Medicine for improving immunity of bred animals

Abstract

The invention belongs to the field of veterinary drugs, and particularly relates to a medicine for improving immunity of bred animals. The medicine provided by the invention comprises the following active ingredients in parts by weight: 10-20 parts of withered grass 37 peptide and extracts of the following traditional Chinese medicinal materials: 15-30 parts of wintercreeper, 15-30 parts of astragalus membranaceus, 15-30 parts of spirulina, 10-20 parts of silybum marianum and 10-20 parts of herba epimedii. Wherein the withered grass 37 peptide is a biological active peptide with an anti-infection function, is composed of a plurality of amino acids and has broad-spectrum antibacterial activity. The medicine provided by the invention has the functions of inhibiting and sterilizing, regulating immunity and regulating body functions so as to improve the immunity of bred animals, thereby turning pathogenesis and ensuring the health of the bred animals. A curative effect experiment shows that the medicine can improve the growth performance of bred animals and effectively reduce breeding loss, has the advantages of being environmentally friendly, safe and capable of preventing diseases, and has a good market prospect.

Description

Technical Field

[0001] The invention belongs to the field of veterinary medicines, and in particular relates to a medicine for improving the immunity of farmed animals. Background Art

[0002] With the rapid development of the intensification and scale of the breeding industry, the expectations of breeding entities for animal production performance and return on investment are getting higher and higher. Disease is an important factor affecting whether production performance can be performed normally, and the level of animal immunity determines the incidence rate. In today's breeding industry, affected by breeding methods, feeding methods and unreasonable breed improvement, farmed animals are in a state of stress, manifested as low immunity or even immunosuppression, causing significant losses to the breeding industry.

[0003] The manifestations of low immunity in poultry are as follows: the majority of the flock is in good spirits, but some of them suffer sporadic casualties, and their bodies are thin, etc. Individuals develop unevenly, and the absorption rate of nutrients is low. The resistance to viruses and bacteria is reduced, and they are easy to be infected with diseases.

[0004] Signs of low immunity in pigs include loss of appetite, listlessness, rough hair, and diarrhea in piglets. If pigs have a loss of appetite or even refuse to eat, it means they may be unwell or infected with respiratory diseases or other viruses. If pigs often curl up and are unwilling to get up and move, or even have abnormal symptoms such as convulsions, it means they are in a more serious state of discomfort. If the hair of pigs becomes dry and yellow or rough, it also means they may have health problems.

[0005] Low immunity in fish is manifested by uneven individual development and increased incidence of diseases. Fish have intestinal disorders, severe congestion, and low feed utilization. Low immunity reduces resistance to bacteria, viruses, and parasites, making them susceptible to infection and difficult to cure. Fish have pale body color, short and fat body, and reduced meat quality. They have low stress resistance, are prone to unexplained death, and have reduced feeding ability.

[0006] Immunosuppression is a temporary or permanent disorder of the body's immune response. It is caused by damage to the immune system and leads to increased susceptibility to disease, thus often resulting in a significant decrease in production performance. The lack of certain nutrients can directly affect the development and function of the immune system. Immunosuppression has become a common and harmful disease. Its causes are relatively complex and may involve multiple aspects such as mycotoxin contamination of feed, unbalanced feed nutrition, poor feeding conditions, viral and bacterial infections, etc.

[0007] Immunosuppression can lead to slow growth, reduced feed utilization, and increased mortality in animals, causing serious economic losses to the animal husbandry industry. Therefore, taking effective prevention and treatment measures is of great significance to ensure the healthy development of the animal husbandry industry. Summary of the invention

[0008] The purpose of the present invention is to provide a medicine for improving the immunity of farmed animals.

[0009] Based on the above purpose, the present invention adopts the following technical solutions:

[0010] The present invention provides a medicine for improving the immunity of farmed animals, wherein the active ingredient of the medicine is composed of 10 to 20 parts by weight of subtilisin and extracts of the following Chinese medicinal materials: 15 to 30 parts by weight of Euonymus fortunei, 15 to 30 parts by weight of Astragalus, 15 to 30 parts by weight of Spirulina, 10 to 20 parts by weight of Silybum marianum, and 10 to 20 parts by weight of Epimedium, wherein the Chinese medicinal materials are calculated in terms of crude drug amount. The subtilisin is a type of bioactive peptide with anti-infection function, which is composed of multiple amino acids and has broad-spectrum antibacterial activity.

[0011] Preferably, the medicine comprises, by weight, 20 parts of heptacopeptide subtilisin, 30 parts of euphorbia fargesii, 20 parts of astragalus, 20 parts of spirulina, 10 parts of silybum marianum and 10 parts of epimedium.

[0012] Preferably, the dosage form of the drug is an oral dosage form.

[0013] Preferably, the oral dosage form is powder, granules or oral solution.

[0014] Preferably, the extract of the Chinese medicinal material is prepared by the following steps:

[0015] The Chinese medicinal materials of Euonymus fortunei, Astragalus, Spirulina, Milk thistle and Epimedium are mixed and crushed, and then an ethanol solution is added for extraction, and filtered to obtain a medicinal residue and an ethanol crude extract;

[0016] Add water to the medicinal residue to extract, filter, and obtain a water extract;

[0017] The ethanol in the crude ethanol extract is recovered, and the crude ethanol extract after the ethanol is recovered and the water extract are combined, and the filtrate is concentrated to 1 to 2 times the weight of the Chinese medicinal material, so as to obtain the extract of the Chinese medicinal material.

[0018] Preferably, the volume concentration of the ethanol solution is 60% to 80%, and during alcohol extraction, the total amount of Chinese medicinal materials and the ethanol solution are added in a weight ratio of 1: (8 to 12).

[0019] Preferably, the alcohol extraction is carried out by reflux at a reflux temperature of 80°C to 90°C.

[0020] Preferably, during water extraction, the total amount of Chinese medicinal materials and water are added in a weight ratio of 1: (8-12).

[0021] Preferably, the membrane filtration is ultrafiltration separation, the material is polyethersulfone or polypropylene, and the pore size is 0.45 μm to 1.0 μm.

[0022] Preferably, the drug is in the form of an oral liquid, which is prepared by adding heptapeptide subtilisin and a preservative to the extract of the traditional Chinese medicine and mixing well.

[0023] Preferably, the drug is a powder or granule, and the extract of the Chinese medicinal material is spray-dried and then added with Trisaccharide Subtilisin, and then the excipients and preservatives are mixed evenly to obtain a powder; or the extract of the Chinese medicinal material is spray-dried and then added with Trisaccharide Subtilisin, and then excipients, preservatives, and adhesives are added, and then granulated, dried, and granulated to obtain granules.

[0024] Preferably, the excipient is sucrose and / or maltodextrin; the preservative is ethyl hydroxybenzoate; and the binder is starch slurry.

[0025] The heptapeptide of subtilisin in the present invention and the Chinese medicinal materials used to obtain the extract can be purchased from the market. The formulation principle of the medicine is as follows:

[0026] Euonymus fortunei: An evergreen vine shrub of the genus Euonymus of the family Celastraceae. The medicinal part is the stems and branches with leaves. It tastes bitter and is warm in nature. It enters the liver and kidney meridians. It contains triterpenes, flavonoids, lignin, steroids and phenolic acids. It has the effects of enhancing immunity, relaxing muscles and tendons, stopping bleeding and eliminating cancer.

[0027] Astragalus: A perennial herb of the genus Astragalus in the leguminous family, the medicinal part is the root; it tastes bitter, pungent, sweet, and slightly warm in nature, and enters the spleen and lung meridians; it contains saponins, flavonoids, amino acids, and polysaccharides; it has the effects of tonifying Qi and raising Yang, consolidating the exterior and stopping sweating, promoting diuresis and reducing swelling, promoting fluid and nourishing blood, relieving stagnation and relieving numbness, expelling toxins and discharging pus, and astringing sores and promoting tissue regeneration; it is mainly used to treat Qi deficiency and fatigue, sinking of Qi in the middle, internal heat and thirst, strengthening the Wei and consolidating the exterior, expelling toxins and promoting tissue regeneration.

[0028] Spirulina: The algae of various spirulina species, including Spirulina platensis, of the genus Spirulina in the family Oscillatoriaceae; it tastes bitter, salty, and cold in nature, and enters the liver, stomach, and kidney meridians; it is high in protein, low in fat, and low in sugar, and contains a variety of vitamins and trace elements. In addition, the active ingredients in Spirulina, such as phycocyanin (CPC), algal polysaccharide (PSP), γ-linolenic acid methyl ester (GLAME), β-carotene, chlorophyll a, etc., have a regulatory effect on many functions of animals; it has the effect of enhancing immunity.

[0029] Milk thistle: the dried mature fruit of the Asteraceae plant Milk thistle; bitter, cool, and enters the liver and gallbladder meridians; contains silymarin, fumaric acid, and flavonoids; has the functions of clearing away heat and detoxifying, soothing the liver and promoting bile secretion; used for damp-heat in the liver and gallbladder.

[0030] Epimedium: The dried leaves of Epimedium, Epimedium sagittatum, Epimedium pubescentis or Epimedium koreana of the Berberidaceae family are pungent, sweet, warm, and enter the liver and kidney meridians; contain flavonoids, polysaccharides, lignans, alkaloids, volatile oils, trace elements, essential fatty acids, phytosterols, cetyl alcohol and vitamins; have the effects of tonifying kidney yang, strengthening muscles and bones, and removing rheumatism; used for kidney yang deficiency, enhancing immunity, etc.

[0031] Subtilisin 37 peptide: It is a light yellow powder with no odor. It is composed of 37 amino acid residues and its molecular formula is C 162 H 254 N 50 O 51 S5, molecular weight 3.8KD, two disulfide bonds connect the two α-helices it forms to form a tight structure, super stable in nature, and has good resistance to high temperature, acid, alkali, pepsin, and trypsin. Subtilisin mainly works in the intestine and will not enter the blood in its intact form. Antimicrobial peptides are a class of bioactive peptides with anti-infection function. They are composed of amino acids, widely distributed in organisms, have broad-spectrum antibacterial activity, and are part of the innate immune defense system of organisms. They mainly play an anti-infection role by directly inhibiting or killing pathogens and regulating the body's immune function. Compared with antibiotics, antimicrobial peptides are not easy to develop drug resistance during use due to their unique mechanism of action (membrane perforation), and they are green and safe without residue.

[0032] In the scheme provided by the present invention, Eucommia ulmoides and Astragalus membranaceus are sweet and warm in nature, and have the functions of improving immunity, replenishing qi and raising yang, and nourishing qi and blood as the main medicines; Spirulina and Milk Thistle are bitter and cold in nature and are the ministerial medicines, which have the functions of regulating immunity, clearing away heat and detoxifying, and soothing the liver and promoting bile secretion; Epimedium brevifolia is used as an adjuvant to replenish kidney yang and strengthen tendons and bones, and Herba Subtilisin 37 peptides directly inhibit or kill pathogenic bacteria and regulate the body's immune function to exert anti-infection effects in two ways. The combination of all the medicines can play the functions of replenishing qi and nourishing blood, soothing the liver and promoting bile secretion, clearing away heat and detoxifying, inhibiting sterilization, regulating immunity, regulating body functions and improving immunity, thereby reversing the pathogenesis and interrupting the disease. The prepared product can regulate immunity from multiple directions and multiple targets, thereby restoring the body.

[0033] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0034] The drug provided by the present invention has the functions of inhibiting sterilization, regulating immunity, regulating body functions and improving immunity, thereby reversing the pathogenesis and ensuring the health of animals. The efficacy experiment shows that the drug of the present invention can improve the growth performance of animals and effectively reduce breeding losses; it is also green and safe, and can prevent diseases, and has a good market prospect. DETAILED DESCRIPTION

[0035] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below through specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0036] Example 1

[0037] A medicine for improving the immunity of farmed animals, wherein the active ingredients of the medicine are composed of 20 parts by weight of heptapeptide-30 of subtilisin and extracts of the following Chinese medicinal materials: 30 parts by weight of Euonymus fortunei, 20 parts by weight of Astragalus membranaceus, 20 parts by weight of Spirulina, 10 parts by weight of Silybum marianum, and 10 parts by weight of Epimedium, wherein the Chinese medicinal materials are calculated in terms of raw drug amount.

[0038] The extract of Chinese medicinal materials is prepared by the following steps:

[0039] A. Weigh euonymus, astragalus, spirulina, milk thistle and epimedium according to the above weight ratio, mix and crush these Chinese medicinal materials, and then add 70% ethanol solution by volume for reflux extraction, the reflux temperature is 85±5°C, filter after the reflux extraction, add ethanol solution again for reflux extraction, extract 3 times in total, each extraction is 1.5h, and the total amount of Chinese medicinal materials and ethanol solution are added in a weight ratio of 1:10 for each extraction; combine the ethanol extracts to obtain a crude ethanol extract, and the residues enter the next step for water extraction;

[0040] B. Add water to the residue obtained in step A for decoction extraction, extract 3 times, each time for 1 hour, and feed the total amount of Chinese medicinal materials and water at a weight ratio of 1:10 each time; filter, combine the filtrates of the three times to obtain a water extract;

[0041] C. Recovering ethanol from the crude ethanol extract, combining the crude ethanol extract after ethanol recovery and the water extract obtained in step B, filtering with a membrane (made of polyethersulfone with a pore size of 0.45 μm to 1.0 μm), and concentrating the filtrate to 1 to 2 times the weight of the Chinese medicinal materials, thereby obtaining an extract of the Chinese medicinal materials.

[0042] Furthermore, 20 parts by weight of heptapeptide-30 of subtilisin are added to the extract prepared in step C, and 0.5% of the total weight of the extract is added with ethyl hydroxybenzoate (preservative), and the mixture is mixed to obtain an oral liquid dosage form.

[0043] Alternatively, the extract obtained in step C is first spray-dried, and then 20 parts by weight of heptapeptide-37 of subtilisin is added and mixed to obtain a powdered active ingredient; maltodextrin (excipient) is weighed at 9 times the total weight of the active ingredient, and ethyl hydroxybenzoate (preservative) is weighed at 5% of the total weight of the active ingredient, the preservative and excipient are first mixed in equal amounts, and then the powder is added and mixed evenly to obtain a powder.

[0044] As another embodiment, starch slurry (binder) is added to the powder according to 10% of the weight of the powder, mixed and granulated with a 20-mesh sieve, dried at 80±5° C., and sieved with 18 and 80 mesh sieves to obtain granules.

[0045] Example 2

[0046] The content of a medicine for enhancing the immunity of farmed animals is basically the same as that of Example 1, except that the active ingredient of the medicine consists of 10 parts by weight of heptapeptide-37 from subtilisin and extracts of the following Chinese medicinal materials: 20 parts by weight of Euonymus fortunei, 30 parts by weight of Astragalus membranaceus, 15 parts by weight of Spirulina, 15 parts by weight of Silybum marianum, and 15 parts by weight of Epimedium.

[0047] Comparative Example 1

[0048] The content of a medicine is substantially the same as that of Example 1, except that the medicine contains, by weight, 30 parts of Eucommia ulmoides, 30 parts of Astragalus membranaceus and 20 parts of Spirulina.

[0049] Comparative Example 2

[0050] The content of a medicine is substantially the same as that of Example 1, except that: the medicine contains 20 parts of milk thistle and 20 parts of epimedium by weight.

[0051] Comparative Example 3

[0052] The content of a medicine is basically the same as that of Example 1, except that: the medicine contains 10 parts of heptacopeptide subtilisin by weight.

[0053] The efficacy verification experiment of the drug of the present invention for effectively improving immunity:

[0054] In order to verify the efficacy of the drug of the present invention in effectively improving immunity, the drugs prepared in Examples 1 and 2 of the present invention were used as examples to conduct efficacy verification experiments. At the same time, a blank group and 4 control groups were set up. The control groups took the drugs of Comparative Examples 1 to 3 respectively, and the commercially available drug Astragalus polysaccharide powder group, specification: each 1g contains no less than 450mg of Astragalus polysaccharide, batch number: 92011191M. Astragalus polysaccharide powder, the function and indications are to invigorate qi and strengthen the body's resistance. It is used for the preventive treatment of infectious bursal disease in chickens; to increase the antibody level of swine fever and swine foot-and-mouth disease vaccines; to improve the nonspecific immune function of aquatic animals.

[0055] Usage and Dosage: For chickens, mixed feed, 200 mg per 1 kg of feed, free feeding, for 5 to 7 days; for pigs, mixed feed, 200 mg per 1 kg of feed, free feeding, administration 3 days before vaccination, for 30 days; for grass carp, tilapia, grouper, Chinese soft-shelled turtle, mixed feed, add 400 mg per 1 kg of feed, feeding continuously for 30 days.

[0056] Table 1 Medication status of each group

[0057]

[0058] 1. Target animal for drug efficacy evaluation experiment: broiler chickens:

[0059] 1.1 Experimental subjects

[0060] A certain animal husbandry company in Xishui County, Zunyi City, Guizhou Province, selected 700 5-day-old double A large broilers from the same batch with no significant difference in initial body weight, and randomly divided them into 7 groups, with 100 chickens in each group. They used broiler feed produced by Wellcome Animal Husbandry, and had free access to food and drinking water.

[0061] 1.2 Experimental drugs

[0062] The experimental drugs are the drugs prepared in Examples 1-2 of the present invention, the drugs prepared in Comparative Examples 1-3, and Astragalus polysaccharide powder. The dosage of each group is: centralized administration in the morning, mixed feeding, adding 200 mg of the drug to every 1 kg of feed, free feeding, and continuous use for 7 days.

[0063] The blank group was only given the same drinking water and feed.

[0064] 1.3 Measurement indicators and measurement methods

[0065] 1.3.1 Growth indicators

[0066] The body weight of each group of broilers was measured on an empty stomach in the morning on the first and eighth days of the experiment, and the average daily weight gain of each group of broilers was calculated.

[0067] 1.3.2 Serum immune indicators

[0068] On the 8th day of the experiment, 10 broilers were randomly selected from each group under fasting conditions in the morning, blood was collected from the wing vein, centrifuged at high speed, and the serum was separated and frozen for storage. IgA, IgG, IgM and TNF-α were determined according to the operating procedures of the immunoassay kit.

[0069] 1.3.3 Organ index determination

[0070] After blood collection, the broiler chickens were dissected, and the liver, spleen, and bursa of Fabricius were taken out and weighed to calculate the organ index. Liver index = liver weight mg / chicken individual weight g; spleen index = spleen weight mg / chicken individual weight g; bursa index = bursa weight mg / chicken individual weight g.

[0071] 1.4 Measurement results

[0072] 1.4.1 Growth index results are shown in Table 2.

[0073] Table 2 Statistics of growth index results of each group

[0074]

[0075] The experimental results in Table 2 show that compared with the blank group, the average daily weight gain of Example 1 and Example 2 increased significantly, the Comparative Example 1, Comparative Example 3 and the Astragalus polysaccharide powder group increased slightly, and there was no significant difference between Comparative Example 2 and the blank group.

[0076] 1.4.2 Serum immune index results are shown in Table 3.

[0077] Table 3 Statistics of nonspecific immunity results of each group

[0078] Group drug IgA (mg / mL) IgG (mg / mL) IgM (mg / mL) TNF-α(pg / mL) Experimental Group 1 Example 1 <![CDATA[1.84±0.07 a ]]> 4.85±0.23 2.49±0.08 <![CDATA[9.21±0.56 c ]]> Experimental Group 2 Example 2 <![CDATA[1.82±0.06 a ]]> 4.83±0.24 2.46±0.07 <![CDATA[9.29±0.52 c ]]> Experimental Group 3 Comparative Example 1 <![CDATA[1.80±0.04 a ]]> 4.87±0.28 2.33±0.08 <![CDATA[9.58±0.73 bc ]]> Experimental Group 4 Comparative Example 2 1.64±0.05c 4.86±0.26 2.06±0.04 <![CDATA[10.16±0.69 b ]]> Experimental Group 5 Comparative Example 3 <![CDATA[1.75±0.07 b ]]> 4.83±0.19 2.25±0.06 <![CDATA[9.75±0.68 b ]]> Experimental Group 6 Astragalus polysaccharide powder <![CDATA[1.73±0.06 b ]]> 4.81±0.32 2.21±0.04 <![CDATA[9.87±0.71 b ]]> Blank Group - <![CDATA[1.62±0.05 c ]]> 4.85±0.21 1.92±0.05 <![CDATA[10.98±0.86 a ]]>

[0079] The experimental results in Table 3 show that compared with the blank group, Example 1, Example 2 and Comparative Example 1 can all significantly increase the IgA content in broiler serum, followed by Comparative Example 3 and the Astragalus polysaccharide powder group, and there is little difference between Comparative Example 2 and the blank group; compared with the blank group, each experimental group can significantly reduce the TNF-α content in broiler serum, among which Example 1 is the lowest; for the values ​​of IgG and IgM, there is no significant difference between the experimental groups and the blank group.

[0080] 1.4.3 Organ index results are shown in Table 4.

[0081] Table 4 Statistics of organ index results of each group

[0082] Group drug Liver index Spleen index Bursal index Experimental Group 1 Example 1 <![CDATA[37.23±0.35 a ]]> 1.04±0.14 2.52±0.18 Experimental Group 2 Example 2 <![CDATA[37.12±0.38 a ]]> 1.07±0.16 2.51±0.19 Experimental Group 3 Comparative Example 1 <![CDATA[36.71±0.41 a ]]> 1.02±0.25 2.55±0.22 Experimental Group 4 Comparative Example 2 <![CDATA[35.67±0.32 b ]]> 1.06±0.17 2.52±0.17 Experimental Group 5 Comparative Example 3 <![CDATA[34.97±0.29 b ]]> 1.01±0.13 2.56±0.23 Experimental Group 6 Astragalus polysaccharide powder <![CDATA[35.21±0.26 b ]]> 1.05±0.21 2.53±0.19 Blank Group - <![CDATA[32.75±0.31 c ]]> 1.08±0.15 2.58±0.22

[0083] The experimental results in Table 4 show that, compared with the blank group, each experimental group can significantly improve the liver index of broiler chickens, among which Example 1 and Example 2 are the best; for spleen index and bursa of Fabricius index, there is no significant difference between the experimental groups and the blank group.

[0084] 2. Target animal for drug efficacy evaluation experiment: Pigs:

[0085] 2.1 Experimental subjects

[0086] An ecological agriculture company in Baidian Town, Anhui Province, selected 140 weaned piglets from the same batch with no significant difference in initial weight for the experiment. They were randomly divided into 7 groups, with 20 pigs in each group. They used pig feed produced by Kaifeng Zhengda Co., Ltd. and were allowed to eat and drink freely.

[0087] 2.2 Experimental drugs

[0088] The experimental drugs are the drugs prepared in Examples 1-2 of the present invention, the drugs prepared in Comparative Examples 1-3, and Astragalus polysaccharide powder. The dosage of each group is: centralized administration in the morning, mixed feeding, adding 200 mg of the drug to every 1 kg of feed, free feeding, and continuous use for 30 days.

[0089] The blank group was only given the same drinking water and feed.

[0090] 2.3 Measurement indicators and measurement methods

[0091] 2.3.1 Growth indicators

[0092] The body weight of weaned piglets in each group was measured on an empty stomach on the first and 31st day of the experiment, and the average daily weight gain of weaned piglets in each group was calculated.

[0093] 2.3.2 Diarrhea rate

[0094] The anus of each group of weaned piglets was checked regularly in the morning and afternoon every day, the diarrhea situation of the weaned piglets was recorded, and the diarrhea rate was calculated as (number of weaned piglets with diarrhea × number of diarrhea days) / (total number of weaned piglets × number of experimental days).

[0095] 2.3.3 Serum immune indexes

[0096] After the experiment, blood was collected from each group of weaned piglets through the jugular vein under fasting conditions, centrifuged at high speed, and the serum was separated and frozen for storage. LZM, IgA, IgG, IgM and TNF-α were determined according to the operating procedures of the immunoassay kit.

[0097] 2.4 Measurement results

[0098] 2.4.1 Growth index results are shown in Table 5.

[0099] Table 5 Statistics of growth index results of each group

[0100] Group drug Initial weight (kg) Final weight(kg) Average daily weight gain (g) Diarrhea rate (%) Experimental Group 1 Example 1 7.63±0.21 15.23±0.22 <![CDATA[253.33±9.51 a ]]> <![CDATA[1.54±0.13 d ]]> Experimental Group 2 Example 2 7.65±0.32 15.11±0.28 <![CDATA[248.67±9.32 a ]]> <![CDATA[1.97±0.22 c ]]> Experimental Group 3 Comparative Example 1 7.58±0.28 14.58±0.31 <![CDATA[233.34±10.32 b ]]> <![CDATA[2.21±0.18 c ]]> Experimental Group 4 Comparative Example 2 7.62±0.26 13.95±0.25 <![CDATA[211.00±11.07 c ]]> <![CDATA[2.95±0.26 b ]]> Experimental Group 5 Comparative Example 3 7.55±0.31 14.23±0.27 <![CDATA[222.65±9.97 b ]]> <![CDATA[2.97±0.23 b ]]> Experimental Group 6 Astragalus polysaccharide powder 7.62±0.27 14.20±0.32 <![CDATA[219.38±11.03 b ]]> <![CDATA[3.39±0.17 b ]]> Blank Group - 7.53±0.29 13.81±0.33 <![CDATA[209.33±12.38 c ]]> <![CDATA[6.29±0.35 a ]]>

[0101] The experimental results in Table 5 show that the average daily weight gain of Example 1, Example 2 and Comparative Example 1 was significantly increased compared with the blank group, the Comparative Example 3 and the Astragalus polysaccharide powder group were increased compared with the blank group, and there was no significant difference between Comparative Example 2 and the blank group; the diarrhea rate of each experimental group was significantly reduced compared with the blank group, among which Example 1 had the best effect.

[0102] 2.4.2 The results of serum immune index determination are shown in Table 6.

[0103] Table 6 Statistics of nonspecific immunity results of each group

[0104]

[0105] The experimental results in Table 6 show that compared with the blank group, each experimental group can significantly increase the LZM activity in piglet serum, among which Example 1 has the best effect; compared with the blank group, each experimental group can significantly increase the IgA content in piglet serum, among which Example 1 has the highest; compared with the blank group, each experimental group can significantly reduce the TNF-α content in piglet serum, among which Example 1 has the lowest; the values ​​of IgG and IgM, the results of each experimental group compared with the blank group are not significantly different.

[0106] 3. Target animal for drug efficacy evaluation experiment: Grouper:

[0107] 3.1 Experimental subjects

[0108] At a breeding farm of a biotechnology company in Haikou City, Hainan Province, groupers were domesticated in an aquarium for 3 weeks before the experiment officially began. Basic feed was fed during the domestication period. After the domestication, healthy groupers (68±5g) of the same size were selected and randomly distributed to 7 aquariums, with 30 fish in each box. The basic feed was sea fish feed; the aquarium was made of polyethylene and converted into an aerated micro-flow water system. The water flowing out of the aquarium was no longer used. The breeding water was the breeding water of the grouper breeding workshop. The breeding water was disinfected, aerated, treated with biological bags, filled with pure oxygen, and disinfected with ultraviolet rays before use.

[0109] 3.2 Experimental drugs

[0110] The experimental drugs are the drugs prepared in Examples 1 to 2 of the present invention, the drugs prepared in Comparative Examples 1 to 3, and Astragalus polysaccharide powder. The dosage of each group is: feeding twice a day (8:30-9:00, 16:30-17:00), mixed feeding, adding 400 mg per 1 kg of feed, feeding continuously for 30 days, sucking out the remaining bait with a straw 30 minutes after feeding, and keeping the box clean.

[0111] The blank group was only given the same drinking water and feed.

[0112] 3.3 Measurement indicators and measurement methods

[0113] 3.3.1 Growth indicators

[0114] Each fish was weighed at the beginning of the experiment, and the mental state and mortality of the grouper were observed during the experiment. At the end of the experiment, the fish were weighed and the weight gain rate was calculated. After the experiment, the fish were dissected.

[0115] Survival rate % = (total number of tails - number of death tails) × 100 / total number of tails

[0116] Weight gain rate % = (weight at the end of the experiment - weight at the beginning of the experiment) × 100 / weight at the beginning of the experiment

[0117] Head-kidney index % = head-kidney weight × 100 / fish weight

[0118] Spleen index % = spleen weight × 100 / fish weight

[0119] 3.3.2 Immune indicators

[0120] After the experiment, each fish was weighed after anesthesia, and the mucus on the body surface was gently scraped with tweezers to determine the LZM activity and antibacterial activity in the mucus. Blood was collected from the caudal vein with a sterile syringe and divided into two parts. One part was anticoagulated with heparin sodium to determine the phagocytic ability of leukocytes; the other part was placed in a centrifuge tube and refrigerated overnight and then centrifuged at high speed to obtain serum, and the LZM activity, NO content, albumin and total protein content in the serum were determined.

[0121] 3.4 Experimental Results

[0122] 3.4.1 Growth index results are shown in Table 7.

[0123] Table 7 Statistics of growth index results of each group

[0124] Group drug Weight gain rate (%) Head-kidney index (%) Spleen index (%) Survival rate (%) Experimental Group 1 Example 1 <![CDATA[79.56±2.87 a ]]> 0.28±0.01 0.15±0.01 100.0％ Experimental Group 2 Example 2 <![CDATA[78.12±4.31 a ]]> 0.29±0.01 0.16±0.01 100.0％ Experimental Group 3 Comparative Example 1 <![CDATA[76.69±3.71 a ]]> 0.28±0.01 0.15±0.01 100.0％ Experimental Group 4 Comparative Example 2 <![CDATA[67.12±4.35 c ]]> 0.26±0.01 0.17±0.01 100.0％ Experimental Group 5 Comparative Example 3 <![CDATA[71.36±3.61 b ]]> 0.25±0.01 0.14±0.01 100.0％ Experimental Group 6 Astragalus polysaccharide powder <![CDATA[69.15±2.98 bc ]]> 0.27±0.01 0.16±0.01 100.0％ Blank Group - <![CDATA[66.21±3.36 c ]]> 0.26±0.01 0.15±0.01 100.0％

[0125] The experimental results in Table 7 show that Example 1, Example 2 and Comparative Example 1 can significantly improve the weight gain rate of grouper, and the other groups have no significant differences compared with the blank group; the head kidney index and spleen index, compared with the blank group, the results of each experimental group have no significant differences; the survival rate of each experimental group and the blank group is 100%.

[0126] 3.4.2 Nonspecific immunity results, see Table 8.

[0127] Table 8 Statistics of nonspecific immunity results of each group

[0128]

[0129] The experimental results in Table 8 show that Example 1 and Example 2 can significantly improve the activity of LZM in the mucus on the body surface of grouper, and there is no significant difference between Comparative Examples 1 to 3 and the astragalus polysaccharide powder group and the blank group; compared with the blank group, Example 1 and Example 2 have the best antibacterial activity, Comparative Example 1 is second, and there is no significant difference between Comparative Example 3 and the blank group; compared with each group in serum LZM, Example 1> Example 2> Comparative Example 1> Astragalus polysaccharide powder group> Comparative Example 3> Comparative Example 2> Blank group, and the differences between each experimental group and the blank group are significant; compared with each group in NO level, Example 1< Example 2< Comparative Example 1< Comparative Example 3< Astragalus polysaccharide powder group< Comparative Example 2< Blank group, and the differences between each experimental group and the blank group are significant; the values ​​of albumin and A / G are not significantly different among the experimental groups.

[0130] 4. Safety experiment:

[0131] After the efficacy evaluation experiment was completed, 5 chickens, 3 pigs, and 5 fish were randomly selected from each experimental group. No obvious pathological changes were found in the internal organs and tubes, and no obvious abnormal changes were found in the tissue morphology and color. No animal deaths were found.

[0132] After the efficacy evaluation experiment was completed, the drug was stopped and the experimental animals were observed for 10 days. During the administration period and the drug withdrawal observation period, no clinical adverse reactions occurred in each experimental group; no animals in the experimental group died during the experiment. This shows that when Example 1 and Example 2 are used within the recommended dose, the clinical use of this product is safe.

[0133] This prescription has been tested for effectiveness and safety, and it has been proven that this product is safe and effective, and the effect of the embodiment is significantly better than that of the control group.

[0134] In summary, the above are only preferred embodiments of the present invention, and are not intended to limit the present invention in other forms. Any technician familiar with the profession may use the above technical content as inspiration to make changes or modifications. This is an equivalent embodiment of equivalent changes. However, any simple modification, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the technical concept of the present invention still fall within the scope of protection of the claims of the present invention.

Claims

1. A drug for improving the immunity of farmed animals, characterized in that: The active ingredients of the medicine are composed of 10 to 20 parts by weight of heptapeptide-37 of subtilisin and extracts of the following Chinese medicinal materials: 15 to 30 parts by weight of Euonymus fortunei, 15 to 30 parts by weight of Astragalus membranaceus, 15 to 30 parts by weight of Spirulina, 10 to 20 parts by weight of Silybum marianum, and 10 to 20 parts by weight of Epimedium, and the Chinese medicinal materials are calculated in terms of raw drug amount.

2. The drug according to claim 1, characterized in that The medicine comprises, by weight, 20 parts of Heptacopeptide-30 of Subtilisin, 30 parts of Euonymus fortunei, 20 parts of Astragalus, 20 parts of Spirulina, 10 parts of Milk Thistle and 10 parts of Epimedium.

3. The drug according to claim 1 or 2, characterized in that The dosage form of the drug is an oral dosage form.

4. The drug according to claim 3, characterized in that The oral dosage form is powder, granule or oral liquid.

5. The drug according to claim 1, 2 or 4, characterized in that: The extract of the Chinese medicinal material is prepared by the following steps: The Chinese medicinal materials of Euonymus fortunei, Astragalus, Spirulina, Milk thistle and Epimedium are mixed and crushed, and then an ethanol solution is added for extraction, and filtered to obtain a medicinal residue and an ethanol crude extract; Add water to the medicinal residue to extract, filter, and obtain a water extract; The ethanol in the crude ethanol extract is recovered, and the crude ethanol extract after the ethanol is recovered and the water extract are combined, and the filtrate is concentrated to 1 to 2 times the weight of the Chinese medicinal material, so as to obtain the extract of the Chinese medicinal material.

6. The drug according to claim 5, characterized in that The volume concentration of the ethanol solution is 60% to 80%, and during alcohol extraction, the total amount of Chinese medicinal materials and the ethanol solution are added in a weight ratio of 1: (8 to 12).

7. The drug according to claim 6, characterized in that The alcohol extraction is carried out by reflux, and the reflux temperature is 80°C to 90°C.

8. The drug according to claim 5, characterized in that During water extraction, the total amount of Chinese medicinal materials and water are added in a weight ratio of 1: (8-12).

9. The drug according to claim 4, 6, 7 or 8, characterized in that: The medicine is in the form of an oral liquid, which is prepared by adding heptapeptide subtilisin and a preservative to the extract of traditional Chinese medicine and mixing well.

10. The drug according to claim 4, 6, 7 or 8, characterized in that: The medicine is a powder or granule, and the extract of the Chinese medicinal material is spray-dried and then added with 77 peptide subtilisin, and then the excipient and the preservative are evenly mixed to obtain a powder; or the extract of the Chinese medicinal material is spray-dried and then added with 77 peptide subtilisin, and then the excipient, preservative, and adhesive are added, and then granulated, dried, and granulated to obtain a granule.

Images