, 80 rpm) to form a vertical channel structure, thereby increasing the drug loading capacity. Cross-link and cure in 1% calcium chloride solution for 20 minutes.

[0026] In this embodiment, the preparation method of the modified epigallocatechin gallate is as follows: Epigallocatechin gallate, L-arginine, and zinc chloride were reacted in a pH 7.2 buffer solution at a molar ratio of 1:0.9:0.4; Salmonella lipopolysaccharide (template molecule, 0.5% by mass) was added, and ethyl orthosilicate was added dropwise for sol-gel embedding; The template was eluted with a 0.05 M sodium citrate / ethylenediaminetetraacetic acid mixture; Modified thiol polyethylene glycol (SHPEG, molecular weight 2000Da, grafting amount 10%).

[0027] Example 2 The embodiment of the present invention provides a preparation for laying hen salpingitis, comprising the following raw materials in parts by weight: 6 parts of thyme essential oil, 4 parts of modified resveratrol, 9 parts of modified chitosan, 11 parts of sodium alginate zinc ion gel microspheres, 5 parts of modified epigallocatechin gallate, and 6 parts of mussel mucin. The preparation method of the preparation for laying hen salpingitis is as follows: Figure 1 As shown, the following steps are included: Prepare the raw materials according to the ratio; Thyme essential oil, modified resveratrol, and modified epigallocatechin gallate were mixed and subjected to ultrasonic emulsification (power 300 W, time 10 min); Add modified chitosan, sodium alginate-zinc ion gel microspheres, and mussel mucin and stir to mix at 4°C; The mixture was treated with a pulsed electric field (field strength 5 kV / cm, time 10 ms).

[0028] Wherein, the preparation method of the modified resveratrol is as follows: Resveratrol and quercetin were added to a ball mill at a molar ratio of 1:1.1, and 5 wt% anhydrous calcium sulfate was added; Under nitrogen protection, ball mill at 400 rpm for 3 h, controlling the temperature ≤ 40 °C; The eutectic powder was placed in an argon / oxygen (8:1) low-temperature plasma for 2 min (power 40 W); The template was eluted with supercritical CO2 (pressure 20 MPa, temperature 35°C).

[0029] Furthermore, the preparation method of the modified chitosan is as follows: Chitosan with a deacetylation degree of ≥95% was dissolved in 0.5% formic acid solution to prepare a 2 wt% solution; Sodium tripolyphosphate (the molar ratio of chitosan: sodium tripolyphosphate is 1:0.1) was added and stirred at 45 °C for 30 min to activate the amino groups.

[0030] Furthermore, the preparation method of the sodium alginate-zinc ion gel microspheres is as follows: Internal phase: sodium alginate solution (2%) and zinc oxide nanorods (aspect ratio 15:1, 3%); External phase: soybean oil containing 1% sorbitan oleate and 0.3% ferrosoferric oxide nanoparticles (8 nm); dripping through a coaxial microfluidic chip (inner phase flow rate 0.4 mL / h, outer phase flow rate 4 mL / h); The droplets are oriented and aligned in a rotating magnetic field (1500 Gs, 80 rpm) to form a vertical channel structure, thereby increasing the drug loading capacity. Cross-link and cure in 1% calcium chloride solution for 20 minutes.

[0031] In this embodiment, the preparation method of the modified epigallocatechin gallate is as follows: Epigallocatechin gallate, L-arginine, and zinc chloride were reacted in a pH 7.2 buffer solution at a molar ratio of 1:0.9:0.4; Salmonella lipopolysaccharide (template molecule, 0.5% by mass) was added, and ethyl orthosilicate was added dropwise for sol-gel embedding; The template was eluted with a 0.05 M sodium citrate / ethylenediaminetetraacetic acid mixture; Modified thiol polyethylene glycol (SHPEG, molecular weight 2000Da, grafting amount 10%).

[0032] Example 3 The embodiment of the present invention provides a preparation for laying hen salpingitis, comprising the following raw materials in parts by weight: 6.5 parts of thyme essential oil, 4.5 parts of modified resveratrol, 10 parts of modified chitosan, 12.5 parts of sodium alginate-zinc ion gel microspheres, 5.5 parts of modified epigallocatechin gallate, and 7.5 parts of mussel mucin. The preparation method of the preparation for laying hen salpingitis is as follows: Figure 1 As shown, the following steps are included: Prepare the raw materials according to the ratio; Thyme essential oil, modified resveratrol, and modified epigallocatechin gallate were mixed and subjected to ultrasonic emulsification (power 400 W, time 20 min); Add modified chitosan, sodium alginate-zinc ion gel microspheres, and mussel mucin, and stir to mix at 5°C; The mixture was treated with a pulsed electric field (field strength 6.5 kV / cm, time 15 ms).

[0033] Wherein, the preparation method of the modified resveratrol is as follows: Resveratrol and quercetin were added to a ball mill at a molar ratio of 1:1.2, and 10 wt% anhydrous calcium sulfate was added; Under nitrogen protection, ball milling was performed at 500 rpm for 4 h, and the temperature was controlled at ≤40°C; The eutectic powder was placed in an argon / oxygen (8.5:1) low-temperature plasma for 3 min (power 50 W); The template was eluted with supercritical CO2 (pressure 25 MPa, temperature 40°C).

[0034] Furthermore, the preparation method of the modified chitosan is as follows: Chitosan with a deacetylation degree of ≥95% was dissolved in 1% formic acid solution to prepare a 3 wt% solution; Sodium tripolyphosphate (the molar ratio of chitosan: sodium tripolyphosphate is 1:0.2) was added and stirred at 50°C for 40 min to activate the amino groups.

[0035] Furthermore, the preparation method of the sodium alginate-zinc ion gel microspheres is as follows: Internal phase: sodium alginate solution (3%) and zinc oxide nanorods (aspect ratio 20, 5%); External phase: soybean oil containing 2% sorbitan oleate and 0.5% ferrosoferric oxide nanoparticles (10 nm); dripping through a coaxial microfluidic chip (inner phase flow rate 0.5 mL / h, outer phase flow rate 5 mL / h); The droplets were oriented and aligned by a rotating magnetic field (2000 Gs, 100 rpm), forming a vertical channel structure to increase drug loading; Cross-link and cure in 2% calcium chloride solution for 30 minutes.

[0036] In this embodiment, the preparation method of the modified epigallocatechin gallate is as follows: Epigallocatechin gallate, L-arginine, and zinc chloride were reacted in a pH 7.4 buffer solution at a molar ratio of 1:1:0.5; Salmonella lipopolysaccharide (template molecule, 1% by mass) was added, and ethyl orthosilicate was added dropwise for sol-gel embedding; The template was eluted with a 0.1 M sodium citrate / ethylenediaminetetraacetic acid mixture; Modified thiol polyethylene glycol (SH-PEG, molecular weight 3500Da, grafting amount 15%).

[0037] Example 4 The embodiment of the present invention provides a preparation for laying hen salpingitis, comprising the following raw materials in parts by weight: 7 parts of thyme essential oil, 5 parts of modified resveratrol, 11 parts of modified chitosan, 14 parts of sodium alginate zinc ion gel microspheres, 6 parts of modified epigallocatechin gallate, and 9 parts of mussel mucin. The preparation method of the preparation for laying hen salpingitis is as follows: Figure 1 As shown, the following steps are included: Prepare the raw materials according to the ratio; Thyme essential oil, modified resveratrol, and modified epigallocatechin gallate were mixed and subjected to ultrasonic emulsification (power 500 W, time 30 min); Add modified chitosan, sodium alginate-zinc ion gel microspheres, and mussel mucin, and stir to mix at 6°C; The mixture was treated with a pulsed electric field (field strength 8 kV / cm, time 20 ms).

[0038] Wherein, the preparation method of the modified resveratrol is as follows: Resveratrol and quercetin were added to a ball mill at a molar ratio of 1:1.3, and 15 wt% anhydrous calcium sulfate was added; Under nitrogen protection, ball milling was performed at 600 rpm for 5 h, and the temperature was controlled at ≤40°C; The eutectic powder was placed in an argon / oxygen (9:1) low-temperature plasma for 4 min (power 60 W); The template was eluted with supercritical CO2 (pressure 30 MPa, temperature 45°C).

[0039] Furthermore, the preparation method of the modified chitosan is as follows: Chitosan with a deacetylation degree of ≥95% was dissolved in 1.5% formic acid solution to prepare a 4 wt% solution; Sodium tripolyphosphate (the molar ratio of chitosan: sodium tripolyphosphate is 1:0.3) was added and stirred at 55°C for 50 min to activate the amino groups.

[0040] Furthermore, the preparation method of the sodium alginate-zinc ion gel microspheres is as follows: Internal phase: sodium alginate solution (4%) and zinc oxide nanorods (aspect ratio 25:1, 7%); External phase: soybean oil containing 3% sorbitan oleate and 0.7% ferroferric oxide nanoparticles (12 nm); dripping through a coaxial microfluidic chip (inner phase flow rate 0.6 mL / h, outer phase flow rate 6 mL / h); The droplets are oriented in a rotating magnetic field (2500 Gs, 120 rpm) to form a vertical channel structure, which increases the drug loading capacity; Cross-link and cure in 3% calcium chloride solution for 40 minutes.

[0041] In this embodiment, the preparation method of the modified epigallocatechin gallate is as follows: Epigallocatechin gallate, L-arginine, and zinc chloride were reacted in a pH 7.6 buffer solution at a molar ratio of 1:1.1:0.6; Salmonella lipopolysaccharide (template molecule, 1.5% by mass) was added, and ethyl orthosilicate was added dropwise for sol-gel embedding; The template was eluted with a 0.15 M sodium citrate / ethylenediaminetetraacetic acid mixture; Modified thiol polyethylene glycol (SHPEG, molecular weight 5000Da, grafting amount 20%).

[0042] Example 5 The embodiment of the present invention provides a preparation for laying hen salpingitis, comprising the following raw materials in parts by weight: 8 parts of thyme essential oil, 6 parts of modified resveratrol, 12 parts of modified chitosan, 15 parts of sodium alginate zinc ion gel microspheres, 7 parts of modified epigallocatechin gallate, and 10 parts of mussel mucin. The preparation method of the preparation for laying hen salpingitis is as follows: Figure 1 As shown, the following steps are included: Prepare the raw materials according to the ratio; Thyme essential oil, modified resveratrol, and modified epigallocatechin gallate were mixed and subjected to ultrasonic emulsification (power 500 W, time 30 min); Add modified chitosan, sodium alginate-zinc ion gel microspheres, and mussel mucin, and stir to mix at 6°C; The mixture was treated with a pulsed electric field (field strength 8 kV / cm, time 20 ms).

[0043] Wherein, the preparation method of the modified resveratrol is as follows: Resveratrol and quercetin were added to a ball mill at a molar ratio of 1:1.3, and 15 wt% anhydrous calcium sulfate was added; Under nitrogen protection, ball milling was performed at 600 rpm for 5 h, and the temperature was controlled at ≤40°C; The eutectic powder was placed in an argon / oxygen (9:1) low-temperature plasma for 4 min (power 60 W); The template was eluted with supercritical CO2 (pressure 30 MPa, temperature 45°C).

[0044] Furthermore, the preparation method of the modified chitosan is as follows: Chitosan with a deacetylation degree of ≥95% was dissolved in 1.5% formic acid solution to prepare a 4 wt% solution; Sodium tripolyphosphate (the molar ratio of chitosan: sodium tripolyphosphate is 1:0.3) was added and stirred at 55°C for 50 min to activate the amino groups.

[0045] Furthermore, the preparation method of the sodium alginate-zinc ion gel microspheres is as follows: Internal phase: sodium alginate solution (4%) and zinc oxide nanorods (aspect ratio 25:1, 7%); External phase: soybean oil containing 3% sorbitan oleate and 0.7% ferroferric oxide nanoparticles (12 nm); dripping through a coaxial microfluidic chip (inner phase flow rate 0.6 mL / h, outer phase flow rate 6 mL / h); The droplets are oriented in a rotating magnetic field (2500 Gs, 120 rpm) to form a vertical channel structure, which increases the drug loading capacity; Cross-link and cure in 3% calcium chloride solution for 40 minutes.

[0046] In this embodiment, the preparation method of the modified epigallocatechin gallate is as follows: Epigallocatechin gallate, L-arginine, and zinc chloride were reacted in a pH 7.6 buffer solution at a molar ratio of 1:1.1:0.6; Salmonella lipopolysaccharide (template molecule, 1.5% by mass) was added, and ethyl orthosilicate was added dropwise for sol-gel embedding; The template was eluted with a 0.15 M sodium citrate / ethylenediaminetetraacetic acid mixture; Modified thiol polyethylene glycol (SHPEG, molecular weight 5000Da, grafting amount 20%).

[0047] Example 6 (the difference from Example 3 is that a freeze-thaw cycle step is added) The embodiment of the present invention provides a preparation for laying hen salpingitis, comprising the following raw materials in parts by weight: 6.5 parts of thyme essential oil, 4.5 parts of modified resveratrol, 10 parts of modified chitosan, 12.5 parts of sodium alginate-zinc ion gel microspheres, 5.5 parts of modified epigallocatechin gallate, and 7.5 parts of mussel mucin. The preparation method of the preparation for laying hen salpingitis is as follows: Figure 2 As shown, the following steps are included: Prepare the raw materials according to the ratio; Thyme essential oil, modified resveratrol, and modified epigallocatechin gallate were mixed and subjected to ultrasonic emulsification (power 400 W, time 20 min); Add modified chitosan, sodium alginate-zinc ion gel microspheres, and mussel mucin, and stir to mix at 5°C; The mixture was treated with a pulsed electric field (field strength 6.5 kV / cm, time 15 ms); The mixture after pulse electric field treatment was frozen at −50°C for 2 h and then thawed at 25°C for 1 h, and the cycle was repeated twice.

[0048] Example 7 (the difference from Example 3 is that a freeze-thaw cycle step is added) The embodiment of the present invention provides a preparation for laying hen salpingitis, comprising the following raw materials in parts by weight: 6.5 parts of thyme essential oil, 4.5 parts of modified resveratrol, 10 parts of modified chitosan, 12.5 parts of sodium alginate-zinc ion gel microspheres, 5.5 parts of modified epigallocatechin gallate, and 7.5 parts of mussel mucin. The preparation method of the preparation for laying hen salpingitis is as follows: Figure 2 As shown, the following steps are included: Prepare the raw materials according to the ratio; Thyme essential oil, modified resveratrol, and modified epigallocatechin gallate were mixed and subjected to ultrasonic emulsification (power 400 W, time 20 min); Add modified chitosan, sodium alginate-zinc ion gel microspheres, and mussel mucin, and stir to mix at 5°C; The mixture was treated with a pulsed electric field (field strength 6.5 kV / cm, time 15 ms); The mixture after pulse electric field treatment was frozen at -45°C for 3 h and then thawed at 28°C for 1 h, and the cycle was repeated three times.

[0049] Example 8 (the difference from Example 3 is that a freeze-thaw cycle step is added) The embodiment of the present invention provides a preparation for laying hen salpingitis, comprising the following raw materials in parts by weight: 6.5 parts of thyme essential oil, 4.5 parts of modified resveratrol, 10 parts of modified chitosan, 12.5 parts of sodium alginate-zinc ion gel microspheres, 5.5 parts of modified epigallocatechin gallate, and 7.5 parts of mussel mucin. The preparation method of the preparation for laying hen salpingitis is as follows: Figure 2 As shown, the following steps are included: Prepare the raw materials according to the ratio; Thyme essential oil, modified resveratrol, and modified epigallocatechin gallate were mixed and subjected to ultrasonic emulsification (power 400 W, time 20 min); Add modified chitosan, sodium alginate-zinc ion gel microspheres, and mussel mucin, and stir to mix at 5°C; The mixture was treated with a pulsed electric field (field strength 6.5 kV / cm, time 15 ms); The mixture after pulse electric field treatment was frozen at -40°C for 4 h and then thawed at 30°C for 1 h, and the cycle was repeated 4 times.

[0050] Comparative Example 1: Commercially available antibiotic group (10 mg / kg enrofloxacin) Comparative Example 2: The difference from Example 7 is that the modified resveratrol is replaced by ordinary resveratrol.

[0051] Comparative Example 3: The difference from Example 7 is that the modified chitosan is replaced by ordinary chitosan.

[0052] Comparative Example 4: The difference from Example 7 is that the modified epigallocatechin gallate is replaced by ordinary epigallocatechin gallate.

[0053] Comparative Example 5: The difference from Example 7 is that the sodium alginate-zinc ion gel microspheres are replaced by ordinary calcium alginate microspheres.

[0054] Comparative Example 6: The difference from Example 7 is that no mussel mucin is contained.

[0055] Control group A: untreated group with infected salpingitis.

[0056] Control group B: healthy uninfected group.

[0057] Experimental testing 1. Animal Model 900 25-week-old Hy-Line Brown laying hens (salpingitis induced by Salmonella challenge) were randomly divided into 16 groups, 50 birds in each group; the dosage regimen was 0.5g preparation / feather / day, mixed with feed and fed for 7 days. 2. Test indicators Clinical efficacy: cure rate (disappearance of oviduct mucosal congestion / secretion), egg production recovery rate Mechanism verification: tissue drug concentration (HPLC), bacterial load (qPCR), inflammatory factors (TNF-α / IL-1β ELISA) Pharmacokinetics: Fallopian tube targeting coefficient (AUC in fallopian tube tissue 0-24h / Plasma AUC 0-24h ) The test data is shown in Table 1 below: Table 1 Performance test results Group Cure rate (%) Egg production recovery rate (%) Drug concentration in fallopian tube (μg / g) Bacterial load (lg CFU / g) TNF-α (pg / mg) Targeting coefficient Microsphere porosity (%) Example 1 75.3±4.0 77.6±3.4 14.2±1.5 3.0±0.3 58.7±5.8 4.3 38.2±2.9 Example 2 80.1±3.6 79.8±3.1 16.0±1.7 2.7±0.3 49.2±4.7 4.8 40.5±3.0 Example 3 87.5±3.2 83.4±2.9 18.3±2.1 2.3±0.2 39.8±4.3 5.1 42.3±3.1 Example 4 84.6±3.4 81.7±3.0 19.5±2.2 2.4±0.2 42.6±4.5 5.0 45.1±3.3 Example 5 78.9±3.8 78.3±3.3 15.8±1.8 2.8±0.3 53.4±5.2 4.5 39.7±2.8 Example 6 89.2±2.9 84.3±2.7 22.1±2.4 2.0±0.2 35.4±3.8 6.1 61.5±4.2 Example 7 92.5±2.7 86.1±2.3 25.7±2.8 1.8±0.2 31.6±3.6 6.9 68.7±4.5 Example 8 90.7±2.8 85.2±2.5 23.6±2.6 1.9±0.2 33.1±3.7 5.7 72.3±4.8 Comparative Example 1 78.4±4.2 76.5±3.8 9.2±1.1 3.1±0.3 65.4±6.7 0.8 - Comparative Example 2 53.7±4.3 68.9±3.5 8.6±1.0 4.7±0.4 89.2±8.1 2.1 65.2±4.0 Comparative Example 3 64.2±4.1 73.6±3.3 11.4±1.3 3.9±0.3 72.5±6.9 2.8 66.8±4.3 Comparative Example 4 76.8±3.7 79.1±3.0 15.3±1.7 3.2±0.3 57.3±5.6 4.1 67.5±4.2 Comparative Example 5 61.5±4.5 71.2±3.6 7.9±0.9 4.3±0.4 81.7±7.8 1.5 41.8±3.6 Comparative Example 6 80.3±3.5 81.7±2.8 14.6±1.6 2.7±0.3 48.5±4.9 3.2 66.1±4.1 Control group A 0 52.3±3.1 - 7.8±0.4 128.6±12.3 - - Control group B - 86.7±2.5 - ND 28.3±3.2 - - Note: ND means no bacteria detected From the above results, it can be seen that the preparation prepared by the present invention has a good effect on preventing and treating salpingitis.

[0058] It should be noted that, for the sake of simplicity, the aforementioned embodiments are described as a series of action combinations. However, those skilled in the art should be aware that the present invention is not limited by the order of the actions described, because according to the present invention, certain steps may be performed in other orders or simultaneously. Secondly, those skilled in the art should also be aware that the embodiments described in this specification are all preferred embodiments, and the actions and modules involved are not necessarily required by the present invention.

[0059] In the embodiments provided in this application, it should be understood that the disclosed devices can be implemented in other ways. For example, the device embodiments described above are merely schematic. For example, the division of the above-mentioned units is only a logical function division. In actual implementation, there may be other division methods, such as multiple units or components can be combined or integrated into another system, or some features can be ignored or not executed. Another point is that the mutual coupling or communication connection shown or discussed can be through some interfaces, and the indirect coupling or communication connection between devices or units can be in the form of telecommunications or other forms.

[0060] The units described above as separate components may or may not be physically separate, and the components shown as units may or may not be physical units, that is, they may be located in one place or distributed across multiple network units. Some or all of these units may be selected according to actual needs to achieve the purpose of the solution of this embodiment.

[0061] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the scope of protection of the invention. Obviously, the embodiments described are only some embodiments of the present invention, rather than all embodiments. Based on these embodiments, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention. Although the present invention has been described in detail with reference to the above embodiments, ordinary technicians in this field can still combine, add, delete or make other adjustments to the features in the various embodiments of the present invention according to the circumstances without conflict, without making creative work, so as to obtain different other technical solutions that do not deviate from the concept of the present invention in essence, and these technical solutions also fall within the scope of protection of the present invention.

Claims

1. A preparation for laying hen salpingitis, characterized in that: The invention comprises the following raw materials in parts by weight: 5-8 parts of thyme essential oil, 3-6 parts of modified resveratrol, 8-12 parts of modified chitosan, 10-15 parts of sodium alginate-zinc ion gel microspheres, 4-7 parts of modified epigallocatechin gallate, and 5-10 parts of mussel mucin.

2. The preparation for laying hen salpingitis according to claim 1, wherein The invention comprises the following raw materials in parts by weight: 6-7 parts of thyme essential oil, 4-5 parts of modified resveratrol, 9-11 parts of modified chitosan, 11-14 parts of sodium alginate-zinc ion gel microspheres, 5-6 parts of modified epigallocatechin gallate, and 6-9 parts of mussel mucin.

3. The preparation for laying hen salpingitis according to claim 2, wherein The invention comprises the following raw materials in parts by weight: 6.5 parts of thyme essential oil, 4.5 parts of modified resveratrol, 10 parts of modified chitosan, 12.5 parts of sodium alginate-zinc ion gel microspheres, 5.5 parts of modified epigallocatechin gallate, and 7.5 parts of mussel mucin.

4. The preparation for laying hen salpingitis according to claim 1, wherein The preparation method of the modified resveratrol is as follows: Resveratrol and quercetin were added to a ball mill at a molar ratio of 1:1.1-1.3, and 5-15 wt% anhydrous calcium sulfate was added; Under nitrogen protection, ball mill at 400-600 rpm for 3-5 hours, controlling the temperature ≤40°C; The eutectic powder was placed in an argon / oxygen low-temperature plasma for 2-4 minutes; The template is eluted with supercritical CO2.

5. The preparation for laying hen salpingitis according to claim 1, characterized in that The preparation method of the modified chitosan is as follows: Dissolve chitosan with a deacetylation degree of ≥95% in 0.5-1.5% formic acid solution to prepare a 2-4wt% solution; Add sodium tripolyphosphate and stir at 45-55°C for 30-50 minutes to activate the amino group.

6. The preparation for laying hen salpingitis according to claim 5, characterized in that The molar ratio of chitosan to sodium tripolyphosphate is 1:0.1-0.

3.

7. The preparation for laying hen salpingitis according to claim 1, characterized in that The preparation method of the sodium alginate-zinc ion gel microspheres is as follows: Internal phase: sodium alginate solution and zinc oxide nanorods; External phase: soybean oil containing 1-3% sorbitan oleate and 0.3-0.7% ferroferric oxide nanoparticles; dripping through a coaxial microfluidic chip; The droplets are oriented and aligned by a rotating magnetic field; Cross-link and cure in 1-3% calcium chloride solution for 20-40 minutes.

8. The preparation for laying hen salpingitis according to claim 7, characterized in that The preparation method of the modified epigallocatechin gallate is as follows: Epigallocatechin gallate, L-arginine, and zinc chloride are reacted in a pH 7.2-7.6 buffer solution at a molar ratio of 1:0.9-1.1:0.4-0.6; Salmonella lipopolysaccharide was added, and ethyl orthosilicate was added dropwise for sol-gel embedding; The template was eluted with a 0.05-0.15 M sodium citrate / ethylenediaminetetraacetic acid mixture; Modified with thiol polyethylene glycol.

9. The method for preparing the preparation for laying hen salpingitis according to any one of claims 1 to 8, wherein: The steps include: Prepare the raw materials according to the ratio; Thyme essential oil, modified resveratrol, and modified epigallocatechin gallate were mixed and subjected to ultrasonic emulsification; Add modified chitosan, sodium alginate-zinc ion gel microspheres, and mussel mucin, and stir to mix at 4-6°C; The mixture is treated with a pulsed electric field.

10. The method for preparing the preparation for laying hen salpingitis according to claim 9, wherein: After the mixture is treated with a pulsed electric field, it is frozen at -50 to -40°C for 2 to 4 hours, then thawed at 25 to 30°C for 1 hour, and the cycle is repeated 2 to 4 times to form a porous network structure.

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