A traditional Chinese medicine composition for treating blue ear disease in pigs and preparation method thereof

Abstract

The present invention belongs to the field of drug preparation technology, and in particular to a Chinese medicine composition for treating blue ear disease in pigs and its preparation method. The preparation method of the Chinese medicine composition of the present invention comprises the following preparation steps: Chinese medicinal materials are mixed and crushed, fermentation strains are added thereto, and fermentation is carried out for 3-5 days to obtain a Chinese medicine fermentation mixture; a complex enzyme is added to yeast powder, and after adjusting the pH to 6-9, enzymolysis is carried out at 40-60 ° C for 3-5 hours, the enzyme is killed, and dried to obtain a yeast peptide liquid; a solution of ascorbic acid is mixed with a solution of sodium selenite, and after adjusting the pH of the mixed system, yeast peptide liquid is added thereto, mixed evenly, gelatin is added and mixed evenly, and dried to obtain a yeast peptide complex; the Chinese medicine fermentation mixture and the yeast peptide complex are mixed evenly to prepare the Chinese medicine composition for treating blue ear disease in pigs. The present invention provides a Chinese medicine and polypeptide formula with good efficacy for blue ear disease in pigs.

Description

Technical Field

[0001] The invention belongs to the technical field of drug preparation, and particularly relates to a traditional Chinese medicine composition for treating blue ear disease in pigs and a preparation method thereof. Background Art

[0002] Blue ear disease (PRRS) is the leading killer of pigs' immune systems. This problem has long plagued pig farmers, causing significant losses for many farms. Also known as porcine reproductive and respiratory syndrome (PRRS), PRRS has a significant impact on sows, piglets, weaned pigs, and adult pigs. Caused by the porcine reproductive and respiratory syndrome virus (PRRS) in the arteriviridae family, PRRS is characterized by reproductive failure, respiratory distress, blue-purple ear discoloration, immune decline, and the development of other diseases. Currently, the primary method for preventing PRRS is vaccination, but an ideal PRRS vaccine has yet to be developed, and the vaccine's protective ability and ability to prevent breakthrough infections still need improvement. Furthermore, there are currently no commercially available antiviral drugs directly targeting PRRS. There are only reports of PRRS being eradicated on pig farms using traditional Chinese medicine and antibiotics such as tilmicosin. While antibiotic treatment has some effectiveness, relapses occur quickly after discontinuation of the drug.

[0003] Patent CN 118903077 A discloses the use 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. The patent also discloses a composition of the drug against porcine reproductive and respiratory syndrome virus and PRRSV, which can be used to prevent PRRSV infection and / or treat PRRSV-induced porcine reproductive and respiratory syndrome. Although the invention tested the cell viability of Marc-145 cells at different shikonin concentrations, the PRRSV inhibition rate at different shikonin concentrations, and the results of animal serum antiviral tests, it lacks actual clinical trials on pigs and is still a long way from market launch. In addition, when using some traditional Chinese medicines to treat blue ear disease in pigs, traditional Chinese medicines are often directly added after being crushed. In actual application, this has disadvantages such as poor palatability, large addition amounts, and slow onset of effect, resulting in unsatisfactory therapeutic effects.

[0004] Therefore, developing a drug that can effectively treat porcine blue ear disease is of great significance to the breeding industry. Summary of the Invention

[0005] In response to the deficiencies in the prior art, the present invention provides a traditional Chinese medicine composition for treating blue ear disease in pigs and a preparation method thereof. The present invention discloses a formula of traditional Chinese medicine materials, and compounded polypeptide substances and optimized the formula. The prepared traditional Chinese medicine composition has good therapeutic effect on blue ear disease in pigs.

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

[0007] The first aspect of the present invention provides a method for preparing a traditional 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 enzymatic hydrolysis at 40-60°C for 3-5 hours, inactivating the enzyme, and drying to obtain yeast peptide liquid;

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

[0011] (4) The Chinese medicine fermentation mixture of step (1) and the yeast peptide complex of step (4) are evenly mixed 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 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.

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

[0014] In some embodiments, the complex 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] The second aspect of the present invention provides a traditional Chinese medicine composition for treating blue ear disease in pigs prepared by the preparation method described in the above scheme.

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

[0021] The present invention obtains a traditional Chinese medicine formula through repeated experiments, which has a good therapeutic effect on blue ear disease in pigs. The traditional Chinese medicine formula contains 18 kinds of medicinal materials, including bistort, peony bark, astragalus, pulsatilla, sophora flavescens, ephedra root, coix seed, liquorice, isatis indigotica, notopterygium wilfordii, Chuanxiong, cassia twig, white peony root, dried ginger, saposhnikovia divaricata, scutellaria baicalensis, stir-fried atractylodes, and codonopsis pilosula. The effects of each medicinal material are as follows: bistort: ​​clears heat and detoxifies, cools blood and stops bleeding, calms the liver and stops wind, eliminates carbuncle and disperses nodules; peony bark: clears heat and cools blood, promotes blood circulation and removes stasis, and reduces deficiency fever; astragalus: replenishes qi and raises yang, strengthens the body's defense system, and promotes diuresis and reduces swelling; pulsatilla: clears heat and detoxifies, cools blood, improves eyesight, and dries dampness; sophora flavescens clears heat and dries dampness, treats damp-heat diarrhea, bloody stool, and jaundice; ephedra root: strengthens the body's surface and stops sweating, promotes diuresis and reduces swelling, relieves cough and asthma; coix seed: promotes diuresis and promotes infiltration =Wetness, spleen and stomach tonifying and diarrhea, spleen deficiency diarrhea; Licorice: tonifies the spleen and replenishes qi, eliminates phlegm and relieves cough, harmonizes various medicines; Isatis indigotica: clears heat and detoxifies, cools blood and eliminates spots, relieves exterior pathogens; Notopterygium: dispels wind and cold, dispels rheumatism, and relieves pain; Chuanxiong: promotes blood circulation and qi, dispels wind and relieves pain, a blood-qi medicine; Cassia twig: induces sweating and relieves muscles, warms and unblocks the meridians, assists yang and transforms qi; White peony root: tonifies the kidney and strengthens the spleen, nourishes blood and regulates menstruation, and calms liver yang; Dry ginger: warms the middle and dispels cold, restores yang and unblocks the meridians, warms the lungs and transforms fluid; Saposhnikovia: pungent and sweet dispersing, dispels wind and relieves exterior pathogens, and relieves rheumatism; Coptis chinensis: cools blood and stops bleeding, clears heat and detoxifies; Stir-fried Atractylodes: invigorates the spleen and replenishes qi, dries dampness and promotes diuresis, tonifies qi and strengthens the spleen and stops diarrhea; Codonopsis pilosula: tonifies the middle and replenishes qi, promotes fluid and nourishes blood, moistens the lungs and produces fluid.

[0022] In order to further improve the efficacy of the medicine, the present invention uses Bacillus coagulans to ferment the Chinese medicinal materials. On the one hand, Bacillus coagulans can produce a variety of enzymes during the fermentation process, such as cellulase, amylase, and protease. These enzymes can decompose cellulose, starch, and protein in the mixed medicinal materials. For medicinal materials rich in polysaccharides and proteins such as astragalus and codonopsis, enzymatic hydrolysis can better dissolve the effective ingredients such as astragalus polysaccharides and codonopsis saponins, thereby 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, which improve the bitter taste of the mixed medicinal materials. For pigs, it can improve their acceptance of the drug and ensure that the pigs can take in enough drugs for the treatment of blue ear disease. In addition, during the fermentation process of Bacillus coagulans, organic acids such as lactic acid and butyric acid are produced. These organic acids can adjust the pH value of the fermentation system and create an acidic environment that is conducive to the stable existence of the drug ingredients. At the same time, organic acids also have certain antibacterial activity, which can inhibit the growth of harmful bacteria and provide protection for the intestinal health of pigs. Furthermore, these metabolites may interact with the ingredients in the medicinal materials to produce new pharmacologically active substances. The toxic substances in the fermented Chinese medicinal materials are relatively reduced, which can regulate the intestinal microecology of pigs, enhance their immune function, improve their physiological state, and enhance their disease resistance.

[0023] Because being ill may cause the absorption and utilization of Chinese medicine by the pig's stomach and intestines to decline, for this problem, the present invention uses compound enzyme (papain and trypsin) to enzymolyze yeast powder, papain mainly acts on the carboxyl side peptide bond of basic amino acid residues such as arginine and lysine, and trypsin specifically hydrolyzes the carboxyl side peptide bond of lysine and arginine. During the enzymolysis process, these enzymes cut the peptide bonds in the yeast protein, and the macromolecular yeast protein is gradually decomposed into smaller peptide segments. By regulating the pH and temperature, the enzymolysis reaction is promoted, and by controlling the enzymolysis time, the molecular weight distribution of the polypeptide is controlled, so that the obtained yeast peptide has good biological activity and absorbability. In order to improve the stability of yeast peptides, the present invention uses ascorbic acid and sodium selenite to modify the yeast peptides after enzymolysis under a certain pH and raw material ratio, wherein ascorbic acid can prevent selenium ions from being oxidized, promotes selenium ions to interact with the amino acid residues in the yeast peptides, forms complexes or chelates, and enhances the stability and biological activity of yeast peptides. After gelatin is added, functional groups such as amino and carboxyl groups in the gelatin molecules will undergo physical adsorption or chemical cross-linking reactions with yeast peptides to form a complex structure, and a stable yeast peptide complex will be obtained after drying.

[0024] The yeast peptide complex and the traditional Chinese medicine fermentation mixture are combined in appropriate proportions. Ingredients such as astragalus polysaccharides and codonopsis saponins in the traditional Chinese medicine fermentation mixture can activate the pig's immune system and promote the proliferation and differentiation of immune cells. The yeast peptides in the yeast peptide complex can provide nutritional support and enhance immune cell activity. The synergistic effect of the two can more effectively regulate the pig's immune system and enhance its resistance to the porcine reproductive and respiratory syndrome virus (PRRSV). Furthermore, the yeast peptides in the yeast peptide complex can promote intestinal absorption of the active ingredients of traditional Chinese medicine. For example, yeast peptides can regulate the function of intestinal mucosal cells and increase cell membrane permeability, allowing the active ingredients in the traditional Chinese medicine fermentation mixture, such as polysaccharides, saponins, and amino acids, to better enter the bloodstream and exert their therapeutic effects. Furthermore, ingredients such as gelatin in the yeast peptide complex can slowly release the yeast peptides and adsorbed traditional Chinese medicine ingredients in the gastrointestinal tract, prolonging their action time in the intestine, improving absorption efficiency, and maintaining the balance of the intestinal microbiome.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] 1. The present invention obtains a Chinese medicinal material formula through repeated experiments, and adds polypeptide substances thereto to optimize the formula and improve the therapeutic effect. The obtained Chinese medicinal composition has a good therapeutic effect on porcine blue ear disease.

[0027] 2. The present invention uses Bacillus coagulans to ferment the Chinese medicinal materials to better dissolve the active ingredients therein, improve the utilization rate of the medicinal materials, and improve the bitter taste of the mixed medicinal materials.

[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 yeast peptides after enzymatic hydrolysis to enhance the stability and biological activity of the yeast peptides and improve the efficacy. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 These are the results of molecular pathogen detection in pigs in clinical trial 1;

[0030] Figure 2 This is the result of the homology comparison between the site strain and the common strain in clinical experiment 1;

[0031] Figure 3 This is the blue ear antibody test result of pigs before treatment in clinical trial 2;

[0032] Figure 4 These are the results of blue ear antibody testing in pigs after treatment in Clinical Experiment 2. DETAILED DESCRIPTION

[0033] Various exemplary embodiments of the present invention are now described in detail. This detailed description should not be considered as a limitation of the present invention, but should be understood as a more detailed description of certain aspects, characteristics and embodiments of the present invention. It should be understood that the terms described in the present invention are only for describing particular embodiments and are not intended to limit the present invention. In addition, 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 smaller range between the intermediate value within any stated value or stated range and any other stated value or intermediate value within the range is also included in the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded within the scope.

[0034] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may be used in the practice or testing of the present invention. Various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention, as will be apparent to those skilled in the art. Other embodiments obtained from the present invention description will be apparent to those skilled in the art. This application description and examples are exemplary only.

[0035] The terms “include,” “including,” “have,” or “contain” used in this document are open-ended terms, meaning including but not limited to.

[0036] It should be noted that the operations of “grinding”, “centrifuging” and “stirring” described in the present invention are routine operations for those skilled in the art and can be selected according to actual operations.

[0037] The mass fractions described in the present invention may be fractions in grams, kilograms, tons or other mass units.

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

[0039] Example 1

[0040] A method for preparing a traditional Chinese medicine composition for treating blue ear disease in pigs, comprising the following preparation steps:

[0041] (1) Mixing Chinese medicinal materials, crushing them through a 60-mesh sieve, adding 1 wt% of Bacillus coagulans thereto, and fermenting them at 37° C. and 60% humidity for 5 days to obtain a Chinese medicinal 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 enzymatic hydrolysis at 40°C for 5 h, and then 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 uniformly, and after adjusting the pH of the mixed system to 7.5, 70 g of the yeast peptide solution stirred uniformly in step (1) was added thereto, mixed uniformly, and then 7 g of gelatin was added and mixed uniformly, 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 are calculated by weight and 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 cinnamon 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 enzymatic hydrolysis at 60°C for 3 h, and then 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) 30 g of the traditional Chinese medicine fermentation mixture of step (1) and 10 g of the yeast peptide complex of step (4) were mixed evenly to prepare the traditional Chinese medicine composition for treating porcine blue ear disease.

[0053] The Chinese medicinal materials used are calculated by weight and include 20 parts of polygonum multiflorum, 15 parts of peony bark, 30 parts of astragalus, 25 parts of pulsatilla, 10 parts of sophora flavescens, 15 parts of ephedra root, 30 parts of coix seed, 30 parts of liquorice, 20 parts of isatis indigotica, 10 parts of notopterygium wilfordii, 20 parts of chuanxiong, 15 parts of cinnamon twig, 25 parts of white peony root, 20 parts of dried ginger, 20 parts of siler, 25 parts of mianma guanzhong, 20 parts of stir-fried atractylodes, and 10 parts of codonopsis pilosula.

[0054] The complex enzyme used contained papain and trypsin in a mass ratio of 3:1.

[0055] Example 3

[0056] A method for preparing a traditional Chinese medicine composition for treating blue ear disease in pigs, comprising the following preparation steps:

[0057] (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;

[0058] (2) adding 0.35 wt% of complex enzyme and 4 times the weight of pure water to yeast powder, adjusting the pH to 8, performing enzymatic hydrolysis at 50°C for 2 h, and then inactivating the enzyme at 100°C for 10 min to obtain yeast peptide liquid;

[0059] (3) 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 the yeast peptide solution stirred uniformly in step (1) 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;

[0060] (4) 25 g of the traditional Chinese medicine fermentation mixture of step (1) and 10 g of the yeast peptide complex of step (4) were mixed evenly to prepare the traditional Chinese medicine composition for treating porcine blue ear disease.

[0061] The Chinese medicinal materials used are calculated by weight and 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 wilfordii, 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 mianma guanzhong, 15 parts of stir-fried atractylodes, and 5 parts of codonopsis pilosula.

[0062] The complex enzyme used contained papain and trypsin in a mass ratio of 2.5:1.

[0063] Example 4

[0064] A method for preparing a traditional Chinese medicine composition for treating blue ear disease in pigs, 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 blue ear disease in pigs, the specific implementation method is the same as that of Example 3, except that the complex enzyme in step (2) contains papain and trypsin in a mass ratio of 4:1.

[0067] Comparative Example 1

[0068] A method for preparing a traditional Chinese medicine composition for treating blue ear disease in pigs, the specific implementation method is the same as that of Example 3, except that step (1) is: mixing traditional Chinese medicine materials, crushing them through an 80-mesh sieve, and obtaining a traditional 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 mixture to 8, 140 g of a 20 wt% yeast powder solution dispersed in pure water was added and mixed uniformly. 10 g of gelatin was then added and mixed uniformly, and the mixture was 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) were mixed evenly to prepare the traditional Chinese medicine composition for treating porcine blue ear disease.

[0074] The Chinese medicinal materials used are calculated by weight and 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 wilfordii, 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 mianma guanzhong, 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 method for preparing a traditional Chinese medicine composition for treating blue ear disease in pigs, the specific implementation method is the same as that of Example 3, except that step (3) is: 100 mL of a 2 mol / L ascorbic acid aqueous solution and 100 mL of a 0.4 mol / L sodium selenite aqueous solution are mixed evenly, the pH of the mixed system is adjusted to 8, and 140 g of the yeast peptide solution stirred evenly in step (1) is added thereto, mixed evenly, and spray-dried to obtain a yeast peptide complex.

[0078] Comparative Example 4

[0079] A method for preparing a traditional Chinese medicine composition for treating blue ear disease in pigs, the specific implementation method 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 method for preparing a traditional Chinese medicine composition for treating blue ear disease in pigs, the specific implementation method is the same as that of Example 3, except that an equal mass of trypsin is used instead of the complex enzyme.

[0082] Comparative Example 6

[0083] A method for preparing a traditional Chinese medicine composition for treating blue ear disease in pigs, comprising the following preparation steps:

[0084] The Chinese medicinal materials are mixed, crushed and passed through an 80-mesh sieve to obtain a Chinese medicinal composition.

[0085] Performance Testing

[0086] 1. Antiviral experiments

[0087] Take equal weight of the Chinese medicine mixture / Chinese medicine composition obtained in the examples and comparative examples, add pure water to cover the medicinal materials of each group, heat and extract at 70°C for 0.5h, filter, repeat the extraction step twice, combine the extracts, concentrate and dry, and then prepare a standard solution with a concentration of 15 mg / mL with pure water.

[0088] Antiviral experiments were conducted according to patent CN 118903077 A: Marc-145 cells were cultured at a rate of 1.5×10 4Cells / well (volume 100 μL) were inoculated in a 96-well cell culture plate. When the cells adhered and the confluence rate exceeded 90%, they were replaced with DMEM maintenance medium (DMEM medium, 2% serum, penicillin 100 U / mL, streptomycin 100 μg / mL). Each infection group was added with 0.1 MOI (multiplicity of infection) of WUH3 porcine reproductive and respiratory syndrome virus. At the same time, each test solution was diluted into 6 concentration gradients with maintenance medium and added to each drug-treated well (Example 1-5, Comparative Example 1-6). Each concentration gradient had 3 replicates, and the final concentrations were 10 mg / mL, 5 mg / mL, 2.5 mg / mL, 1 mg / mL, and 0.1 mg / mL, respectively. Cultured in a 37°C cell culture incubator for 60 h. Take out the culture plate and freeze-thaw once at -20°C, blow each well and draw 180 μL into the viral nucleic acid purification kit machine to automatically extract RNA.

[0089] For other operations, please refer to patent CN 118903077 A

[0046] -

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

[0090] Table 1

[0091]

[0092] As shown in Table 1, the Chinese medicine mixtures prepared in Examples 1-3 have a good inhibitory effect on porcine reproductive and respiratory syndrome virus. Their IC 50 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 the traditional Chinese medicine components, making IC 50 The results show that the peptide molecular weight of the peptides in Comparative Example 2 is different from that in Comparative Example 5, Comparative Examples 4 and 5. The peptide molecular weight of the peptides in Comparative Example 1 is different from that in Comparative Example 6, which contains only crushed herbs. The efficacy of the Chinese medicine compositions in Comparative Example 1 is weaker than that in Comparative Example 3. 50 Significantly increased.

[0093] 2. Clinical trials

[0094] Clinical Trial 1

[0095] Experimental site: In mid-April 2023, a large-scale farm in Haozhou, Anhui Province, with a sow inventory of 530 and a fattening farm (3,000), with 600 pigs weighing about 30 kg in the affected pen;

[0096] According to feedback, sporadic deaths began to occur in mid-March, and multiple treatments were used during this period with poor results, and the morbidity and mortality rates showed an upward trend. Figure 1 、 2 .

[0097] Medication regimen: (1) Mix feed for the whole herd: add 3 kg of the Chinese herbal composition prepared in Example 3, 1 kg of Angel, and 1 kg of 20% tilmicosin per ton of feed, and use it continuously for 15 days; (2) Isolate sick pig pens: Add medicine to drinking water, add 100 g of the Chinese herbal composition prepared in Example 3, 100 g of 20% tilmicosin, and 100 g of Angel per 100 catties of water; (3) Intramuscular injection of seriously ill pigs: 0.15 mL / kg body weight of difloxacin hydrochloride injection (product of our company), once a day, for two consecutive days.

[0098] Experimental results: After 3 days of medication, the mental state of the pigs improved and feeding resumed; after 7 days of medication, the incidence and mortality rates were effectively controlled; after 15 days of medication, the epidemic was completely under control and no more deaths occurred.

[0099] Clinical Trial 2

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

[0101] According to feedback, the mortality rate of nursery pigs was 30%, respiratory symptoms were obvious, antibiotic treatment was ineffective, the abortion rate of sows was 9%, and there were a large number of stillbirths, mummies, estrus and other reproductive problems.

[0102] Dosage regimen: (1) Pregnant sows: add 2 kg of the Chinese medicine composition prepared in Example 3 and 1 kg of 20% tyvalosin per ton of feed, and use it continuously for 7 days; (2) Commercial pigs (piglet, medium pig, large pig) and gilts: add 2 kg of the Chinese medicine composition prepared in Example 3, 1 kg of 20% tilmicosin and 1 kg of compound amoxicillin powder per ton of feed, and use it continuously for 7 days; (3) Suckling piglets with high mortality and diarrhea in the delivery room: add 2 kg of Lan Dian and 2 kg of Gan Kang Shu (Yinzhi Jiedu Granules) per ton of feed, and use it continuously for sows from 14 days before delivery to 7 days after delivery.

[0103] Experimental results: After the breeding pigs were treated with the above drug regimen, the changes in the blue ear antibodies of the pigs were detected. Figure 3 、 Figure 4 As can be seen, the dispersion decreased from 55.2% to 25.9%, and the S / P values ​​were all less than 2.5. This indicates that the treatment method of the Chinese herbal composition of the present invention effectively controls clinical problems such as miscarriage and stillbirth in sows, effectively controls blue ear disease, and can quickly control the morbidity and mortality of nursery pigs, leading to their gradual recovery.

[0104] The above is only a preferred embodiment of the present invention and does not limit the present application in any form. Although the present application is disclosed as above with a preferred embodiment, it is not intended to limit the present application. Any technician familiar with this profession, without departing from the scope of the technical solution of the present application, makes some changes or modifications using the technical content disclosed above, which are equivalent to equivalent implementation cases. However, any simple modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solution of the present invention are still within the scope of the technical solution.

Claims

1. A method for preparing a traditional 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) Add complex enzyme to yeast powder, adjust the pH to 6-9, perform enzymolysis at 40-60°C for 3-5 hours, inactivate the enzyme, and dry to obtain yeast peptide liquid; (3) Mixing the ascorbic acid solution and the sodium selenite solution evenly, adjusting the pH of the mixture, adding the yeast peptide solution, mixing evenly, then adding gelatin, mixing evenly, and drying to obtain a yeast peptide complex; (4) uniformly mixing the Chinese medicine fermentation mixture of step (1) and the yeast peptide complex of step (3) to prepare the Chinese medicine composition for treating porcine blue ear disease; The Chinese medicinal materials in step (1) are composed of 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; The fermentation strain in step (1) is Bacillus coagulans; The complex enzyme in step (2) comprises papain and trypsin in a mass ratio of (2.5-3.5):1; The molar ratio of ascorbic acid to sodium selenite in step (3) is (4-6):

1.

2. The method for preparing the Chinese medicine composition according to claim 1, wherein: The dosage of the complex enzyme is 0.2-0.5 wt % of the yeast powder.

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

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

1.

5. A traditional Chinese medicine composition for treating porcine blue ear disease prepared by the preparation method according to any one of claims 1 to 4.

6. The Chinese medicine composition according to claim 5, 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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