CN113616704B - Traditional Chinese medicine preparation for treating avian influenza and preparation method thereof - Google Patents
Traditional Chinese medicine preparation for treating avian influenza and preparation method thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of veterinary drugs, and particularly relates to a traditional Chinese medicine preparation for treating avian influenza and a preparation method thereof. The traditional Chinese medicine preparation is strictly compatible according to monarch, minister, assistant and guide, the formula is optimized on the basis of the ephedra, almond, gypsum and liquorice, the amount of the roasted ephedra, the almond, the gypsum and the liquorice is reduced, the isatis root, the houttuynia cordata, the agastache, the rhubarb and the schizonepeta are added, the finally optimally prepared traditional Chinese medicine preparation has excellent inhibition effect on H9N2 subtype influenza virus, the total effective rate in clinical application is 100%, the preparation is a pure traditional Chinese medicine preparation, the formula is scientific and simple, the preparation method is simple, the dosage is small (more than 100 times of dilution) when the use concentration is reached, the cost is low, no toxicity is caused, the safety is realized, no residue is caused, the food safety can be ensured while the avian influenza is treated and prevented, and the market prospect is wide.
Description
Technical Field
The invention belongs to the technical field of veterinary drugs, and particularly relates to a traditional Chinese medicine preparation for treating avian influenza and a preparation method thereof.
Background
Influenza a virus is a highly variable pathogen that infects a variety of host species. Influenza viruses are single-stranded negative-strand RNA viruses, the genome of which consists of 8 single-stranded negative-strand RNAs, each segment encoding one or two proteins. The surface glycoproteins Hemagglutinin (HA) and Neuraminidase (NA) are the basis for the classification of influenza a viruses into subtypes. AIV HAs been classified into 18 HA subtypes and 11 NA subtypes according to antigenic differences between viral HA and NA. Avian Influenza (AIV) H9N2, which is an avian viral infectious disease caused by AIV subtype H9N2, is considered as one of the important epidemic diseases in intensive chicken farms, and is widely distributed around the world, threatening the development of the chicken industry, and often causing serious economic losses. Statistically, in 1996-2000, the chicken flock infected with H9N2 AIV accounted for 93.89%, indicating that H9N2 is the major subtype affecting poultry farming from the end of the 19 th century to the beginning of the 20 th century. To date, H9N2 is still one of the three main AI subtypes that disrupt the poultry industry, in addition to the notorious H5N1 and the emerging H7N9 newcastle. It can be said that H9N2 AIV is the most common influenza virus subtype detected in poultry populations worldwide. Recently, many poultry cases with respiratory symptoms have been clinically identified as H9N2 avian influenza and mycoplasma mixed infection by laboratory virus isolation causing chicken larynx embolism and tracheal hemorrhage.
The H9N2 subtype avian influenza vaccine alone cannot control the disease 100%, the biological safety of a farm is not in place, or improper feeding management operation can cause certain stress to chicken flocks, and the H9 outbreak can be caused. Meanwhile, because the pathogen continuously generates variation, the speed of developing the influenza vaccine cannot keep up with the variation speed of the pathogen, so that a plurality of farms immunize the vaccine and occasionally have avian influenza outbreaks. After the viral diseases occur, no specific drug therapy exists, and the mortality of animals can be obviously reduced by timely and symptomatic treatment after the viral diseases occur. Although the incidence of the disease can be reduced to a certain extent by properly adding antibiotics in the production process, the antibiotics are mainly applied to the complications caused by virus infection, and along with the wide application of a large amount of antibacterial drugs, particularly broad-spectrum antibacterial drugs in the breeding industry, the drug resistance problem of the antibacterial drugs is highlighted day by day, the public health, the sustainable growth of economy and the stability of global economy are seriously threatened by the drug resistance problem of the antibiotics, and the situation of safe drug use of livestock breeding is severe. In the guidance suggestion of promoting the health development of the veterinary drug industry, the agricultural rural area definitely needs to accelerate the development of the veterinary drug industry and ensure the safety of food and public health. By combining the unique Chinese medicine resources and the long-term history of Chinese medicines, the Chinese veterinary medicines and the byproducts thereof can be the unique resources for developing antibiotic substitutes in China.
Aiming at the defects and problems that no complete prevention and treatment measures are provided for avian influenza at present and antibiotic residues exist, a traditional Chinese veterinary medicine formula which has a remarkable inhibiting effect on H9N2 subtype influenza virus needs to be developed, and the traditional Chinese veterinary medicine formula can have good prevention and treatment effects on H9N2 subtype avian influenza clinically.
Disclosure of Invention
Aiming at the prior art, the invention aims to provide a traditional Chinese medicine preparation for treating avian influenza and a preparation method thereof. The formula of the invention is scientific and simple, and the preparation method is simple; has no toxicity, safety and residue, and can ensure food safety while treating and preventing avian influenza.
In order to achieve the purpose, the invention adopts the following technical scheme:
the invention provides a traditional Chinese medicine composition for treating avian influenza, which comprises the following traditional Chinese medicine raw materials in parts by weight:
2-4 parts of mix-fried ephedra herb, 2-4 parts of almond, 8-10 parts of gypsum, 5-7 parts of isatis root, 5-7 parts of houttuynia cordata, 2-4 parts of ageratum, 5-7 parts of rheum officinale, 2-4 parts of schizonepeta and 2-4 parts of liquorice.
Preferably, the traditional Chinese medicine composition comprises the following traditional Chinese medicine raw materials in parts by weight:
3 parts of mix-fried ephedra herb, 3 parts of almond, 9 parts of gypsum, 6 parts of isatis root, 6 parts of heartleaf houttuynia herb, 3 parts of ageratum, 6 parts of rhubarb, 3 parts of fineleaf schizonepeta herb and 3 parts of liquorice.
In a second aspect of the present invention, a preparation method of the above traditional Chinese medicine composition is provided, wherein the method comprises:
(1) mixing the Chinese medicinal materials, adding 25 deg.C water, soaking for 30 min, heating to boil and maintaining for 1 hr, filtering to obtain first extractive solution, adding 50 deg.C water to the filtered Chinese medicinal materials, heating to boil and maintaining for 45 min, and filtering to obtain second extractive solution;
(2) mixing the first extractive solution and the second extractive solution to obtain the Chinese medicinal composition.
Preferably, in the step (1), the adding amount of the traditional Chinese medicine raw materials and water with the temperature of 25 ℃ is 1g (10-15) mL.
Preferably, in the step (2), the adding amount of the traditional Chinese medicine raw materials and 50 ℃ water is 1g (5-7) mL.
In a third aspect of the invention, an application of a traditional Chinese medicine composition in preparing a traditional Chinese medicine preparation for treating avian influenza is provided.
Preferably, the avian influenza is influenza subtype H9N 2.
In a fourth aspect of the invention, a traditional Chinese medicine preparation for treating avian influenza is provided, which takes a traditional Chinese medicine composition as an effective component.
Preferably, the Chinese medicinal preparation further comprises pharmaceutically acceptable auxiliary materials.
The invention has the beneficial effects that:
the traditional Chinese medicine preparation disclosed by the invention is strictly compatible according to monarch, minister, assistant and guide, and the traditional Chinese medicine preparation prepared by optimizing the formula on the basis of the ephedrin has an excellent inhibition effect on H9N2 subtype influenza viruses, has a total effective rate of 100% in clinical application, is a pure traditional Chinese medicine preparation, is scientific and simple in formula, simple in preparation method, small in dosage (more than 100 times of dilution) in use concentration, low in cost, free of any toxicity, safe and free of residue, can ensure food safety while treating and preventing avian influenza, and has a wide market prospect.
Drawings
FIG. 1: comparing the development conditions of the chick embryos, wherein A. the traditional Chinese medicine stock solution prepared according to the example 1, B. the traditional Chinese medicine stock solution is diluted by 10 times, C. the traditional Chinese medicine stock solution is diluted by 100 times, and D. a normal control;
FIG. 2 is a schematic diagram: comparison of chick embryo development, wherein A, B is diluted 10 times in the traditional Chinese medicine prepared according to the method of example 1; C. d is 100 times diluted of the traditional Chinese medicine prepared according to the method of the embodiment 1; MXSG is a diluted group of 10 times of the traditional Maxingshigan water extract. 1 represents: traditional Chinese medicine + virus, 2 represents: viral control, 3 represents: and (5) traditional Chinese medicine control.
Detailed Description
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
As introduced in the background section, the problems of incomplete prevention and treatment measures for avian influenza, defects of antibiotic residues and the like exist at present.
Based on the above, the invention aims to provide the traditional Chinese medicine preparation for treating avian influenza and the preparation method thereof, and the traditional Chinese medicine preparation has the advantages of small dosage, low cost, no toxicity, safety and no residue. The formula is optimized on the basis of the ephedra and apricot stone glycoside, the consumption of the ephedra and apricot stone glycoside is reduced by about half, and the isatis root, the houttuynia cordata, the ageratum, the rhubarb and the fineleaf schizonepeta herb are added; the total dosage of the raw material medicines is basically equivalent to that of the ephedra, almond and gypsum in the traditional formula, but the effect of treating H9N2 subtype influenza is far better than that of the ephedra, almond and gypsum.
In order to make the technical solutions of the present application more clearly understood by those skilled in the art, the technical solutions of the present application will be described in detail below with reference to specific embodiments.
The test materials used in the examples of the present invention are all conventional in the art and commercially available.
Example 1
Weighing 3g of mix-fried ephedra herb, 3g of almond, 9g of gypsum, 6g of isatis root, 6g of heartleaf houttuynia herb, 3g of ageratum, 6g of rhubarb, 3g of fineleaf schizonepeta herb and 3g of liquorice, uniformly mixing, putting into a container, adding 500ml of clean water at 25 ℃, soaking for 30 minutes, heating to boil and keeping for 1 hour, collecting an extracting solution, adding 250ml of clean water at 50 ℃, heating to boil and keeping for 45 minutes, and collecting the extracting solution; mixing the two extractive solutions, and filtering to obtain compound Chinese medicinal water extractive solution.
Example 2
Weighing 2g of mix-fried ephedra herb, 4g of almond, 8g of gypsum, 7g of isatis root, 5g of heartleaf houttuynia herb, 4g of ageratum, 5g of rhubarb, 4g of fineleaf schizonepeta herb and 2g of liquorice, uniformly mixing, putting into a container, adding 500ml of clean water at 25 ℃, soaking for 30 minutes, heating to boil and keeping for 1 hour, collecting an extracting solution, adding 250ml of clean water at 50 ℃, heating to boil and keeping for 45 minutes, and collecting the extracting solution; mixing the two extractive solutions, and filtering to obtain compound Chinese medicinal water extractive solution.
Example 3
Weighing 4g of mix-fried ephedra herb, 2g of almond, 10g of gypsum, 5g of isatis root, 7g of heartleaf houttuynia herb, 2g of ageratum, 7g of rhubarb, 2g of fineleaf schizonepeta herb and 4g of liquorice. Mixing, placing into a container, adding 500ml of 25 deg.C clear water, soaking for 30 min, heating to boil and maintaining for 1 hr, collecting extractive solution, adding 250ml of 50 deg.C clear water again, heating to boil and maintaining for 45 min, and collecting extractive solution; mixing the two extractive solutions, and filtering to obtain compound Chinese medicinal water extractive solution.
Comparative example
Preparing traditional Maxingshigan water extract
Weighing 6g of mix-fried ephedra, 6g of almond, 18g of gypsum and 6g of liquorice, uniformly mixing, putting into a container, adding 500ml of 25 ℃ clear water, soaking for 30 minutes, heating to boil and keeping for 1 hour, collecting an extracting solution, adding 250ml of 50 ℃ clear water again, heating to boil and keeping for 45 minutes, and collecting the extracting solution; mixing the two extractive solutions, and filtering to obtain compound Chinese medicinal water extractive solution.
Test example 1: drug safety evaluation
Experimental materials:
chicken embryo: 10-day-old SPF chick embryos, supplied by the SPF center of the poultry institute of agronomy, Shandong province.
The experimental method comprises the following steps: the preparation method of example 1 is used to prepare a compound traditional Chinese medicine water extract, the extract is divided into three parts averagely, one part is recorded as a stock solution without treatment, the second part is recorded as 10 times diluted by 10 times, the third part is recorded as 100 times diluted by 100 times, SPF (specific pathogen free) chick embryos of 10 days old are inoculated according to the dosage of 100 ul/embryo, chick embryos which die within 24 hours are removed, the death condition of the chick embryos is recorded, and the development condition of the chick embryos is observed after 5 days.
The experimental results are as follows: as shown in figure 1, compared with the blank control group of chick embryos, the compound traditional Chinese medicine water extract prepared in example 1 and diluted by 10 times and 100 times are not used for influencing the development of the chick embryos, and the chick embryos do not die.
Therefore, the traditional Chinese medicine compound provided by the invention has good safety and is nontoxic to chick embryos.
Test example 2: evaluation of anti-influenza Virus Effect of drugs
Experimental Material
Chicken embryo: 10-day-old SPF chick embryos provided by SPF center, poultry institute of agricultural sciences, Shandong province;
virus strain: epidemic strains of influenza subtype H9N2 were isolated, identified, purified and amplified by the animal husbandry and veterinary institute of academy of agricultural sciences, Shandong province.
Experimental methods
Half the infection amount (EID) of the virus 50 ) And (3) determination: serially diluting H9N2 influenza virus 10 times, inoculating SPF chick embryo of 10 days old, inoculating embryo (0.1 ml/embryo) at each dilution of 5 pieces, and removing within 24HThe allantoic fluid of the dead chick embryos was collected after 5 days to determine the virus titer, and half of the infectious dose of the virus was calculated.
Evaluation of antiviral effect of drug: according to measured EID 50 Establishing an H9N2 influenza virus infected chick embryo model, evaluating the antiviral effect, diluting the prepared medicament by 10 times and 100 times respectively according to the method of the embodiment 1, mixing and incubating the medicament with the influenza virus in equal amount, and inoculating 10-day SPF chick embryos according to the dose of 200 ul/embryo. The traditional ephedra, almond and licorice medicine prepared in the comparative example is diluted by 10 times, mixed and incubated with influenza virus in equal amount, and SPF chick embryos of 10 days old are inoculated according to the dosage of 200 ul/embryo. Removing dead chick embryos within 24h, recording the death condition of the chick embryos, collecting allantoic fluid after 5 days to measure the virus titer, and observing the development condition of the chick embryos.
The experimental results are as follows: as shown in table 1 and fig. 2, the prepared herbal compound was diluted 10-fold and 100-fold according to the method of example 1, and thus the replication of influenza virus in chick embryos could be effectively controlled. The effect of the traditional Chinese medicine compound prepared in the example 1 after being diluted by 100 times is slightly better than the effect of the traditional classic formula of the comparative example, namely the gypsum and licorice, after being diluted by 10 times, the effect of the traditional Chinese medicine compound prepared in the example 1 is obviously better than the effect of the traditional classic formula of the comparative example, namely the gypsum and licorice, after being diluted by 10 times. The formula of the invention has obvious effect on treating H9N2 influenza.
Therefore, the traditional Chinese medicine compound provided by the invention has a good antiviral effect.
TABLE 1
| MXSG (10 times) | MXSG (10 times) | A (10 times) | B (10 times) | C (100 times) | D (100 times) | |
| Medicine + toxin | 9/0/0 | 10/0/0 | 0/0/0 | 0/0/0 | 9/0/0 | 0/0/0 |
| Toxicity control | 11/11/11 | 10/10/9 | 11/11/11 | 10/10/9 | 11/10/10 | 10/9/9 |
| Drug control | 0/0/0 | 0/0/0 | 0/0/0 | 0/0/0 | 0/0/0 | 0/0/0 |
And (4) surface note: the numbers in the table represent the hemagglutination titer of chick embryo allantoic fluid (i.e., the virus content in chick embryos), the bold font indicates that the chick embryo died within 5 days, and the bold font indicates that the chick embryo survived within 5 days.
Test example 3: clinical application effects (treatment examples)
The application effect of the farm A is as follows: a place: a jujube garden.
26 thousands of big meat food, 24 days old, 60% of time control of a fan, overlarge ventilation volume, quick reduction of the external temperature in the same day, no reduction of the fan and a small window in time, cold of chickens, lengthening of eyes, hair explosion, respiratory symptoms and no material increase, typical disease materials are collected by sick chicken flocks, and the bird flu is positive by laboratory detection of H9N 2.
The traditional Chinese medicine compound preparation is prepared according to the method of example 1 in treatment, 8 jin of water is mixed in each ml of the traditional Chinese medicine compound preparation, after the traditional Chinese medicine compound preparation is continuously used for 4 days, the respiratory tract of a large group is not developed and is slowly reduced, the death and culling of chicken groups are not increased, the feeding is recovered, and finally the marketing result is as follows: the weight is 5.77, the survival rate is 96 percent, the feed ratio is 1.58 (the feed ratio is higher when new corns are used), chickens emerge in 43 days, and the weight ratio of Europe and the European finger: 408. the next chicken farm also has the same symptoms, and the treatment according to other schemes is uncontrollable and extremely high in death and culling.
Test example 4: clinical effects (prevention example)
The application effect of the farm B is as follows: a place: in the south of the river.
The next two breeders are neighbors, and the breeder varieties are 817 broiler chickens, which are bred in cages at 40-day age, one is 1.6 thousands, and the other is 1.8 thousands. 1.6 ten thousand farmers use the Chinese herbal compound preparation prepared according to the method of the embodiment 1, mix 8 jin of water per ml of the Chinese herbal compound preparation, and continuously use the mixture for three days; 1.8 ten thousand farmers did not have the herbal compound preparation prepared according to the method of example 1; the chicken is blown suddenly for cooling at the age of 43 days, 1.6 thousands of traditional Chinese medicines are used, the disease is not caused, and the chicken flocks are normal; 1.8 thousands of Chinese medicaments are not used, respiratory symptoms are caused, and materials are reduced at the same time, so that the Chinese medicinal compound preparation has the effect of preventing cold.
The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims (8)
1. A traditional Chinese medicine composition for treating H9N2 subtype avian influenza is characterized by being prepared from the following traditional Chinese medicine raw materials in parts by weight:
2-4 parts of mix-fried ephedra herb, 2-4 parts of almond, 8-10 parts of gypsum, 5-7 parts of isatis root, 5-7 parts of houttuynia cordata, 2-4 parts of ageratum, 5-7 parts of rheum officinale, 2-4 parts of schizonepeta and 2-4 parts of liquorice.
2. The traditional Chinese medicine composition according to claim 1, which is prepared from the following traditional Chinese medicine raw materials in parts by weight:
3 parts of mix-fried ephedra herb, 3 parts of almond, 9 parts of gypsum, 6 parts of isatis root, 6 parts of heartleaf houttuynia herb, 3 parts of ageratum, 6 parts of rhubarb, 3 parts of fineleaf schizonepeta herb and 3 parts of liquorice.
3. A method for preparing the traditional Chinese medicine composition of claim 1 or 2, which is characterized in that the method comprises the following steps:
(1) mixing the Chinese medicinal materials, adding 25 deg.C water, soaking for 30 min, heating to boil and maintaining for 1 hr, filtering to obtain first extractive solution, adding 50 deg.C water to the filtered Chinese medicinal materials, heating to boil and maintaining for 45 min, and filtering to obtain second extractive solution;
(2) mixing the first extractive solution and the second extractive solution to obtain the Chinese medicinal composition.
4. The preparation method according to claim 3, wherein in the step (1), the Chinese medicinal raw materials are uniformly mixed, and when water at 25 ℃ is added, the adding amount ratio of the Chinese medicinal raw materials to the water at 25 ℃ is 1g: 10-15 mL.
5. The preparation method according to claim 3, wherein in the step (1), when water of 50 ℃ is added to the filtered traditional Chinese medicine raw materials, the ratio of the added amount of the traditional Chinese medicine raw materials to the water of 50 ℃ is 1g: 5-7 mL.
6. The use of the Chinese medicinal composition of claim 1 or 2 in the preparation of a Chinese medicinal preparation for treating H9N2 subtype avian influenza.
7. A traditional Chinese medicine preparation for treating H9N2 subtype avian influenza, which is characterized in that the traditional Chinese medicine composition of claim 1 or 2 is used as an effective component.
8. The traditional Chinese medicine preparation according to claim 7, characterized in that: the traditional Chinese medicine preparation also comprises pharmaceutically acceptable auxiliary materials.
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