Traditional Chinese medicine composition for treating porcine reproductive and respiratory syndrome and preparation method thereof

Abstract

The invention belongs to the technical field of medicine preparation, and particularly relates to a traditional Chinese medicine composition for treating porcine reproductive and respiratory syndrome and a preparation method thereof. The preparation method of the traditional Chinese medicine composition comprises the following preparation steps: mixing and crushing traditional Chinese medicine materials, adding a fermentation strain into the crushed traditional Chinese medicine materials, and fermenting for 3-5 days to obtain a traditional Chinese medicine fermentation mixture; adding a compound enzyme into the yeast powder, adjusting the pH value to 6-9, carrying out enzymolysis at 40-60 DEG C for 3-5 hours, and carrying out enzyme deactivation and drying to obtain a yeast peptide solution; uniformly mixing an ascorbic acid solution with a sodium selenite solution, adjusting the pH value of the mixed system, adding the yeast peptide liquid into the mixed system, uniformly mixing, adding gelatin, uniformly mixing, and drying to obtain a yeast peptide compound; and uniformly mixing the traditional Chinese medicine fermentation mixture with the yeast peptide compound to prepare the traditional Chinese medicine composition for treating the porcine reproductive and respiratory syndrome. The invention provides a traditional Chinese medicine and polypeptide formula which has a better curative effect on the porcine reproductive and respiratory syndrome.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical preparation, and particularly relates to a traditional Chinese medicine composition for treating porcine reproductive and respiratory syndrome (PRRS) and a preparation method thereof. Background Art

[0002] PRRS is the number one killer that destroys the immune system of pigs. The problem of PRRS has been plaguing farmers and has brought huge losses to many pig farms. Porcine reproductive and respiratory syndrome (PRRS), also known as swine reproductive and respiratory syndrome, has an adverse impact on sows, piglets, weaned piglets and adult fattening pigs. It is caused by the porcine reproductive and respiratory syndrome virus in the family Arteriviridae, and is mainly characterized by reproductive disorders, dyspnea, blue-purple ears, immune decline, and the occurrence of other diseases. At present, the main method for preventing PRRS is vaccination, but an ideal PRRS vaccine has not been developed yet, and the protective ability of the vaccine and the prevention of breakthrough infections still need to be improved. In addition, there is no antiviral drug directly targeting PRRS on the market at present. There are only reports on purifying PRRS in pig farms through traditional Chinese medicine and antibiotics such as tilmicosin. Although the use of antibiotics has a certain effect in treatment, the disease will recur soon after stopping the drug.

[0003] Patent CN 118903077 A discloses the application of shikonin and its analogs in the preparation of drugs against porcine reproductive and respiratory syndrome virus. The shikonin analogs are deoxyshikonin, acetylshikonin or β,β-dimethylacryloylshikonin, and an anti-porcine reproductive and respiratory syndrome virus drug and a composition against PRRSV are disclosed, which can be used for preventing PRRSV infection and / or treating anti-porcine reproductive and respiratory syndrome caused by PRRSV. Although this invention tested the cell viability of Marc-145 cells at different shikonin concentrations, the inhibition rate of PRRSV at different shikonin concentrations, and the antiviral test results of animal sera, it lacked actual clinical experiments on pigs and was still a long way from being marketed. In addition, when using some traditional Chinese medicines to treat PRRS, traditional Chinese medicines are mostly directly added after being pulverized, which has disadvantages such as poor palatability, large addition amount, slow onset, etc. in actual application, and the treatment effect is not ideal.

[0004] Therefore, it is of great significance to the breeding industry to develop a drug that can effectively treat PRRS. Summary of the Invention

[0005] In view of the deficiencies in the prior art, the present invention provides a traditional Chinese medicine composition for treating PRRS and a preparation method thereof. The present invention discloses a formula of traditional Chinese medicine materials, and compounded with polypeptide substances to optimize the formula. The prepared traditional Chinese medicine composition has a good curative effect on PRRS.

[0006] To achieve the above object, the technical scheme adopted by the present invention is as follows:

[0007] The first aspect of the present invention provides a method for preparing a Chinese medicine composition for treating blue ear disease in pigs, comprising the following preparation steps:

[0008] (1) mixing and crushing Chinese medicinal materials, adding fermentation bacteria thereto, and fermenting for 3-5 days to obtain a Chinese medicinal fermentation mixture;

[0009] (2) adding complex enzyme to yeast powder, adjusting the pH to 6-9, performing enzymolysis at 40-60° C. for 3-5 hours, inactivating the enzyme, and drying to obtain yeast peptide liquid;

[0010] (3) taking an ascorbic acid solution and a sodium selenite solution and mixing them evenly, adjusting the pH of the mixed system to 7.5-8.5, adding the yeast peptide solution thereto, mixing evenly, then adding gelatin and mixing evenly, and drying to obtain a yeast peptide complex;

[0011] (4) Evenly mixing the Chinese medicine fermentation mixture of step (1) and the yeast peptide complex of step (4) to prepare the Chinese medicine composition for treating porcine blue ear disease.

[0012] In some embodiments, the Chinese medicinal materials in step (1) include, by weight, 10-20 parts of Polygonum multiflorum, 5-15 parts of Paeonia suffruticosa, 10-30 parts of Astragalus, 10-25 parts of Pulsatilla scabra, 1-10 parts of Sophora flavescens, 1-15 parts of Ephedra root, 20-30 parts of Coix seeds, 10-30 parts of Licorice, 5-20 parts of Isatis indigotica, 1-10 parts of Notopterygium wilfordii, 10-20 parts of Chuanxiong, 5-15 parts of Cinnamomum cassia, 10-25 parts of White Peony Root, 5-20 parts of Drynaria ginger, 10-20 parts of Saposhnikovia divaricata, 5-25 parts of Cyperus rotundus, 10-20 parts of Stir-fried Atractylodes macrocephala, and 1-10 parts of Codonopsis pilosula.

[0013] In some embodiments, the fermentation bacteria in step (1) is Bacillus coagulans.

[0014] In some embodiments, the composite enzyme of step (2) comprises papain and trypsin in a mass ratio of (2.5-3.5):1.

[0015] In some embodiments, the amount of the complex enzyme is 0.2-0.5 wt % of the yeast powder.

[0016] In some embodiments, the molar ratio of ascorbic acid to sodium selenite in step (3) is (4-6):1.

[0017] In some embodiments, the amount of gelatin used in step (3) is 5-10 wt % of the yeast peptide solution.

[0018] In some embodiments, the mass ratio of the traditional Chinese medicine fermentation mixture and the yeast peptide complex in step (4) is (2-3):1.

[0019] In the second aspect of the present invention, there is provided a traditional Chinese medicine composition for treating porcine reproductive and respiratory syndrome (PRRS) prepared by the preparation method described in the above scheme.

[0020] In some embodiments, the dosage of the traditional Chinese medicine composition is 0.1-0.5 wt% of the feed used.

[0021] Through repeated experiments, the present invention obtains a formula of traditional Chinese medicine materials, which has a good curative effect on porcine reproductive and respiratory syndrome. The traditional Chinese medicine formula contains 18 kinds of medicinal materials, namely bistort, cortex moutan, astragalus membranaceus, pulsatilla chinensis, sophora flavescens, ephedra root, coix seed, liquorice root, folium isatidis, notopterygium root, chuanxiong rhizome, cassia twig, white peony root, dried ginger, ledebouriella root, dryopteris crassirhizoma, stir-fried atractylodes macrocephala, and codonopsis pilosula. The functions of each medicinal material are as follows: Bistort: clearing heat and detoxifying, cooling blood for hemostasis, suppressing liver wind, and dissipating carbuncles and nodules; Cortex moutan: clearing heat and cooling blood, promoting blood circulation to remove stasis, and removing deficiency heat; Astragalus membranaceus: replenishing qi and ascending yang, strengthening the exterior and consolidating the superficies, promoting diuresis and alleviating edema; Pulsatilla chinensis: clearing heat and detoxifying, cooling blood, improving eyesight, and drying dampness; Sophora flavescens: clearing heat and drying dampness, treating damp-heat dysentery, hematochezia, and jaundice; Ephedra root: consolidating the superficies to stop sweating, promoting diuresis and alleviating edema, relieving cough and asthma; Coix seed: promoting diuresis and percolating dampness, strengthening the spleen and stopping diarrhea, treating diarrhea due to spleen deficiency; Liquorice root: invigorating the spleen and replenishing qi, resolving phlegm and relieving cough, and coordinating the properties of various medicinal materials; Folium isatidis: clearing heat and detoxifying, cooling blood and eliminating macules, and dispelling pathogenic factors on the exterior; Notopterygium root: dispersing wind-cold, expelling wind-dampness, and relieving pain; Chuanxiong rhizome: promoting blood circulation and qi movement, dispelling wind and relieving pain, being the qi drug in blood; Cassia twig: inducing sweating and releasing the exterior, warming the channels and promoting qi movement, and assisting yang in transforming qi; White peony root: tonifying the kidney and strengthening the spleen, nourishing blood and regulating menstruation, and suppressing liver yang; Dried ginger: warming the middle-jiao to dispel cold, restoring yang and unblocking meridians, and warming the lungs to resolve fluid-retention; Ledebouriella root: pungent and sweet in flavor, dispersing wind and relieving the exterior, expelling wind-dampness; Dryopteris crassirhizoma: cooling blood for hemostasis, clearing heat and detoxifying; Stir-fried Atractylodes macrocephala: strengthening the spleen and replenishing qi, drying dampness and promoting diuresis, replenishing qi of the spleen and stopping diarrhea; Codonopsis pilosula: replenishing middle-jiao qi, promoting the production of body fluid and nourishing blood, and moistening the lungs to promote the production of body fluid.

[0022] In order to further improve the medicinal effect, the present invention selects Bacillus coagulans to ferment the traditional Chinese medicinal materials. On the one hand, Bacillus coagulans can produce various enzymes during the fermentation process, such as cellulase, amylase, protease, etc. These enzymes can decompose components such as cellulose, starch, and protein in the mixed medicinal materials. For medicinal materials rich in polysaccharides and proteins such as Astragalus membranaceus and Codonopsis pilosula, the enzymatic hydrolysis can make the active ingredients such as Astragalus polysaccharides and Codonopsis pilosula saponins dissolve better, improving the utilization rate of the medicinal materials; on the other hand, the fermentation of Bacillus coagulans can improve the taste and palatability of the medicinal materials. The fermented medicinal materials may produce some special flavor substances, improving the originally bitter taste of the mixed medicinal materials. For pigs, it can improve their acceptance of the medicine, ensuring that pigs can ingest enough medicine to treat porcine reproductive and respiratory syndrome. In addition, organic acids such as lactic acid and butyric acid are produced during the fermentation of Bacillus coagulans. These organic acids can adjust the pH value of the fermentation system, creating an acidic environment conducive to the stable existence of drug components; at the same time, the organic acids also have certain antibacterial activities, which can inhibit the growth of harmful bacteria, providing protection for the intestinal health of pigs. Moreover, these metabolites may also interact with the components in the medicinal materials to produce new substances with pharmacological activities. The toxic substances in the fermented traditional Chinese medicinal materials are relatively reduced, which can regulate the intestinal microecology of pigs, enhance the immune function of pigs, and improve the physiological state of pigs, increasing the disease resistance of pigs.

[0023] Since the illness may lead to a decrease in the absorption and utilization rate of traditional Chinese medicine by the pig's gastrointestinal tract, to address this issue, the present invention uses a compound enzyme (papain and trypsin) to enzymatically hydrolyze yeast powder. Papain mainly acts on the carboxyl-side peptide bonds of basic amino acid residues such as arginine and lysine, and trypsin specifically hydrolyzes the carboxyl-side peptide bonds of lysine and arginine. During the enzymatic hydrolysis process, these enzymes cut the peptide bonds in yeast protein, gradually decomposing the large-molecule yeast protein into smaller peptide segments. By adjusting the pH and temperature to promote the enzymatic hydrolysis reaction, and by controlling the enzymatic hydrolysis time, the molecular weight distribution of the polypeptide is further controlled, so that the obtained yeast peptide has good biological activity and absorbability. In order to improve the stability of yeast peptide, the present invention modifies the enzymatically hydrolyzed yeast peptide with ascorbic acid and sodium selenite under certain pH and raw material ratios. Among them, ascorbic acid can prevent the oxidation of selenium ions, promote the interaction between selenium ions and amino acid residues in yeast peptide, forming a complex or chelate, enhancing the stability and biological activity of yeast peptide. After adding gelatin, functional groups such as amino and carboxyl groups in the gelatin molecule will undergo physical adsorption or chemical cross-linking reactions with yeast peptide, forming a complex structure, and a stable yeast peptide complex is obtained after drying.

[0024] The yeast peptide complex and the traditional Chinese medicine fermentation mixture are compounded in proportion. Components such as astragalus polysaccharide and codonopsis pilosula saponin in the traditional Chinese medicine fermentation mixture can activate the immune system of pigs, promote the proliferation and differentiation of immune cells. The yeast peptides in the yeast peptide complex can provide nutritional support for immune cells and enhance the activity of immune cells. The synergistic effect of the two can more effectively regulate the immune system of pigs and enhance the resistance of pigs to porcine reproductive and respiratory syndrome virus. In addition, the yeast peptides in the yeast peptide complex can promote the absorption of the effective components of traditional Chinese medicine by the intestine. For example, yeast peptides can regulate the function of intestinal mucosal cells, increase the permeability of cell membranes, so that the effective components such as polysaccharides, saponins, and amino acids in the traditional Chinese medicine fermentation mixture can better enter the blood circulation and play a therapeutic role. At the same time, components such as gelatin in the yeast peptide complex can also slowly release yeast peptides and adsorbed traditional Chinese medicine components in the gastrointestinal tract, extend their action time in the intestine, improve the absorption efficiency, and maintain the balance of the intestinal microecology.

[0025] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0026] 1. The present invention obtains a traditional Chinese medicine formula through repeated experiments, adds polypeptide substances thereto, optimizes the formula, and improves the treatment effect. The obtained traditional Chinese medicine composition has a good curative effect on porcine reproductive and respiratory syndrome.

[0027] 2. The present invention selects Bacillus coagulans to ferment the traditional Chinese medicine materials, so that the effective components therein can be better dissolved, the utilization rate of the medicinal materials is improved, and the originally possibly bitter taste of the mixed medicinal materials is improved.

[0028] 3. The present invention uses a composite enzyme (papain and trypsin) to enzymatically hydrolyze yeast powder to obtain yeast peptides with good biological activity and absorbability, and uses ascorbic acid and sodium selenite to modify the enzymatically hydrolyzed yeast peptides to enhance the stability and biological activity of the yeast peptides and improve the drug efficacy. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 For the pathogen molecular detection results of pigs in Clinical Experiment 1;

[0030] Figure 2 For the homology comparison results of the field strain and the common strain in Clinical Experiment 1;

[0031] Figure 3 For the porcine reproductive and respiratory syndrome antibody detection results of pigs before treatment in Clinical Experiment 2;

[0032] Figure 4 For the porcine reproductive and respiratory syndrome antibody detection results of pigs after treatment in Clinical Experiment 2. DETAILED DESCRIPTION OF THE INVENTION

[0033] The various exemplary embodiments of the present invention will be described in detail below. This detailed description should not be construed as a limitation on the present invention, but rather as a more detailed description of certain aspects, features, and implementation schemes of the present invention. It should be understood that the terms described in the present invention are only for describing specific embodiments and are not used to limit the present invention. Additionally, for the numerical ranges in the present invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Each intermediate value within any stated value or stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0034] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention pertains. Although the present invention only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. Without departing from the scope or spirit of the present invention, various improvements and changes can be made to the specific embodiments of the present invention specification, which are obvious to those skilled in the art. Other embodiments obtained from the specification of the present invention are obvious to those skilled in the art. The specification and examples of the present application are merely exemplary.

[0035] Regarding the use of "comprising", "including", "having", "containing", etc. herein, they are all open-ended terms, meaning including but not limited to.

[0036] It should be noted that operations such as "grinding", "centrifuging", "stirring", etc. described in the present invention are conventional operations for those skilled in the art and can be selected according to actual operations.

[0037] The parts by mass described in the present invention can be parts by grams, kilograms, tons, or other mass units.

[0038] The preservation number of Bacillus coagulans used in the present invention is CICC 24625, and the activity of Bacillus coagulans is 10 billion cfu / g; the enzyme activities of papain and trypsin are both 100,000 U / g.

[0039] Example 1

[0040] A preparation method of a traditional Chinese medicine composition for treating porcine reproductive and respiratory syndrome, comprising the following preparation steps:

[0041] (1) Mix and pulverize traditional Chinese medicine materials and pass through a 60-mesh sieve, add 1 wt% of Bacillus coagulans thereto, and ferment at 37 °C and 60% humidity for 5 days to obtain a traditional Chinese medicine fermentation mixture;

[0042] (2) adding 0.2 wt% of complex enzyme and 3 times the weight of pure water to yeast powder, adjusting the pH to 6, performing enzymolysis at 40° C. for 5 h, and inactivating the enzyme at 100° C. for 10 min to obtain yeast peptide liquid;

[0043] (3) 100 mL of a 2 mol / L ascorbic acid aqueous solution and 100 mL of a 0.5 mol / L sodium selenite aqueous solution were mixed evenly, the pH of the mixed system was adjusted to 7.5, 70 g of the yeast peptide solution stirred evenly in step (1) was added thereto, mixed evenly, 7 g of gelatin was added, mixed evenly, and spray-dried to obtain a yeast peptide complex;

[0044] (4) 20 g of the traditional Chinese medicine fermentation mixture of step (1) and 10 g of the yeast peptide complex of step (4) are mixed evenly to prepare the traditional Chinese medicine composition for treating porcine blue ear disease.

[0045] The Chinese medicinal materials used, measured by weight, include 10 parts of polygonum multiflorum, 5 parts of peony bark, 10 parts of astragalus, 10 parts of pulsatilla, 1 part of sophora flavescens, 1 part of ephedra root, 20 parts of coix seed, 10 parts of liquorice, 5 parts of isatis indigotica, 1 part of notopterygium wilfordii, 10 parts of chuanxiong, 5 parts of cassia twig, 10 parts of white peony root, 5 parts of dried ginger, 10 parts of siler, 5 parts of cyperus rotundus, 10 parts of stir-fried atractylodes, and 1 part of codonopsis pilosula.

[0046] The complex enzyme used contained papain and trypsin in a mass ratio of 2.5:1.

[0047] Example 2

[0048] A method for preparing a traditional Chinese medicine composition for treating blue ear disease in pigs, comprising the following preparation steps:

[0049] (1) mixing Chinese medicinal materials, crushing them through a 100-mesh sieve, adding 3 wt % of Bacillus coagulans thereto, and fermenting them at 50° C. and 70% humidity for 3 days to obtain a Chinese medicinal fermentation mixture;

[0050] (2) adding 0.5 wt% of complex enzyme and 5 times the weight of pure water to yeast powder, adjusting the pH to 9, performing enzymolysis at 60° C. for 3 h, and inactivating the enzyme at 100° C. for 10 min to obtain yeast peptide liquid;

[0051] (3) 100 mL of a 2 mol / L ascorbic acid aqueous solution and 100 mL of a 0.35 mol / L sodium selenite aqueous solution were mixed uniformly, and after adjusting the pH of the mixed system to 8.5, 210 g of the yeast peptide solution stirred uniformly in step (1) was added thereto, mixed uniformly, and then 12 g of gelatin was added and mixed uniformly, and spray-dried to obtain a yeast peptide complex;

[0052] (4) Mix 30 g of the traditional Chinese medicine fermentation mixture from step (1) with 10 g of the yeast peptide complex from step (4) evenly to prepare the traditional Chinese medicine composition for treating porcine reproductive and respiratory syndrome.

[0053] The traditional Chinese medicine materials used, by mass parts, include 20 parts of bistort rhizome, 15 parts of moutan bark, 30 parts of astragalus root, 25 parts of pulsatilla root, 10 parts of lightyellow sophora root, 15 parts of ephedra root, 30 parts of coix seed, 30 parts of liquorice root, 20 parts of folium isatidis, 10 parts of incised notopterygium rhizome, 20 parts of chuanxiong rhizome, 15 parts of cassia twig, 25 parts of white peony root, 20 parts of dried ginger, 20 parts of divaricate saposhnikovia root, 25 parts of dryopteris crassirhizoma, 20 parts of stir-fried atractylodes macrocephala, and 10 parts of pilose asiabell root.

[0054] The composite enzyme used contains papain and trypsin with a mass ratio of 3:1.

[0055] Example 3

[0056] A preparation method of a traditional Chinese medicine composition for treating porcine reproductive and respiratory syndrome, comprising the following preparation steps:

[0057] (1) Mix the traditional Chinese medicine materials, crush them through an 80-mesh sieve, add 2 wt% of bacillus coagulans thereto, and ferment at 45°C and 65% humidity for 3 days to obtain a traditional Chinese medicine fermentation mixture;

[0058] (2) Add 0.35 wt% of the composite enzyme and 4 times the mass of pure water to yeast powder, adjust the pH to 8, enzymolyze at 50°C for 2 h, and inactivate the enzyme at 100°C for 10 min to obtain a yeast peptide solution;

[0059] (3) Take 100 mL of an aqueous solution of 2 mol / L ascorbic acid and 100 mL of an aqueous solution of 0.4 mol / L sodium selenite, mix them evenly, adjust the pH of the mixed system to 8, add 140 g of the yeast peptide solution stirred evenly from step (1) thereto, mix evenly, then add 10 g of gelatin and mix evenly, and perform spray drying to obtain a yeast peptide complex;

[0060] (4) Mix 25 g of the traditional Chinese medicine fermentation mixture from step (1) with 10 g of the yeast peptide complex from step (4) evenly to prepare the traditional Chinese medicine composition for treating porcine reproductive and respiratory syndrome.

[0061] The traditional Chinese medicine materials used, by mass parts, include 15 parts of bistort rhizome, 10 parts of moutan bark, 20 parts of astragalus root, 20 parts of pulsatilla root, 5 parts of lightyellow sophora root, 10 parts of ephedra root, 25 parts of coix seed, 20 parts of liquorice root, 12 parts of folium isatidis, 5 parts of incised notopterygium rhizome, 15 parts of chuanxiong rhizome, 10 parts of cassia twig, 20 parts of white peony root, 15 parts of dried ginger, 15 parts of divaricate saposhnikovia root, 15 parts of dryopteris crassirhizoma, 15 parts of stir-fried atractylodes macrocephala, and 5 parts of pilose asiabell root.

[0062] The composite enzyme used contains papain and trypsin with a mass ratio of 2.5:1.

[0063] Example 4

[0064] A method for preparing a traditional Chinese medicine composition for treating porcine blue ear disease, the specific implementation method is the same as that of Example 3, except that the concentration of the 100mL aqueous solution of sodium selenite used in step (3) is 1 mol / L.

[0065] Example 5

[0066] A method for preparing a traditional Chinese medicine composition for treating porcine blue ear disease, the specific implementation method is the same as that of Example 3, except that in step (2), the complex enzyme comprises papain and trypsin in a mass ratio of 4:1.

[0067] Comparative Example 1

[0068] A method for preparing a Chinese medicine composition for treating porcine blue ear disease, the specific implementation method is the same as that of Example 3, except that step (1) is: mixing Chinese medicinal materials, crushing them through an 80-mesh sieve, and obtaining a Chinese medicine mixture.

[0069] Comparative Example 2

[0070] A method for preparing a traditional Chinese medicine composition for treating blue ear disease in pigs, comprising the following preparation steps:

[0071] (1) mixing Chinese medicinal materials, crushing them through an 80-mesh sieve, adding 2 wt % of Bacillus coagulans thereto, and fermenting them at 45° C. and 65% humidity for 3 days to obtain a Chinese medicinal fermentation mixture;

[0072] (2) 100 mL of a 2 mol / L ascorbic acid aqueous solution and 100 mL of a 0.4 mol / L sodium selenite aqueous solution were mixed uniformly, and after adjusting the pH of the mixed system to 8, 140 g of a 20 wt% yeast powder solution dispersed in pure water was added thereto, mixed uniformly, and then 10 g of gelatin was added and mixed uniformly, and spray-dried to obtain a yeast peptide complex;

[0073] (3) 25 g of the traditional Chinese medicine fermentation mixture of step (1) and 10 g of the yeast peptide complex of step (4) are mixed evenly to prepare the traditional Chinese medicine composition for treating porcine blue ear disease.

[0074] The Chinese medicinal materials used, measured by weight, include 15 parts of polygonum multiflorum, 10 parts of peony bark, 20 parts of astragalus, 20 parts of pulsatilla, 5 parts of sophora flavescens, 10 parts of ephedra root, 25 parts of coix seed, 20 parts of liquorice, 12 parts of isatis indigotica, 5 parts of notopterygium, 15 parts of chuanxiong, 10 parts of cassia twig, 20 parts of white peony root, 15 parts of dried ginger, 15 parts of siler, 15 parts of cyperus rotundus, 15 parts of stir-fried atractylodes, and 5 parts of codonopsis pilosula.

[0075] The complex enzyme used contained papain and trypsin in a mass ratio of 2.5:1.

[0076] Comparative Example 3

[0077] A preparation method of a traditional Chinese medicine composition for treating porcine reproductive and respiratory syndrome (PRRS), the specific implementation manner is the same as that of Example 3, except that step (3) is: taking 100 mL of an aqueous solution of 2 mol / L ascorbic acid and 100 mL of an aqueous solution of 0.4 mol / L sodium selenite, mixing them evenly, adjusting the pH of the mixed system to 8, then adding 140 g of the evenly stirred yeast peptide solution in step (1) thereto, mixing evenly, and spray drying to obtain a yeast peptide complex.

[0078] Comparative Example 4

[0079] A preparation method of a traditional Chinese medicine composition for treating porcine reproductive and respiratory syndrome (PRRS), the specific implementation manner is the same as that of Example 3, except that papain of equal mass is used instead of the complex enzyme.

[0080] Comparative Example 5

[0081] A preparation method of a traditional Chinese medicine composition for treating porcine reproductive and respiratory syndrome (PRRS), the specific implementation manner is the same as that of Example 3, except that trypsin of equal mass is used instead of the complex enzyme.

[0082] Comparative Example 6

[0083] A preparation method of a traditional Chinese medicine composition for treating porcine reproductive and respiratory syndrome (PRRS) includes the following preparation steps:

[0084] Mix and pulverize traditional Chinese medicine materials through an 80-mesh sieve to obtain a traditional Chinese medicine composition.

[0085] Performance Test

[0086] 1. Antiviral Experiment

[0087] Respectively take the traditional Chinese medicine mixtures / traditional Chinese medicine compositions obtained in the examples and comparative examples with equal mass, add pure water covering each group of medicinal materials, heat and extract at 70 °C for 0.5 h, filter, repeat the extraction step 2 times, combine the extraction solutions, concentrate and dry them, and then prepare a standard solution with a concentration of 15 mg / mL with pure water.

[0088] Refer to Patent CN 118903077 A for the antiviral experiment: Seed Marc-145 cells at 1.5×10 4Cells / well (volume 100 μL) were inoculated into a 96-well cell culture plate. When the cells adhered and the confluence rate exceeded 90%, the medium was replaced with DMEM maintenance medium (DMEM medium, 2% serum, penicillin 100 U / mL, streptomycin 100 μg / mL). Each infection group was added with WUH3 porcine reproductive and respiratory syndrome virus at 0.1 MOI (multiplicity of infection). At the same time, each test solution was diluted into 6 concentration gradients with the maintenance medium and added to each drug treatment well (Examples 1-5, Comparative Examples 1-6). Each concentration gradient was set with 3 replicates, and their final concentrations were 10 mg / mL, 5 mg / mL, 2.5 mg / mL, 1 mg / mL, 0.1 mg / mL. They were cultured in a 37°C cell culture incubator for 60 h. The culture plate was taken out and frozen and thawed once at -20°C, and each well was pipetted and 180 μL was aspirated and added to the virus nucleic acid purification kit machine to automatically extract RNA.

[0089] For other operations, reference can be made to Patent CN 118903077 A

[0046] -

[0049] , and the specific test data are shown in Table 1.

[0090] Table 1

[0091]

[0092] As can be seen from Table 1, the traditional Chinese medicine mixtures prepared in Examples 1-3 have good inhibitory effects on porcine reproductive and respiratory syndrome virus, and their IC 50 is within 2.11-2.23 mg / mL; compared with Example 3, the preparation process of the yeast peptide complex used in Example 4 and Comparative Example 3 has changed, affecting its own stability and the bioavailability of traditional Chinese medicine components, resulting in a decrease in IC 50 ; in Comparative Example 2, yeast powder was used instead of the yeast peptide complex, and the preparation process of the yeast peptide complex used in Example 5 and Comparative Examples 4 and 5 has changed, which may change the molecular weight of the peptide and affect the efficacy of the drug; the traditional Chinese medicine materials used in Comparative Example 1 were not fermented, and Comparative Example 6 only contained crushed medicinal materials, and their drug effects were weaker than those of the traditional Chinese medicine compositions in Examples 1-3, and IC 50 increased significantly.

[0093] 2. Clinical trial

[0094] Clinical trial 1

[0095] Experimental site: In mid-April 2023, a large-scale farm in Bozhou, Anhui Province, with 530 sows in stock and a fattening farm (3000 in stock). The diseased pens had 600 pigs weighing about 30 kg;

[0096] According to feedback, sporadic deaths began to occur in mid-March. During that period, multiple treatment plans were used but the effects were not good, and the incidence and mortality rates showed an upward trend. The test report is shown inFigure 1 , 2 。

[0097] Medication plan: (1) Mixing in the whole flock: Add 3 kg of the traditional Chinese medicine composition prepared in Example 3, 1 kg of Anqile, and 1 kg of 20% tilmicosin to each ton of feed, and use continuously for 15 days; (2) Isolate the sick pigsty: Add medicine to the drinking water, add 100 g of the traditional Chinese medicine composition prepared in Example 3, 100 g of 20% tilmicosin, and 100 g of Anqile to every 100 catties of water; (3) Intramuscular injection for severely sick pigs: 0.15 mL / kg body weight of fleroxacin injection hydrochloride (product of our company), once a day, for two consecutive days.

[0098] Experimental results: On the 3rd day of medication, the mental state of the pig group improved and the feeding resumed; on the 7th day of medication, the morbidity and mortality were effectively controlled; on the 15th day of medication, the epidemic situation was completely controlled and there was no more death.

[0099] Clinical experiment two

[0100] Experimental site: In October 2022, a farm in Shangrao, Jiangxi, with 600 sows;

[0101] According to the feedback, the mortality rate of nursery pigs was 30%, the respiratory symptoms were obvious, the effect of antibiotic treatment was not good, the abortion rate of the sow group was 9%, and a series of reproductive problems such as a large number of dead fetuses, mummies, and return to estrus occurred.

[0102] Medication plan: (1) Pregnant sows: Add 2 kg of the traditional Chinese medicine composition prepared in Example 3 and 1 kg of 20% tylvalosin to each ton of feed, and use continuously for 7 days; (2) Commercial pigs (piglets, middle pigs, big pigs), replacement sows: Add 2 kg of the traditional Chinese medicine composition prepared in Example 3, 1 kg of 20% tilmicosin, and 1 kg of compound amoxicillin powder to each ton of feed, and use continuously for 7 days; (3) Nursing piglets with high mortality and diarrhea in the delivery room: Add 2 kg of Landian and 2 kg of Gankangshu (Yinzhi Jiedu Granules) to each ton of feed, and the sows are used from 14 days before delivery to 7 days after delivery.

[0103] Experimental results: After treating the breeding pig group with the above plan, the changes in the porcine reproductive and respiratory syndrome (PRRS) antibody of the pigs were detected, as shown in Figure 3 , Figure 4 , it can be seen that the coefficient of variation decreased from 55.2% to 25.9%; the S / P value was less than 2.5. This shows that the treatment method of the traditional Chinese medicine composition of the present invention effectively controls clinical problems such as abortion and stillbirth of sows, and the PRRS problem has been effectively controlled; it can quickly control the morbidity and mortality of the nursery pig group and make them gradually recover.

[0104] The above are only the preferred embodiments of the present invention, and do not impose any form of limitation on this application. Although this application is disclosed above with the preferred embodiments, it is not intended to limit this application. Any person skilled in the art, without departing from the scope of the technical solution of this application, making some changes or modifications using the technical content disclosed above is equivalent to equivalent implementation cases. However, as long as it does not depart from the content of the technical solution of the present invention, any simple modification, equivalent change and modification made to the above embodiments based on the technical essence of the present invention still fall within the scope of the technical solution.

Claims

1. A method for preparing a Chinese medicine composition for treating blue ear disease in pigs, characterized in that: The method comprises the following preparation steps: (1) mixing and crushing Chinese medicinal materials, adding fermentation bacteria thereto, and fermenting for 3-5 days to obtain a Chinese medicinal fermentation mixture; (2) adding complex enzyme to yeast powder, adjusting the pH to 6-9, performing enzymolysis at 40-60° C. for 3-5 hours, inactivating the enzyme, and drying to obtain yeast peptide liquid; (3) taking an ascorbic acid solution and a sodium selenite solution and mixing them evenly, adjusting the pH of the mixed system, adding the yeast peptide solution thereto, mixing evenly, then adding gelatin and mixing evenly, and drying to obtain a yeast peptide complex; (4) Evenly mixing the Chinese medicine fermentation mixture of step (1) and the yeast peptide complex of step (4) to prepare the Chinese medicine composition for treating porcine reproductive and respiratory syndrome.

2. The method for preparing the Chinese medicine composition according to claim 1, characterized in that: The Chinese medicinal materials in step (1) include, by weight, 10-20 parts of polygonum multiflorum, 5-15 parts of moutan bark, 10-30 parts of astragalus, 10-25 parts of pulsatilla, 1-10 parts of sophora flavescens, 1-15 parts of ephedra root, 20-30 parts of coix seed, 10-30 parts of liquorice, 5-20 parts of isatis indigotica, 1-10 parts of notopterygium wilfordii, 10-20 parts of chuanxiong, 5-15 parts of cassia twig, 10-25 parts of white peony root, 5-20 parts of dried ginger, 10-20 parts of siler, 5-25 parts of cyperus rotundus, 10-20 parts of stir-fried atractylodes, and 1-10 parts of codonopsis pilosula.

3. The method for preparing the Chinese medicine composition according to claim 1, characterized in that: The fermentation strain in step (1) is Bacillus coagulans.

4. The method for preparing the Chinese medicine composition according to claim 1, characterized in that: The complex enzyme in step (2) comprises papain and trypsin in a mass ratio of (2.5-3.5):

1.

5. The method for preparing the Chinese medicine composition according to claim 4, characterized in that: The dosage of the complex enzyme is 0.2-0.5wt% of the yeast powder.

6. The method for preparing the Chinese medicine composition according to claim 1, characterized in that: The molar ratio of ascorbic acid to sodium selenite in step (3) is (4-6):

1.

7. The method for preparing the Chinese medicine composition according to claim 1, characterized in that: The amount of gelatin used in step (3) is 5-10 wt % of the yeast peptide solution.

8. The method for preparing the Chinese medicine composition according to claim 1, characterized in that: The mass ratio of the traditional Chinese medicine fermentation mixture and the yeast peptide complex in step (4) is (2-3):

1.

9. A Chinese medicine composition for treating porcine blue ear disease prepared by the preparation method according to any one of claims 1 to 8.

10. The Chinese medicine composition according to claim 9, characterized in that: The usage amount of the traditional Chinese medicine composition is 0.1-0.5wt% of the feed used.

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