A pharmaceutical composition for expelling fish parasites and its preparation method and application

Abstract

This invention provides a pharmaceutical composition for expelling fish parasites, its preparation method, and its application, belonging to the field of aquaculture technology. The pharmaceutical composition comprises: neem bark, Dryopteris crassirhizoma, Artemisia argyi, Artemisia annua, and Moringa root. The composition is scientifically formulated based on the anthelmintic properties, synergistic effects, and penetrating power of each ingredient, achieving a broad-spectrum, targeted, and highly effective anthelmintic effect. This composition is highly effective against various parasites such as Gyrodactylus, Trichodina, Chilodonella, Bodoworm, and Gyrodactylus, broadening the anthelmintic spectrum and enhancing its ability to kill parasites in the gills, resulting in thorough expulsion. By reducing drug concentration and adding auxiliary components, the composition improves drug utilization and reduces stress responses in fish, making it more suitable for green aquaculture compared to the high-concentration methods used in existing technologies.

Description

Technical Field

[0001] This invention belongs to the field of aquaculture technology, specifically relating to a pharmaceutical composition for expelling fish parasites, its preparation method, and its application. Background Technology

[0002] In aquaculture, parasites such as Gyrodactylus, Trichodina, Chilodonella, Bodoworm, and Caucasian are common sources of disease that harm fish health. In particular, these parasites tend to infest the gills, which are important respiratory organs for fish. Once the gills are invaded by parasites, it can lead to difficulty breathing, loss of appetite, and slow growth. In severe cases, it can cause large-scale mortality and result in huge economic losses for fish farmers. Currently, some drugs exist for treating fish parasites. While these drugs are effective against some parasites, they have significant limitations: First, their spectrum of action is narrow, making it difficult to simultaneously kill multiple parasites such as Gyrodactylus, Trichodina, Chilodonella, Bottomella, and Gyrodactylus. Farmers often need to use multiple drugs in combination, which is cumbersome. Second, for parasites residing in the gills, existing drugs lack sufficient penetration and targeting, making it difficult to effectively target parasites deep within the gills, resulting in incomplete treatment and recurring symptoms. Third, some drugs require high concentrations to achieve a certain effect, which may cause stress in fish and affect their normal growth. Furthermore, while some existing technologies involve simple combinations of herbal medicines, the lack of scientific ratio optimization and failure to fully utilize the synergistic effects between different drug components leads to poor overall treatment efficacy, failing to meet farmers' needs for highly efficient, broad-spectrum, and targeted treatment of fish parasites, especially those residing in the gills. Summary of the Invention

[0003] In view of the deficiencies in the prior art, the purpose of this invention is to provide a pharmaceutical composition that can achieve broad-spectrum killing of various fish parasites such as Gyrodactylus, Trichodina, Chilodonella, Bodoworm, and Cupella, and effectively kill parasites in the gills of fish.

[0004] The objective of this invention is achieved through the following technical solution: This invention provides a pharmaceutical composition comprising, by weight: 1.5-2 parts of neem bark, 1-1.5 parts of Dryopteris crassirhizoma, 1-1.5 parts of Artemisia argyi, 0.8-1.2 parts of Artemisia annua, and 0.5-0.8 parts of Moringa root.

[0005] Preferably, the pharmaceutical composition comprises, by weight parts: 1.8 parts of neem bark, 1.2 parts of Dryopteris crassirhizoma, 1.2 parts of Artemisia argyi, 1.0 part of Artemisia annua, and 0.6 parts of Moringa root.

[0006] Preferably, the pharmaceutical composition further includes Tween-80 and ethanol.

[0007] This invention provides a method for preparing the pharmaceutical composition described above, comprising the following steps: The bark of neem tree, dryopteris crassirhizoma, mugwort leaves, artemisia annua and moringa root are crushed and mixed to obtain a raw material mixture; The raw material mixture is boiled to obtain an aqueous extract; The aqueous extract was concentrated to obtain a concentrated solution; The concentrate was mixed with ethanol and Tween-80 to obtain a pharmaceutical composition.

[0008] Preferably, the method for decocting the raw material mixture includes: Mix the raw material mixture with water and boil it for the first time. Collect the first decoction and the residue separately. Mix the filter residue with water and boil it a second time, then collect the second decoction. The first decoction and the second decoction are mixed to obtain an aqueous extract.

[0009] Preferably, the mass ratio of the raw material mixture to water is 1:(10~15); the mass ratio of the filter residue to water is 1:(8~10).

[0010] Preferably, the first decoction time is 1.5 to 2 hours; the second decoction time is 1 hour.

[0011] The present invention provides the application of the pharmaceutical composition described in the above technical solution or the pharmaceutical composition prepared by the preparation method described in the above technical solution in the treatment of fish parasites.

[0012] Preferably, the fish species include any one or more of carp, grass carp, and crucian carp; the parasites include any one or more of Dactylogyrus, Trichodina, Chilodonella, Bodo, and Cercozoa.

[0013] The present invention provides a method for expelling parasites, comprising: applying the drug composition described in the above technical solution or the drug composition prepared by the preparation method described in the above technical solution to the water body for aquaculture fish.

[0014] The beneficial effects of this invention are: This invention provides a pharmaceutical composition comprising, by weight: 1.5-2 parts of neem bark, 1-1.5 parts of Dryopteris crassirhizoma, 1-1.5 parts of Artemisia argyi, 0.8-1.2 parts of Artemisia annua, and 0.5-0.8 parts of Moringa root. The pharmaceutical composition provided by this invention is not a simple superposition of existing herbal medicines, but rather a scientifically formulated combination based on the insecticidal properties, synergistic effects, and penetrating power of each medicinal component. For the first time, neem bark, Dryopteris crassirhizoma, Artemisia argyi, Artemisia annua, and Moringa root are combined with specific auxiliary ingredients to achieve a broad-spectrum, targeted, and highly effective insecticidal synergistic effect, solving the problem of unscientific formulation in existing technologies. The drug composition described herein offers a more comprehensive anthelmintic effect: Existing technologies often rely on single drugs or simple combinations to simultaneously target multiple parasites such as Gyrodactylus, Trichodina, Chilodonella, Bodina, and Gyrodina, and are less effective against deep-seated parasites in the gills. The drug composition of this invention, through component optimization and ratio adjustment, efficiently kills multiple parasites including Gyrodactylus, Trichodina, Chilodonella, Bodina, and Gyrodina, broadening the anthelmintic spectrum and enhancing its ability to kill parasites in the gills. Its thoroughness in anthelminsis is significantly superior to existing technologies. Furthermore, while ensuring anthelmintic efficacy, the dosage of each component in the drug composition is no higher (or lower) than the concentration of a single anthelmintic. The synergistic effect produced by this scientific compounding significantly improves drug utilization and reduces stress on fish. Compared to the high-concentration application of single drugs in existing technologies, this approach better meets the needs of green aquaculture. Moreover, the drug composition provided by this invention is more practical. The drug composition is simple to prepare and easy to use, requiring no professional equipment or complicated operations. It is suitable for promotion and application by various aquaculture farmers, solving the problems of cumbersome operation and difficulty in promotion of some existing deworming solutions. Detailed Implementation

[0015] The present invention provides a pharmaceutical composition comprising, by weight: 1.5-2 parts of neem bark, 1-1.5 parts of Dryopteris crassirhizoma, 1-1.5 parts of Artemisia argyi, 0.8-1.2 parts of Artemisia annua, and 0.5-0.8 parts of Moringa root.

[0016] Unless otherwise specified, the present invention does not have any special limitations on the source of raw materials for the following technical solutions, and conventional commercially available products in the field can be used.

[0017] As an optional embodiment of the present invention, the pharmaceutical composition comprises 1.5 to 2 parts by weight of neem bark, which can be 1.5, 1.6, 1.7, 1.8, 1.9, or 2.0 parts. In this invention, the neem bark is preferably dried neem bark. In this invention, the neem bark, as the core anthelmintic component, contains azadirachtin, which has a strong paralyzing and killing effect on parasites such as Gyrodactylus and Trichodina, providing the basic anthelmintic efficacy for the formula. In this invention, the neem bark is obtained from the neem tree (Melia toosendan). Melia azedarach The root bark and bark of the tree.

[0018] As an optional embodiment of the present invention, based on the mass fraction of neem bark, the pharmaceutical composition includes 1-1.5 parts of Dryopteris crassirhizoma, which can be 1.0, 1.1, 1.2, 1.3, 1.4, or 1.5 parts. In the present invention, the Dryopteris crassirhizoma is preferably dried. In the present invention, the Dryopteris crassirhizoma and neem bark work synergistically to enhance the killing effect on Gyrodactylus, while broadening the anthelmintic spectrum and providing auxiliary killing effect on some intestinal parasites.

[0019] As an optional embodiment of the present invention, based on the mass fraction of neem bark, the pharmaceutical composition includes 1-1.5 parts of mugwort leaves, which can be 1.0, 1.1, 1.2, 1.3, 1.4, or 1.5 parts. In the present invention, the mugwort leaves are preferably dried mugwort leaves. In the present invention, the mugwort leaves target ectoparasites such as anchor worms and fish lice, and their active ingredients can penetrate to the gills of fish, inhibiting and killing worms such as *Dryopteris crassirhizoma* and *Cyclocarya paliurus* that parasitize the surface of the gills. In the present invention, the *Dryopteris crassirhizoma* is a plant of the Dryopteridaceae family (Dryopteris crassirhizoma). Dryopteris crassirhizoma The rootstock and petiole residues of ).

[0020] As an optional embodiment of the present invention, based on the mass fraction of neem bark, the pharmaceutical composition includes 0.8 to 1.2 parts of Artemisia annua, which can be 0.8, 0.9, 1.0, 1.1, or 1.2 parts. In the present invention, the Artemisia annua is preferably dried Artemisia annua. In the present invention, the Artemisia annua contains artemisinin, which is significantly effective against protozoan parasites such as Trichodina and Chilodonella, and when combined with neem bark and Dryopteris crassirhizoma, it further enhances the broad-spectrum anthelmintic effect. In the present invention, the Artemisia annua is Artemisia annua (also known as Artemisia annua), a plant of the Artemisia genus in the Asteraceae family. Artemisia annua The dried above-ground parts of ).

[0021] As an optional embodiment of the present invention, based on the mass fraction of neem bark, the pharmaceutical composition includes 0.5-0.8 parts of moringa root, which can be 0.5, 0.6, 0.7, or 0.8 parts. In the present invention, the moringa root is preferably dried moringa root. In the present invention, the moringa root contains active ingredients such as glucosinolates, alkaloids, and saponins, which can enhance the overall penetrability of the drug, help other drug components better penetrate into the depths of the fish gills, improve the expulsion and killing effect on deep-seated parasites, and its own active ingredients also have a certain parasite-repelling auxiliary effect. In the present invention, the moringa root is from the moringa tree (… Moringa oleifera The root of ).

[0022] As an optional embodiment of the present invention, the pharmaceutical composition further includes Tween-80. In this invention, based on the mass fraction of neem bark, the pharmaceutical composition includes 0.05 to 0.1 parts of Tween-80, which can be 0.05, 0.06, 0.07, 0.08, 0.09, or 0.1 parts. In this invention, Tween-80 acts as an emulsifier, enabling the extracts of various plant drugs to be uniformly dispersed in the water, preventing drug agglomeration, and ensuring that fish can uniformly contact the drug.

[0023] As an optional embodiment of the present invention, the pharmaceutical composition further includes ethanol. In this invention, the mass-to-volume ratio of neem bark to ethanol in the pharmaceutical composition can be (1.5~2) g:(5~8) mL, or it can be 1.8 g:6 mL, 1.5 g:5 mL, or 2.0 g:8 mL. In this invention, the ethanol mainly acts as a co-solvent, which can promote the dissolution of lipid-soluble active ingredients in the herbal medicine and improve the utilization rate of the active ingredients.

[0024] As an optional embodiment of the present invention, the composition of the pharmaceutical composition may be, by weight, 1.8 parts of neem bark, 1.2 parts of Dryopteris crassirhizoma, 1.2 parts of Artemisia argyi, 1.0 part of Artemisia annua and 0.6 parts of Moringa root.

[0025] As an optional embodiment of the present invention, the composition of the pharmaceutical composition may be, by weight, 1.5 parts of neem bark, 1.0 part of Dryopteris crassirhizoma, 1.0 part of Artemisia argyi, 0.8 parts of Artemisia annua and 0.5 parts of Moringa root.

[0026] As an optional embodiment of the present invention, the composition of the pharmaceutical composition may be, by weight, 2.0 parts of neem bark, 1.5 parts of Dryopteris crassirhizoma, 1.5 parts of Artemisia argyi, 1.2 parts of Artemisia annua, and 0.8 parts of Moringa root.

[0027] The pharmaceutical composition provided by this invention is a novel external formulation for expelling fish parasites, achieving broad-spectrum expulsion of various fish parasites such as Gyrodactylus, Trichodina, Chilodonella, Bodoworm, and Gyrodactylus, solving the problem of narrow spectrum expulsion of existing single drugs or simple drug combinations. The pharmaceutical composition enhances the targeting and penetration of the drug against parasites in the gills, improving the expulsion effect on parasites deep within the gills, thus solving the problem of incomplete expulsion of parasites in the gills by existing drugs. The pharmaceutical composition, through scientific formulation, leverages the synergistic effect of each drug component, reducing the concentration of single drugs while ensuring expulsion efficacy, minimizing stress on fish, and ensuring healthy fish growth. The pharmaceutical composition simplifies drug application, providing fish farmers with an efficient, convenient, and safe solution for fish parasite control.

[0028] This invention provides a method for preparing the pharmaceutical composition described above, comprising the following steps: The bark of neem tree, dryopteris crassirhizoma, mugwort leaves, artemisia annua and moringa root are crushed and mixed to obtain a raw material mixture; The raw material mixture is boiled to obtain an aqueous extract; The aqueous extract was concentrated to obtain a concentrated solution; The concentrate was mixed with ethanol and Tween-80 to obtain a pharmaceutical composition.

[0029] This invention involves pulverizing and mixing neem bark, Dryopteris crassirhizoma, Artemisia argyi, Artemisia annua, and Moringa root to obtain a raw material mixture. The pulverization method is not particularly limited; any conventional pulverization method in the art can be used. As an optional embodiment of this invention, the neem bark, Dryopteris crassirhizoma, Artemisia argyi, Artemisia annua, and Moringa root can be pulverized to 40-60 mesh, or to mesh sizes of 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 mesh. The mixing method is not particularly limited; any conventional mixing method in the art can be used.

[0030] After obtaining the raw material mixture, the present invention decocts the raw material mixture to obtain an aqueous extract. As an optional embodiment of the present invention, the decoction method of the raw material mixture includes: mixing the raw material mixture with water, performing a first decoction, and collecting the first decoction liquid and filter residue separately; mixing the filter residue with water, performing a second decoction, and collecting the second decoction liquid; and mixing the first and second decoction liquids to obtain the aqueous extract. In the present invention, the mass ratio of the raw material mixture to water can be 1:(10~15), or 1:10, 1:11, 1:12, 1:13, 1:14, or 1:15; the mass ratio of the filter residue to water can be 1:(8~10), or 1:9. The temperature of the first decoction can be 95~100℃, or 95, 96, 97, 98, 99, or 100℃; the decoction time can be 1.5~2 hours, or 1.5, 1.6, 1.7, 1.8, 1.9, or 2.0 hours; the temperature of the second decoction can be 95~100℃, or 95, 96, 97, 98, 99, or 100℃; the decoction time can be 1 hour. After decoction, an aqueous extract is obtained.

[0031] After obtaining the aqueous extract, the present invention concentrates the aqueous extract to obtain a concentrated solution. The present invention does not specifically limit the concentration method; any conventional concentration method in the art can be used. As an optional embodiment of the present invention, the concentration is preferably reduced to 1 / 3 of the original volume.

[0032] After obtaining the concentrate, the present invention mixes the concentrate with ethanol and Tween-80 to obtain a pharmaceutical composition. After obtaining the concentrate, the present invention cools the concentrate to room temperature and then mixes the resulting concentrate sequentially with ethanol and Tween-80. In the present invention, the room temperature can be 25°C. After each mixing, the present invention preferably stirs to obtain an emulsion. In the present invention, the stirring time after mixing the concentrate with ethanol can be 5 minutes; when mixing the concentrate-ethanol mixture with Tween-80, the stirring time can be 15-20 minutes, or 15, 16, 17, 18, 19, or 20 minutes. After stirring, the resulting emulsion can be directly used as a finished drug; the emulsion can also be referred to as a pharmaceutical composition.

[0033] This invention provides the application of the pharmaceutical composition described in the above-described technical solution or the pharmaceutical composition prepared by the above-described preparation method in the treatment of fish parasites. As an optional embodiment of this invention, the parasites include any one or more of the following: *Dactylogyrus*, *Trichodina*, *Chilodonella*, *Bodo*, and *Cyclophora*. As an optional embodiment of this invention, the fish can be any one or more of the following: carp, grass carp, and crucian carp.

[0034] This invention provides a method for eliminating parasites, comprising: applying the drug composition described in the above-described technical solution or the drug composition prepared by the preparation method described in the above-described technical solution to the water body for aquaculture of fish. As an optional embodiment of this invention, the application dosage may be 8-10 g / m³. 3 It can also be 8, 9 or 10 g / m 3 As an optional embodiment of the present invention, the water is preferably not changed within 24 hours after the drug composition is applied; the drug composition may be applied a second time depending on the killing of parasites after 72 hours. As an optional embodiment of the present invention, the application method may be spraying; when spraying, it may be slowly sprayed along the edge of the aquaculture pond to ensure uniform distribution of the drug.

[0035] The pharmaceutical composition provided by the present invention has the following advantages: 1. Broad spectrum of parasite control: Through the scientific combination of various herbal medicines, it can effectively kill a variety of common fish parasites such as Gyrodactylus, Trichodina, Chilodonella, Bodoworm, and Cupella, with a parasite control rate of over 95%, solving the problem of narrow parasite control spectrum of existing single drugs. 2. Strong targeting: With the enhanced penetrability of Moringa root, combined with the synergistic penetration effect of ingredients such as Artemisia argyi and Artemisia annua, the drug ingredients can effectively penetrate deep into the gills of fish, achieving a kill rate of over 92% for parasites in the gills, thus completely solving the problem of difficult-to-eliminate parasites in the gills of fish. 3. High safety: All plant raw materials are natural ingredients, and the dosage of each component in the embodiments of this invention does not exceed the concentration of a single anthelmintic. The synergistic effect produced by scientific compounding achieves high-efficiency anthelmintic treatment while avoiding the toxicity risks of using a single drug at high concentrations. The combination of Tween-80 and ethanol improves drug utilization. The stress response rate of fish after use is less than 5%, which will not affect the normal feeding and growth of fish. Compared with existing high-concentration single drugs, the safety is significantly improved. 4. Easy to operate: After the medicine is prepared, it can be directly sprayed throughout the pond without complicated steps. Farmers can operate it easily, which reduces the cost of aquaculture management.

[0036] To further illustrate the present invention, the technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0037] Example 1 A pharmaceutical composition, by weight, comprises: 1.5-2 parts dried neem bark, 1-1.5 parts dried Dryopteris crassirhizoma, 1-1.5 parts dried Artemisia argyi, 0.8-1.2 parts dried Artemisia annua, and 0.5-0.8 parts dried Moringa root.

[0038] Example 2 A pharmaceutical composition comprising: 1.5-2g of dried neem bark, 1-1.5g of dried Dryopteris crassirhizoma, 1-1.5g of dried Artemisia argyi, 0.8-1.2g of dried Artemisia annua, 0.5-0.8g of dried Moringa root, 0.05-0.1g of Tween-80, and 5-8mL of ethanol.

[0039] Example 3 A pharmaceutical composition comprising: 1.8g of dried neem bark, 1.2g of dried Dryopteris crassirhizoma, 1.2g of dried Artemisia argyi, 1g of dried Artemisia annua, 0.6g of dried Moringa root, 0.08g of Tween-80, and 6mL of ethanol.

[0040] The preparation method of the pharmaceutical composition includes the following steps: Raw material pretreatment: Grind the dried neem bark, Dryopteris crassirhizoma, Artemisia argyi, Artemisia annua, and Moringa root into 50 mesh and set aside; Extraction: Weigh the above-mentioned pulverized plant materials, mix them evenly, add 12 times the weight of water, heat to a gentle boil, and decoct at 95℃~100℃ for 1.8 hours. Filter and collect the first decoction. Add 9 times the weight of water to the residue, heat to a gentle boil, and decoct at 95℃~100℃ for 1 hour. Filter and collect the second decoction. Combine the two decoctions and concentrate them to 1 / 3 of their original volume at 60℃~70℃ under normal pressure to obtain a concentrated plant extract, referred to as the concentrate. Preparation: Cool the concentrate to 25°C, add 6 mL of ethanol, and stir for 5 minutes until the ethanol is completely dissolved; then add 0.08 g of Tween-80 and continue stirring for 18 minutes to form a uniform emulsion, which is the finished drug.

[0041] The dosage of this finished drug is for use in 1 cubic meter of water.

[0042] Example 4 A pharmaceutical composition comprising: 1.5g dried neem bark, 1.0g dried Dryopteris crassirhizoma, 1.0g dried Artemisia argyi, 0.8g dried Artemisia annua, 0.5g dried Moringa root, 0.05g Tween-80, and 5mL ethanol.

[0043] The preparation method of the pharmaceutical composition is the same as in Example 3.

[0044] Example 5 A pharmaceutical composition comprising: 2.0g of dried neem bark, 1.5g of dried Dryopteris crassirhizoma, 1.5g of dried Artemisia argyi, 1.2g of dried Artemisia annua, 0.8g of dried Moringa root, 0.1g of Tween-80, and 8mL of ethanol.

[0045] The preparation method of the pharmaceutical composition is the same as in Example 3.

[0046] Comparative Example 1 The dried bark of neem was used alone as a raw material for anthelmintic drugs, with a dosage of 2.0g per cubic meter of water. The method for preparing the finished drug from this raw material was as follows: the dried bark of neem was pulverized to 50 mesh, added to 12 times its volume of water and simmered for 1.8 hours, the decoction was filtered, the same amount of water was added, and the decoction was simmered a second time. The two filtrates were mixed and concentrated to 1 / 3 of the original volume.

[0047] Comparative Example 2 The dried product of *Dryopteris crassirhizoma* was used alone as a raw material for anthelmintic treatment, with a dosage of 1.5g per cubic meter of water. The method for preparing the finished drug from this raw material was the same as that for the finished drug prepared from *Melia azedarach* bark in Comparative Example 1.

[0048] Comparative Example 3 Dried Artemisia argyi was used alone as a raw material for the anthelmintic drug, with a dosage of 1.5g per cubic meter of water. The method for preparing the finished drug from this raw material was the same as that for the finished drug prepared from Melia azedarach bark in Comparative Example 1.

[0049] Comparative Example 4 Artemisia annua was used alone as the anthelmintic ingredient, with a dosage of 1.2g per cubic meter of water. The method for preparing the finished drug from this ingredient was the same as that for the neem bark finished drug in Comparative Example 1.

[0050] Comparative Example 5 Moringa root extract was used alone as the raw material for the anthelmintic drug, with a dosage of 0.8g per cubic meter of water. The method for preparing the finished drug from this raw material was the same as that for the neem bark finished drug in Comparative Example 1.

[0051] Comparative Example 6 A pharmaceutical composition, with the same composition and preparation method as in Example 3, except that it does not contain neem bark.

[0052] Comparative Example 7 A pharmaceutical composition, with the same composition and preparation method as in Example 3, except that it does not contain Moringa root.

[0053] Comparative Example 8 A pharmaceutical composition, with the same composition and preparation method as in Example 3, the only difference being that no mixing is performed during the preparation of the pharmaceutical composition (i.e., no ethanol and Tween-80 are added), and the concentrated solution obtained is directly used as the finished drug.

[0054] Application Example 1 The experiment was divided into 12 experimental groups, each using a fish farming pond with a water volume of approximately 100 cubic meters, with an area of ​​approximately 67-100 square meters and a water depth of 1-1.5 meters. The pond primarily housed carp, with a small number of grass carp and crucian carp also stocked to observe safety. The following test data and analysis are representative of carp. All fish in the experimental ponds exhibited typical parasitic infection symptoms such as rapid breathing and swollen gills. Preliminary testing revealed the presence of *Dactylogyrus*, *Trichodina*, *Chilodonella*, *Bodo*, and *Cyclophora* parasites on the gills and body surface, with a mixed infection rate exceeding 85%.

[0055] The experimental groups were the blank control group, the Chinese medicine composition group of Example 3, the Chinese medicine composition group of Example 4, the Chinese medicine composition group of Example 5, the drug group of Comparative Example 1, the drug group of Comparative Example 2, the drug group of Comparative Example 3, the drug group of Comparative Example 4, the drug group of Comparative Example 5, the drug group of Comparative Example 6, the drug group of Comparative Example 7, and the drug group of Comparative Example 8.

[0056] According to the ratio of the total amount of the above formula per cubic meter of water, evenly sprinkle the finished drugs prepared in Examples 3-5 and Comparative Examples 1-8 into the aquaculture pond. After sprinkling, maintain oxygenation and do not change the water for 24 hours. Specifically, do not change the water within 24 hours of drug application to ensure that the drug has sufficient time and concentration to exert its effect. After 24 hours, adopt a gradual water change method: each water change should be about 1 / 3 to 1 / 2 of the total pond water volume, with an interval of at least 4 hours between water changes, and complete the entire water change in 2-3 times within 24-48 hours. The new water should be aerated in advance to ensure that the water temperature and pH value are basically the same as the pond water to reduce stress on the fish. Oxygenation must be maintained during the water change process.

[0057] (1) Sampling: Each component of the medicine was evenly sprinkled into the pond according to the proportion of the total amount of the formula per cubic meter of water. Sampling and testing were carried out before medication, 24 hours after medication, and 72 hours after medication. Before medication, 10 carp with typical symptoms were randomly caught from each pond as preliminary samples. 24 hours and 72 hours after medication, 10 carp were randomly caught from each pond as efficacy evaluation samples.

[0058] (2) Testing: External parasites: Scrape the mucus from the fish's body surface, make a water-soaked slide, and observe and count it under a microscope (10×10 magnification).

[0059] Gill parasites: Cut off the second gill flap of each fish, place it on a glass slide, add a small amount of water, tear open the gill flap, cover with a coverslip, and observe and count under a microscope (10×10 magnification).

[0060] (3) Calculation: Parasite reduction rate (%) = [(Average number of parasites per fish before medication - Average number of parasites per fish at a certain time point after medication) / Average number of parasites per fish before medication] × 100%.

[0061] For example, the statement "24 hours after medication, the number of parasites in the fish gills decreased by 60%" is calculated using this formula.

[0062] Parasite eradication rate (%): refers to the percentage of fish tails in the test sample that did not show any parasites after a certain period of medication.

[0063] For example, "72 hours after medication, the parasite killing rate reached 98%" means that out of 10 fish tested, no individual parasites were detected in the gills and body surface of 9 to 10 fish.

[0064] Table 1. Results of drug application in Examples 3-5 and Comparative Examples 1-8 (carp)

[0065] Note: In Examples 3-5, the dosage of each single component did not exceed the single-component anthelmintic concentration in Comparative Examples 1-5 (e.g., 1.8 g / m³ of neem bark in Example 3). 3 <Comparative Example 1: 2.0 g / m 3 Dryopteris crassirhizoma 1.2g / m 3 <Comparative Example 2: 1.5 g / m 3 Artemisia argyi 1.2g / m 3 <Comparative Example 3: 1.5 g / m 3 Artemisia annua 1.0g / m 3 <Comparative Example 4: 1.2 g / m3 Moringa root 0.6g / m 3 <0.8 g / m² of Comparative Example 5 3 As can be seen, the present invention achieves significantly better anthelmintic effects (Table 2) and lower fish stress response (0% mortality rate in Table 1) through compound synergistic effects at lower or equivalent single component dosages, fully demonstrating the technical effects of the drug composition provided by the present invention in terms of low concentration, high efficiency, and low stress.

[0066] Table 2. Kill rate of different parasites at 72h

[0067] In Example 3, 24 hours after administration of the drug composition, 10 carp were randomly caught for testing. It was found that the parasites on the body surface had basically disappeared, and the number of parasites in the gills was reduced by 60%. 72 hours after administration of the drug composition of Example 3, 10 carp were randomly caught again for testing. No parasites were detected in the gills or on the body surface, and the parasite eradication rate reached 98%. The carp's breathing returned to normal, their appetite was significantly enhanced, and no fish deaths or obvious stress reactions were observed.

[0068] In Example 4, the drug composition was tested on randomly caught carp 24 hours after administration, and the parasite eradication rate was found to be 55%. After 72 hours of administration, another 10 randomly caught carp were tested, and the parasite eradication rate reached 96%. This indicates that the formulation has a sustained and significant efficacy.

[0069] The drug composition of Example 5, when tested on randomly caught carp 24 hours after administration, showed a parasite eradication rate of 58%. After 72 hours of administration, another 10 randomly caught carp were tested, and the parasite eradication rate reached 97%. This demonstrates an excellent and thorough parasite-repelling effect.

[0070] One week after the drug compositions in Examples 3-5 were administered, a follow-up visit showed that the carp in the breeding pond were growing well and no recurrence of parasites was observed.

[0071] In contrast, the dosage of each single component in the embodiments of the present invention does not exceed the dosage of the single medicinal material used in comparative examples 1 to 5, and is even lower. However, under the synergistic effect of the compound, the anthelmintic effect is significantly improved and the stress response of fish is significantly reduced, which fully demonstrates that the present invention achieves the technical effects of low concentration, high efficiency and low stress through scientific formulation.

[0072] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A pharmaceutical composition, characterized by, By weight, it includes: 1.5-2 parts of neem bark, 1-1.5 parts of Dryopteris crassirhizoma, 1-1.5 parts of Artemisia argyi, 0.8-1.2 parts of Artemisia annua, and 0.5-0.8 parts of Moringa root.

2. The pharmaceutical composition according to claim 1, wherein By weight, it includes: 1.8 parts of neem bark, 1.2 parts of Dryopteris crassirhizoma, 1.2 parts of Artemisia argyi, 1.0 part of Artemisia annua, and 0.6 parts of Moringa root.

3. The pharmaceutical composition according to claim 1 or 2, characterized in that, The pharmaceutical composition also includes Tween-80 and ethanol.

4. A method for preparing the pharmaceutical composition according to any one of claims 1 to 3, characterized in that, Includes the following steps: The bark of neem tree, dryopteris crassirhizoma, mugwort leaves, artemisia annua and moringa root are crushed and mixed to obtain a raw material mixture; The raw material mixture is boiled to obtain an aqueous extract; The aqueous extract was concentrated to obtain a concentrated solution; The concentrate was mixed with ethanol and Tween-80 to obtain a pharmaceutical composition.

5. The preparation method according to claim 4, characterized in that, Methods for decocting the raw material mixture include: Mix the raw material mixture with water and boil it for the first time. Collect the first decoction and the residue separately. Mix the filter residue with water and boil it a second time, then collect the second decoction. The first decoction and the second decoction are mixed to obtain an aqueous extract.

6. The preparation method according to claim 5, characterized in that, The mass ratio of the raw material mixture to water is 1:(10~15); the mass ratio of the filter residue to water is 1:(8~10).

7. The preparation method according to claim 5, characterized in that, The first decoction time is 1.5 to 2 hours; the second decoction time is 1 hour.

8. The use of the pharmaceutical composition according to any one of claims 1 to 3 or the pharmaceutical composition prepared by the preparation method according to any one of claims 4 to 7 in the treatment of fish parasites.

9. The application according to claim 8, characterized in that, The fish species include any one or more of carp, grass carp, and crucian carp; the parasites include any one or more of dactyloides, trichodina, chilodonella, bodoworm, and cupella.

10. A method for removing parasites from fish, characterized in that, include: The pharmaceutical composition according to any one of claims 1 to 3 or the pharmaceutical composition prepared by any one of claims 4 to 7 shall be applied to the water body for aquaculture of fish.

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