A traditional Chinese medicine composition for preventing and treating bee paralysis disease, its preparation method and application

Abstract

The present invention discloses a traditional Chinese medicine composition for preventing and treating bee paralysis disease, its preparation method and application, belonging to the field of traditional Chinese medicine preparations. The traditional Chinese medicine composition, by weight, comprises the following components: 5-10 parts of Scutellaria baicalensis, 5-10 parts of Ginkgo biloba leaves, 3-8 parts of Pueraria lobata, 3-8 parts of Glycyrrhiza uralensis, 5-8 parts of Hemerocallis fulva roots, 5-8 parts of Aconitum carmichaelii, 1-5 parts of Taraxacum mongolicum, 1-5 parts of Crataegus pinnatifida and 1-3 parts of Rheum officinale. The traditional Chinese medicine composition has the effects of clearing heat and detoxifying, promoting metabolism and enhancing immunity, has outstanding prevention and treatment effects on bee paralysis disease, can increase the survival rate of infected bee colonies to more than 75%, and has safe components and small side effects.

Description

Technical Field

[0001] The present invention relates to the field of traditional Chinese medicine preparations, and particularly to a traditional Chinese medicine composition for preventing and treating bee paralysis disease, its preparation method and application. Background Art

[0002] Bees are important economic insects. In agricultural production, bee pollination can significantly improve the quality and yield of agricultural products, and at the same time can provide natural and high-quality bee products. However, bees are vulnerable to various pathogens, such as viruses, bacteria and parasites. Among them, bee infectious diseases have become an important factor affecting the yield and safety of bee products due to their serious harm, rapid spread and easy recurrence of the disease. Viral diseases refer to infectious diseases caused by bee viruses. Currently, more than 20 kinds of bee viral diseases are known. Among them, bee paralysis disease is an infectious disease that harms adult bees caused by bee paralysis virus, which is divided into acute paralysis disease and chronic paralysis disease, and is caused by Acute Bee Paralysis Virus (ABPV) and Chronic Bee Paralysis Virus (CBPV) respectively. In a bee colony, bee paralysis disease is very common and difficult to cure once infected. When a large-scale outbreak of bee paralysis disease occurs, the development of the bee colony and the yield of bee products per single colony will be seriously affected. At the same time, it will also affect the normal pollination of bees, causing huge economic losses to agriculture and the bee production and breeding industry.

[0003] The biggest characteristic of a virus is that it mutates very quickly and is easy to escape the resistance of the host or drugs, thus changing the sequence of the original gene, making it impossible to effectively control viral diseases, and there is a risk of contaminating bee products. In addition, due to the short lifespan of bees, it is very difficult or impossible to cure infected bees with drugs. Therefore, the drugs used to prevent and treat bee paralysis disease are to prophylactically control the bees that have not been infected with the pathogen in the diseased bee colony and other bee colonies in the farm that have not been infected with the pathogen, so that they will no longer be infected and get sick. However, the current drugs for preventing and treating bee paralysis disease generally have problems such as large side effects or low obvious efficiency. Therefore, a drug that can effectively prevent and treat bee paralysis is needed. Summary of the Invention

[0004] The purpose of the present invention is to provide a traditional Chinese medicine composition for preventing and treating bee paralysis disease, its preparation method and application, so as to solve the problems existing in the above-mentioned prior art. This traditional Chinese medicine composition has the effects of clearing heat and detoxifying, promoting metabolism and enhancing immunity, has outstanding prevention and treatment effects on bee paralysis disease, and has safe ingredients and small side effects.

[0005] To achieve the above purpose, the present invention provides the following solutions:

[0006] The present invention provides a traditional Chinese medicine composition for preventing and treating bee paralysis disease, which comprises the following components by weight: 5-10 parts of Scutellaria baicalensis, 5-10 parts of Ginkgo biloba leaves, 3-8 parts of Pueraria lobata, 3-8 parts of Glycyrrhiza uralensis, 5-8 parts of Hemerocallis fulva roots, 5-8 parts of Aconitum carmichaelii, 1-5 parts of Taraxacum mongolicum, 1-5 parts of Crataegus pinnatifida and 1-3 parts of Rheum officinale.

[0007] Further, it comprises the following components by weight: 10 parts of Scutellaria baicalensis, 10 parts of Ginkgo biloba leaves, 5 parts of Pueraria lobata, 5 parts of Glycyrrhiza uralensis, 8 parts of Hemerocallis fulva roots, 5 parts of Aconitum carmichaelii, 3 parts of Taraxacum mongolicum, 3 parts of Crataegus pinnatifida and 1 part of Rheum officinale.

[0008] The present invention also provides an application of the above-mentioned traditional Chinese medicine composition in the preparation of a drug for preventing and treating bee paralysis disease.

[0009] The present invention also provides a drug for preventing and treating bee paralysis disease, the raw material of which comprises the above-mentioned traditional Chinese medicine composition.

[0010] Further, the drug also comprises a pharmaceutically acceptable excipient.

[0011] Further, the dosage form of the drug includes a spray, a powder, a granule or an ointment.

[0012] The present invention also provides a preparation method of the above-mentioned drug, which comprises the steps of extracting the active ingredients from the above-mentioned traditional Chinese medicine composition by using a solvent and then preparing the drug.

[0013] Further, the extraction method of the active ingredients includes: after crushing the traditional Chinese medicine composition, adding the solvent for preliminary extraction, filtering to obtain a first extract and residues, adding the residues to the solvent for reflux extraction to obtain a second extract, combining the first extract and the second extract, recovering and concentrating to obtain the active ingredients.

[0014] Further, during the preliminary extraction, the mass ratio of the material to the liquid is 1:3; during the reflux extraction, the mass ratio of the material to the liquid is 1:2.

[0015] Further, the solvent is an ethanol solution with a volume percentage concentration of 60%.

[0016] The present invention discloses the following technical effects:

[0017] In the traditional Chinese medicine composition of the present invention, Scutellaria baicalensis, Ginkgo biloba leaves and Hemerocallis fulva roots have the effect of clearing heat and detoxifying. The extract contains a large amount of flavonoid compounds, which have anti-inflammatory properties, can inhibit the inflammatory response, reduce the release of inflammatory mediators, and regulate the immune system response, thereby enhancing immunity; Hawthorn and Licorice can strengthen the stomach and promote digestion, and are combined with Rhubarb to guide the medicine downward, which helps to promote the excretion of bees. This traditional Chinese medicine composition has the effects of clearing heat and detoxifying, promoting metabolism and enhancing immunity, has outstanding prevention and treatment effects on bee paralysis disease, and has safe ingredients and little side effects. Detailed implementation manners

[0018] The various exemplary implementation manners of the present invention will now be 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 implementation manners of the present invention.

[0019] It should be understood that the terms used in the present invention are only for describing specific implementation manners and are not used 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. Any intermediate value within any stated value or stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded from the range.

[0020] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention pertains. Although the present invention only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein can also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the documents. In case of conflict with any incorporated document, the content of this specification shall prevail.

[0021] Without departing from the scope or spirit of the present invention, various improvements and changes can be made to the specific implementation manners of the present invention specification, which are obvious to those skilled in the art. Other implementation manners obtained from the present invention specification are obvious to those skilled in the art. The present invention specification and examples are only exemplary.

[0022] Regarding the terms "comprising", "including", "having", "containing", etc. used herein, they are all open-ended terms, that is, they are meant to include but not limited to.

[0023] Example 1

[0024] (1) Preparation of medicine

[0025] Weigh 10 parts of Scutellaria baicalensis, 10 parts of Ginkgo biloba leaves, 5 parts of Pueraria lobata, 5 parts of Glycyrrhiza uralensis, 8 parts of Hemerocallis fulva roots, 5 parts of Aconitum carmichaelii, 3 parts of Taraxacum mongolicum, 3 parts of Crataegus pinnatifida, and 1 part of Rheum palmatum by weight. Select clean materials, wash them, dry them, and set aside.

[0026] (2) Extraction

[0027] After crushing the raw materials dried in step (1), add an ethanol solution with a volume percentage concentration of 60% according to the mass ratio of material to liquid of 1:3, soak for 1 h, filter to obtain the first extract and the filter residue.

[0028] Add the obtained filter residue to an ethanol solution with a volume percentage concentration of 60% according to the mass ratio of material to liquid of 1:2, and reflux for 10 extractions to obtain the second extract.

[0029] (3) Recovery and concentration

[0030] Combine the first extract and the second extract in step (2), place them in a rotary evaporator for concentration, and concentrate at 55 °C to a relative density of 1.2 to obtain the traditional Chinese medicine extract.

[0031] Example 2

[0032] (1) Medicine preparation

[0033] Weigh 8 parts of Scutellaria baicalensis, 5 parts of Ginkgo biloba leaves, 3 parts of Pueraria lobata, 8 parts of Glycyrrhiza uralensis, 5 parts of Hemerocallis fulva roots, 8 parts of Aconitum carmichaelii, 5 parts of Taraxacum mongolicum, 1 part of Crataegus pinnatifida, and 3 parts of Rheum palmatum by weight. Select clean materials, wash them, dry them, and set aside.

[0034] (2) Extraction

[0035] After crushing the raw materials dried in step (1), add an ethanol solution with a volume percentage concentration of 60% according to the mass ratio of material to liquid of 1:3, soak for 1.5 h, filter to obtain the first extract and the filter residue.

[0036] Add the obtained filter residue to an ethanol solution with a volume percentage concentration of 60% according to the mass ratio of material to liquid of 1:2, and reflux for 20 extractions to obtain the second extract.

[0037] (3) Recovery and concentration

[0038] Combine the first extract and the second extract in step (2), place them in a rotary evaporator for concentration, and concentrate at 55 °C to a relative density of 1.3 to obtain the traditional Chinese medicine extract.

[0039] Example 3

[0040] (1) Medicine preparation

[0041] Weigh 5 parts of Scutellaria baicalensis, 8 parts of Ginkgo biloba leaves, 8 parts of Pueraria lobata, 3 parts of Glycyrrhiza uralensis, 7 parts of Hemerocallis fulva roots, 7 parts of Aconitum carmichaelii, 1 part of Taraxacum mongolicum, 5 parts of Crataegus pinnatifida and 2 parts of Rheum palmatum by weight. Select clean materials, wash them, dry them and set aside.

[0042] (2) Extraction

[0043] After crushing the raw materials dried in step (1), add an ethanol solution with a volume percentage concentration of 60% according to the material-liquid mass ratio of 1:3, soak for 2 h, filter to obtain the first extract and the filter residue.

[0044] Add the obtained filter residue to an ethanol solution with a volume percentage concentration of 60% according to the material-liquid mass ratio of 1:2, and reflux for 15 extractions to obtain the second extract.

[0045] (3) Recovery and concentration

[0046] Combine the first extract and the second extract in step (2), place them in a rotary evaporator for concentration, and concentrate to a relative density of 1.2 at 55 °C to obtain a traditional Chinese medicine extract.

[0047] Comparative example 1

[0048] Same as Example 1, the only difference is that the Hemerocallis fulva roots are removed from the raw materials.

[0049] Comparative example 2

[0050] Same as Example 1, the only difference is that the Ginkgo biloba leaves are replaced with Osmanthus fragrans leaves.

[0051] Comparative example 3

[0052] Same as Example 1, the only difference is that the Scutellaria baicalensis is replaced with Coptis chinensis.

[0053] Comparative example 4

[0054] Same as Example 1, the only difference is that the Pueraria lobata is replaced with Bupleurum chinense.

[0055] Effect verification

[0056] I. Safety test

[0057] (1) Add the traditional Chinese medicine extracts prepared in Examples 1-3 and Comparative examples 1-4 to 50% sucrose solution at final concentrations of 0.1 μg / mL, 10 μg / mL, 50 μg / mL, 100 μg / mL and 500 μg / mL, and feed healthy bees. There are 30 bees in each group, and there are 3 groups for each dose. Feed continuously for 10 days.

[0058] (2) In addition, there are two groups. One group is fed only with PBS buffer solution as a negative control, and the other group is fed only with the above 50% sucrose solution as a blank control. Record the number of dead individuals and the mortality rate of each group of bees every day for all groups.

[0059] The results showed that no individual death was found in any group, and this traditional Chinese medicine extract would not affect the health of the bee colony.

[0060] II. Prevention and control effect test

[0061] In August 2021, 100 boxes of bees from 3 apiaries were selected in Yanji City, Jilin Province for the experiment. They were randomly divided into 10 groups, with 10 boxes of bee colonies in each group, and each box of bee colonies had 9000 - 10000 bees. Among them, 7 groups were experimental groups, and the traditional Chinese medicine extracts prepared in Examples 1 - 3 and Comparative Examples 1 - 4 were added to the 50% sucrose solution containing bee paralysis virus at a dosage of 100 μg / mL to feed the bees in the experimental groups. The other 3 groups were control groups. One group was fed only with PBS buffer solution as a negative control (PBS), another group was fed only with the 50% sucrose solution containing the virus as the virus group (PV), and the third group was fed only with the 50% sucrose solution without the virus as a blank control (ck). The viruses in the above experimental groups and blank groups were obtained by ultracentrifugation and added to the 50% sucrose solution at a copy number of about 10 12 copies per microliter for feeding. The number of dead bees in each group was recorded every 3 days, the survival rate was calculated, the dead bees were cleaned before the beehive was cleaned in the early morning, and the number of bees showing paralysis symptoms was recorded, and the infection rate was calculated. The results are shown in Tables 1 - 2.

[0062] Table 1 Survival rate of bees in each group (%)

[0063]

[0064] Table 2 Percentage of the number of bees with paralysis symptoms (%)

[0065]

[0066] As can be seen from Tables 1 - 2, compared with the control groups, the survival rates of the bees in the experimental groups were generally increased. Feeding the traditional Chinese medicine extracts prepared in Examples 1 - 4 could significantly improve the survival rate of the bee colonies infected with bee paralysis disease. The survival rate of the bee colonies remained above 75% after 15 days; the number of bees with paralysis symptoms in the bee colonies decreased significantly, indicating that this traditional Chinese medicine extract could enhance the disease resistance of bees to bee paralysis virus.

[0067] The above - described embodiments are only descriptions of the preferred embodiments of the present invention, and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, various deformations and improvements made by those of ordinary skill in the art to the technical solutions of the present invention shall fall within the protection scope determined by the claims of the present invention.

Claims

1. A Chinese medicine composition for preventing and treating bee paralysis, characterized in that: The invention is composed of the following components by weight: 10 parts of scutellaria baicalensis, 10 parts of ginkgo leaves, 5 parts of kudzu root, 5 parts of liquorice, 8 parts of daylily root, 5 parts of aconite, 3 parts of dandelion, 3 parts of hawthorn and 1 part of rhubarb.

2. Use of the Chinese medicine composition as claimed in claim 1 in the preparation of medicines for preventing and treating bee paralysis.

3. A drug for preventing and treating bee paralysis, characterized in that: The raw materials consist of the traditional Chinese medicine composition according to claim 1 and pharmaceutically acceptable excipients.

4. The drug according to claim 3, characterized in that The dosage form of the drug includes spray, powder, granule or ointment.

5. A method for preparing the drug according to claim 3 or 4, characterized in that: The method comprises the steps of obtaining the effective ingredients in the traditional Chinese medicine composition according to claim 1 by solvent extraction, and then preparing the medicine.

6. The preparation method according to claim 5, characterized in that: The method for extracting the active ingredient comprises: crushing the traditional Chinese medicine composition, adding the solvent for preliminary extraction, filtering to obtain a first extract and a filter residue, adding the solvent to the filter residue for reflux extraction to obtain a second extract, combining the first extract and the second extract, recovering and concentrating to obtain the active ingredient.

7. The preparation method according to claim 6, characterized in that: During the initial extraction, the mass ratio of the feed to the liquid is 1:3; during the reflux extraction, the mass ratio of the feed to the liquid is 1:

2.

8. The preparation method according to claim 6, characterized in that: The solvent is an ethanol solution with a volume percentage concentration of 60%.

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